Chloroplast-based expression system is promising for the hyper-expression of plant-derived recombinant therapeutic proteins and vaccines. To verify the feasibility of obtaining high-level expression of the SARS subunit vaccine and to provide a suitable plant-derived vaccine production platform against the severe acute respiratory syndrome coronavirus (SARS-CoV), a 193-amino acid fragment of SARS CoV spike protein receptor-binding domain (RBD), fused with the peptide vector cholera toxin B subunit (CTB), was expressed in tobacco chloroplasts. Codon-optimized CTB-RBD sequence was integrated into the chloroplast genome and homoplasmy was obtained, as confirmed by PCR and Southern blot analysis. Western blot showed expression of the recombinant fusion protein mostly in soluble monomeric form. Quantification of the recombinant fusion protein CTB-RBD was conducted by ELISA analysis from the transplastomic leaves at different developmental stages, attachment positions and time points in a day and the different expression levels of the CTB-RBD were observed with the highest expression of 10.2% total soluble protein obtained from mature transplastomic leaves. Taken together, our results demonstrate the feasibility of highly expressing SARS subunit vaccine RBD, indicating its potential in subsequent development of a plant-derived recombinant subunit vaccine and reagents production for antibody detection in SARS serological tests.
Chloroplasts ; metabolism ; Cholera Toxin ; Protein Interaction Domains and Motifs ; Recombinant Fusion Proteins ; biosynthesis ; SARS Virus ; Spike Glycoprotein, Coronavirus ; biosynthesis ; Tobacco ; metabolism ; Vaccines, Subunit ; biosynthesis

Objective:To analyze the effects of different anesthesia depths on serum S100β protein level and postoperative cognitive function in patients undergoing minimally invasive McKeown esophagectomy with bispectral index (BIS) monitoring.Methods:A total of 120 patients who received minimally invasive McKeown esophagectomy in the Second Hospital of Shanxi Medical University from April 2018 to April 2019 were selected, and they were divided into light anesthesia group (L group, 40 cases, BIS 46-60), deep anesthesia group (D group, 40 cases, BIS 35-45) and control group (C group, 40 cases, no BIS monitoring) by using the random number table method. Elbow venous blood was taken from each group to detect serum S100β protein levels before induction (T 0), 10 minutes after extubation (T 3), the first day after surgery (T 4), and the third day after surgery (T 5). The mini-mental state examination scale (MMSE) score and the monterey cognitive assessment scale (MoCA) score were performed before surgery and on day 1, 3, and 7 after surgery to count the incidence of postoperative cognitive dysfunction(POCD). Results:There was no statistical difference in serum S100β protein levels between the three groups at T 0 (F = 0.083, P = 0.920). The level of serum S100β protein in D group [(1.08±0.05) μg/L] was significantly higher than that in C group and L group [(0.98±0.10) μg/L and (0.84±0.09) μg/L] at T 3, and the level of serum S100β protein in L group was lower than that in C group, the differences between the three groups were statistically significant (P < 0.05). There was no statistical difference in the incidence of POCD among the three groups on day 7 after surgery (χ 2 = 2.914, P = 0.233). The incidence rates of POCD in D group on day 1 and 3 after surgery (57.1% and 37.1%) were significantly higher than those in C group (41.7% and 38.9%) and L group (20.0% and 14.3%), and the incidence of POCD in L group were lower than those in C group, the differences among the three groups were statistically significant (χ 2 = 10.187, P = 0.006; χ 2 = 6.296, P = 0.043). Conclusions:For patients undergoing minimally invasive McKeown esophagectomy, intraoperative BIS monitoring maintains a light anesthetic state, which can effectively reduce serum S100β protein level and POCD. The mechanism may be related to reducing serum S100β protein level and improving brain damage.

