In order to establish an infectious clone for CDV-3, a commercial vaccine strain of canine distemper virus for mink, to provide reference for the studies of pathogenesis and novel vaccine development of CDV. Thirteen pairs of primers were used to amplify the full-length genome of CDV-3 strain. Five long fragments were obtained based on single restriction site analysis of the whole genome of CDV-3 by RT-PCR. Five fragments were successively inserted into the multiple clone sites in the modified eukaryotic vector of pcDNA3.2 by restriction enzymes and splicing. Meanwhile, the hammerhead ribozyme and hepatitis delta virus ribozyme sequences were added to the beginning of F1 fragment and the ending of F5 fragment, respectively. Then, the full-length cDNA recombinant plasmid of CDV-3 was obtained and named as pcDNA3.2-CDV-3. In addition, three helper plasmids, expressing the N protein, P protein and L protein of the CDV-3 strain respectively, were constructed. The 293T cells were transfected with the full-length cDNA recombinant plasmid and three helper plasmids by Lipofectamine™ 2000. At 3 days post transfection, the supernatant was added to the monolayer of Vero cells to observe the typical syncytium of CDV. Indirect immunofluorescence and artificial label identification of recombinant virus rCDV-3 were conducted after the occurrence of lesions. Finally, the growth characteristics of wtCDV-3 and rCDV-3 were compared after passaging of rCDV-3. The identification of the full-length cDNA recombinant plasmid and three helper plasmids by restriction enzyme digestion and sequencing were consistent with expected. The Vero cells infected with the recombinant rCDV-3 showed typical syncytic. The identification of indirect immunofluorescence and labeled marker, and observation under electron microscope proved that the rCDV-3 was indeed rescued from the recombinant plasmid of pcDNA3.2-CDV-3. In comparison of the virus titers of wtCDV-3, rCDV-3 replicated massively and rapidly and reached the maximize virus titer of 10⁷·⁶⁶⁷ TCID₅₀/mL within 36 h post infection (p.i.) in Vero cells, while wtCDV-3 grew gradually to 10⁶·⁶⁶⁷ TCID₅₀/mL at 72 h p.i. in Vero cells. This reverse genetic system of CDV-3 strain has been established successfully, to provide reference for the studies of pathogenesis and novel vaccine development of CDV.
Animals ; Chlorocebus aethiops ; Clone Cells ; DNA, Complementary ; Distemper Virus, Canine/genetics* ; Plasmids/genetics* ; Vero Cells

Objective:To explore the effects of standardized enteral nutrition process in critically ill patients.Methods:We retrieved PubMed, EMBASE, MEDLINE, WanFang Data, China National Knowledge Infrastructure (CNKI) and China Biology Medicine (CBM) by computer to collect literatures on effects of standardized enteral nutrition process in critically ill patients from the establishment of database to May 31, 2019. RevMan 5.3 was used to statistical analysis.Results:Finally, 11 articles were included. Meta-analysis results showed that compared with conventional care, the standardized enteral nutrition process could increase the calorie compliance rate of critical ill patients on the seventh day [ OR=8.18, 95% CI (4.91, 11.45) , P<0.01], and reduce the incidence of feeding intolerance symptoms [ OR=0.36, 95% CI (0.23, 0.57) , P<0.01]. However, the standardized enteral nutrition process did not show obvious advantages in shortening mechanical ventilation, Intensive Care Unit (ICU) hospitalization time and reducing mortality, combined effects, the combined effect was not statistically significant ( P>0.05) . Conclusions:The standardized enteral nutrition process could effectively increase the calorie compliance rate of critical ill patients on the seventh day and reduce the incidence of feeding intolerance symptoms; however, it cannot significantly shorten the time of mechanical ventilation, ICU hospitalization time and reduce the mortality. Further research is needed for verification.

Objective:To explore the effects of individualized exercise prescription in critically ill patients during enteral nutrition.Methods:From January to August 2018, we selected 120 critically ill patients admitted to the General Surgery Intensive Care Unit (ICU) of General Hospital of Eastern Theater Command. According to admission time, patients were divided into control group and observation group, with 60 cases in each group. Control group used the conventional chest physical therapy, exercise program for passive and active functional exercise of limbs. Observation group implemented the individualized exercise prescription to implement functional exercise orders. After daily rounds, doctors and nurses assessed the condition of the day by combining patient's basic condition and prescribed the individualized exercise prescription, including functional exercise items, requirements, dosages and frequency, performed by a full-time nurse. We compared the muscle strength of patients between two groups with the MRC scores, and observed the intolerance, number of interruptions of enteral nutrition feeding and the ICU hospitalization time during enteral nutrition between two groups.Results:After 168 hours and 366 hours, the differences of MRC scores between two groups were significant ( P<0.05) . After 14 days of intervention, the serum total protein, hemoglobin and body mass index (BMI) of observation group were higher than those of control group also with statistical differences ( P<0.05) . The number of cases of abdominal distension, gastric retention, the total number of intolerance and the number of feeding interruptions during enteral nutrition in the observation group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . The ICU hospitalization time of patients of observation group and control group were (25.28±5.42) d and (27.94±5.65) d respectively also with a statistical difference ( P<0.05) . Conclusions:The individualized exercise prescription functional exercise intervention strategies is beneficial for critically ill patients to improve muscle strength, nutritional indicators, avoid rapid decline in BMI, reduce feeding intolerance and the number of feeding interruptions, ensure continuous implementation of enteral nutrition, and shorten patients' ICU hospitalization time so as to promote accelerated recovery of patients.

Critical ill children are generally at risk of malnutrition.Malnutrition can reduce immunity and cause a series of problems such as growth and development stagnation, increased infections and aggravation of illness. Early nutritional support is an important part of treatment and an important guarantee for successful treatment of children. This article reviews the status of early enteral nutrition, intolerance-related symptoms, tolerance monitoring and process management in critical ill children.

Objective:To explore the influences of different lying positions on vital signs and comforts in patients with tracheal intubation removed after intestinal fistula surgery.Methods:From October 2018 to June 2019, convenience sampling was used to select 150 patients with intestinal fistula surgery after general anesthesia in Intensive Care Unit of the General Hospital of Eastern Theater Command were selected as the research object. After waiting for the patient to wake up from anesthesia, the tracheal intubation was removed, and the lying position was changed after normal spontaneous breathing through the nose. Patients were randomly divided into 5 groups, 30 cases in each group. Group A was in the supine position; group B was in the lying position with the head of the bed raised by 15°; group C was in the lying position with the head of the bed raised by 30°; group D was in the lying position with the head of the bed raised by 45°; group E was in the lying position with the head of the bed raised by 15° to 45 °. At the beginning, the head of the bed was shaken to 15 °; and after two hours, the head of the bed was shaken to 30 °, and after another two hours, the head of the bed was shaken to 45 °. After 5 groups of patients fixed the angle of the lying position, when the patient complained of discomfort or the body position changed spontaneously, the patient changed the lying position. The duration of the lying position, uncomfortable symptoms and pain degree of abdominal incision were compared among 5 groups.Results:Patients with tracheal intubation removed after intestinal fistula surgery had a longer persistence in the 15° and 30° lying positions within 6 hours, respectively (72.27±12.41) min and (69.37±9.10) min. There were statistically significant differences in the persistence time of 5 different lying positions ( P<0.01) . The number of patients in group B and C with waist pain was less, and there were more patients with painless incisions, and there were statistically significant differences among 5 groups ( P<0.05) . Conclusions:Among patients with tracheal intubation removed and normal spontaneous breathing through the nose after intestinal fistula surgery, the 15° to 30° lying position within 6 hours after the operation can prolong the patient's adherence to the lying position, reduce the incidence of abdominal incision pain and waist pain, and reduce the probability of dizziness, nausea and vomiting.

Intra-abdominal hypertension can reduce the perfusion of the mesenteric artery, block venous return and reduce gastrointestinal function due to intestinal edema. And the delay of intestinal emptying can easily cause intestinal edema, which further increases intra-abdominal pressure and forms a vicious circle. Nutritional support plays an important role in treatment of severe patients with abdominal hypertension, as people have a deeper understanding of the importance of central organ of the intestinal stress response and the function of the intestinal barrier. This paper summarizes the implementation of enteral nutrition management for severe patients with abdominal hypertension, implements the standardized process of enteral nutrition implementation, evaluates nutritional risks, calculates target calorie intake and dynamically monitors intra-abdominal pressure, so as to ensure safety and effectiveness of patients in the process of nutritional support and to improve nutritional indexes and prognosis.

Objective:To explore the influences of abdominal pressure monitoring in different positions on patients with abdominal hypertension.Methods:From January to June 2019, convenience sampling was used to select 100 critical ill patients with abdominal hypertension who were admitted to the Intensive Care Unit of General Surgery of the General Hospital of Eastern Theater Command as the research object. The abdominal pressure was monitored by indirectly measuring the bladder pressure. We measured the abdominal pressure when patients were in stable condition and in 30° lying position, 45° lying position as well as the supine position respectively, recorded the single measurement data of the patient's abdominal pressure in different positions, and performed another position measurement rest for 15 minutes after changing the position.Results:The abdominal pressures measured in 100 critical ill patients with abdominal hypertension in the supine position, 30° lying position and 45° lying position were (19.18±3.95) , (23.40±3.87) and (28.17±3.60) mmHg (1 mmHg=0.133kPa) respectively, and the difference was statistically significant ( P<0.01) . The abdominal pressure monitored in the supine position was lower than those in the 30° lying position and 45° lying position, the difference was statistically significant (supine position vs. 30° lying position, P<0.01; supine position vs. 45° lying position, P<0.01) . The abdominal pressure monitored in the 30 ° lying position and 45 ° lying position was higher than that in the supine position by one to two levels. With the increase of the bed head elevation angle, the abdominal pressure value is getting higher and higher, regardless of the level of abdominal pressure in critical ill patients with abdominal hypertension. Conclusions:The abdominal pressure monitoring in actual positions can reflect the true situation of the patient well, and can provide a relatively true and reliable monitoring data for clinical nursing decision-making.

Objective To analyze the effects of structured fat emulsion and medium/long chain fat emulsion on blood lipids,immune cells and acute inflammation after hepatectomy for hepatocellular carcinoma.Methods Total of 60 patients with hepatocellular carcinoma who underwent hepatectomy in Henan People's Hospital (Zhengzhou University People's Hospital) from January 2013 to March 2017 were divided into experimental group (using structured fat emulsion) and control group (using medium/long chain fat emulsion),30 cases in each group.Triglyceride (TG),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),total cholesterol (TC),T lymphocyte level,fibrinogen (FIB),C-reactive protein (CRP),prostaglandin E2 (PGE2) were detected before and 2,4,6 and 8 days after infusion.Results There were no significant differences in LDL-C,HDL-C,TG and TC between the two groups before infusion (P＞0.05).On the 2nd day of parenteral nutrition infusion,the level of blood lipids in both groups was higher than experimental group before infusion;on the 4th,6th and 8th day of infusion,LDL-C,HDL-C,TG and TC in the control group were higher than those in the experimental group (P＜0.05).After parenteral nutrition infusion,the levels of CD3+,CD4+,CD8+,CD4+/CD8+ in both groups were higher than experimental group before infusion,and the experimental group was higher than the control group,the differences were statistically significant (P＜0.05).Compared with before infusion,level of FIB,CRP and PGE2 began to increase on the 2nd day of infusion,and the differences were statistically significant (P＜0.05).On the 2nd,4th,6th and 8th day,CRP in the control group was higher than experimental group.And resepeatively (19.12±5.84) mg/ml vs.(13.76±2.36) mg/ml,(31.67±8.68) mg/ml vs.(17.21±2.66) mg/ml,(22.15±8.33) mg/ml vs.(12.48±0.63) mg/ml,(9.65±4.66) mg/ml vs.(7.52±0.99) mg/ml,and PGE2 were also higher than that in the experimental group (P＜0.05).Conclusion Structured fat emulsion is superior to medium/long chain fat emulsion in improving blood lipid,immune cells and inflammatory reaction in patients after hepatectomy.

Objective? To summarize the key content of nursing of stepped drainage new model for severe acute pancreatitis (SAP) patients with necrosis and infection. Methods? From January 2014 to December 2016, we selected 417 SAP patients with infected pancreatic necrosis (IPN) in General Surgery Intensive Care Unit at Nanjing General Hospital, People's Liberation Army. The "four-step" stepped drainage model was adopted during treatment including four stages, percutaneous catheter drainage, continuous negative pressure irrigation drainage, endoscopic drainage and laparotomy necrotic tissue debridement drainage. All stages of nursing had something in common and different emphasis points. Nursing care paid attention to strengthening the nursing for abdominal puncture catheter, abdominal double pipe, endoscopic operation cooperation and postoperative nursing of laparotomy. Results? Among 417 SAP patients with IPN, a total of 413 patients were with percutaneous catheter drainage and single patient was with one to seven drainage tubes, a total of 2 252 tubes; 315 patients were with abdominal double pipe irrigation drainage and single patient was with two to seven double pipes, a total of 1 386 double pipes; 186 patients were with endoscopic drainage and single patient was with two to five times of endoscopic necrotic tissue debridement, a total of 725 times; 91 patients were with laparotomy and single patient was with laparotomy one to three times. During nursing care, totals of 13 percutaneous drainage catheters and abdominal double pipes slipped off; 109 cases of poor drainage happened to double pipe drainage with 56 cases of obstructed drainage;there were 30 cases of folding, 7 cases of hemorrhage and 16 cases of shifting. Those problems were all found and handled timely with no consequence caused by poor drainage and improper nursing care. Conclusions? Among SAP patients with IPN, keys to guarantee high quality of nursing included mastering nursing emphasis of all stages, taking targeted nursing, keeping abdominal puncture catheter unobstructed, abdominal double pipes effectively sucking, cooperating on endoscopic necrotic tissue debridement and strengthening postoperative monitoring and wound management.

Objective? To explore the current status of feeding interruption of early enteral nutrition among critical patients and its related factors. Methods? From June 2017 to March 2018, we selected critical patients with early enteral nutrition at a ClassⅢ Grade A hospital in Nanjing by convenience sampling. We recorded the enteral nutrition intake of patients within one week. All of the patients were divided into three groups based on the total time of feeding interruption including group A (＜4 hours), group B (4 to 24 hours) and group C (＞24 hours). The general information, calorie and prognostic factors of patients in three groups were compared and the related factors of feeding interruption time were analyzed with the multiple linear regression. Results? A total of 198 patients were enrolled including 108 of them in group A, 56 in group B and 34 in group C. There was a statistical difference in the completion rate of actual calorie in three groups (P＜ 0.05). Multiple linear regression analysis showed that the feeding interruption time of enteral nutrition were positively correlated with the disease severity, hospitalization expense and target calorie (P＜0.05). Conclusions? The incidence of feeding interruption is high in critical patients with enteral nutrition. The longer the interruption time is, the higher the risk of caloric inadequacy. At present, there is no standardized feeding strategy which needs to explore the best intervention by clinical researches so as to solve the problem of feeding interruption among critical patients.