Objective:To investigate the effects of propofol combined with sevoflurane intravenous-inhalational anesthesia and propofol intravenous anesthesia on implicit memory, explicit memory and stress response in patients undergoing gynecological cancer surgery.Methods:A total of 48 patients undergoing gynecologic cancer surgery in the Second Hospital of Shanxi Medical University from August 2018 to May 2019 were selected. Random number table was used to divide patients into propofol intravenous anesthesia group (group A) and propofol combined with sevoflurane intravenous-inhalational anesthesia group (group B), 24 cases in each group. During the operation, the patients in group A were given continuous intravenous anesthesia with propofol and the patients in group B were given continuous intravenous anesthesia with propofol combined with sevoflurane. Minimal alveolar concentration (MAC) of sevoflurane was 0.5, and midazolam was not utilized throughout the whole anesthesia for both groups. The bispectral index (BIS) value of the two groups ranged from 45 to 55 during the operation. The concentration of cortisol, adrenocorticotropic hormone (ACTH) and prolactin(PRL) in elbow venous blood was measured before anesthesia induction (T 1) and 10 minutes after intraoperative auditory recording (T 2). Implicit and explicit memory scores were measured 12-24 h after operation. Results:Neither group produced explicit memory compared with 0 (group A: 0.012±0.007, t = 1.554, P > 0.05; group B: 0.016±0.002, t = 1.942, P > 0.05), and there was no significant difference in explicit memory score between the two groups ( t = -0.417, P > 0.05). Both groups produced implicit memory (group A: 0.089±0.050, t = 8.726, P < 0.05; group B: 0.189±0.060, t = 15.415, P < 0.05), and implicit memory score was higher in group B ( t = -6.215, P < 0.05). The level of cortisol, ACTH and PRL at time T 1 was not significantly different between the two groups (all P > 0.05), and the level of cortisol, ACTH and PRL in group B was higher than that in group A at time T 2 [(276±35) μg/L vs. (96±33) μg/L; (228±42.3) pg/ml vs. (14.1±1.7) pg/ml; (4 208±213) mU/ml vs. (3 805±196) mU/ml; t value was 18.634, 34.879, 12.605, all P < 0.05]. Conclusion:Propofol intravenous anesthesia can better inhibit the production of implicit memory and intraoperative stress response compared with propofol combined with sevoflurane intravenous-inhalational anesthesia.

Objective: To study the effects of different anesthesia depths on stress response during single-lung ventilation in patients with thoracoscopic lobectomy. Methods: Sixty patients selected for elective thoracoscopic lobectomy in the Second Hospital of Shanxi Medical University from September 2018 to May 2019 were randomly divided into three groups according to the digital random table method, with 20 patients in each group. Group A maintained deep anesthesia with the bispectral index (BIS) 36-45, group B maintained moderate anesthesia with BIS 46-55, and group C did not undergo BIS monitoring. The changes of heart rate, mean arterial pressure (MAP), stress indexes cortisol and blood glucose before anesthesia induction (T₀), immediately after one-lung ventilation (T₁), 60 min after one-lung ventilation (T₂) and immediately after skin suture (T₃) in the three groups were compared. Results: The concentration of blood glucose in group A at T₁, T₂ and T₃ was (5.28±0.49) mmol/L, (5.34±0.49) mmol/L and (5.40±0.47) mmol/L, and the cortisol was (142.75±31.45) ng/ml, (181.36±19.62) ng/ml and (153.81±33.92) ng/ml; the blood glucose in group B was (5.63±0.35) mmol/L, (6.06±0.19) mmol/L and (5.79±0.44) mmol/L, and the cortisol was (168.45±31.16) ng/ml, (171.09±25.28) ng/ml and (159.39±18.77) ng/ml; the blood glucose in group C was (6.35±0.56) mmol/L, (7.04±0.26) mmol/L and (6.17±0.54) mmol/L, and the cortisol was (191.13±46.00) ng/ml, (283.25±30.07) ng/ml and (183.01±19.71) ng/ml, respectively. The blood glucose and cortisol levels in group C at T₁, T₂ and T₃ were higher than those in group A and group B (all P < 0.05). The MAP in group A at T₁, T₂ and T₃ were (69±5) mmHg (1 mmHg= 0.133 kPa), (67±6) mmHg and (75±7) mmHg, respectively, and group B was (80±8) mmHg, (79±4) mmHg and (84±9) mmHg, the differences between the two groups were statistically significant (all P < 0.05). There was significant difference in cortisol between group A and group B at T₁ (P < 0.05). The heart rate and MAP at T₁, T₂ and T₃ in group A and group C were significantly different from those at T₀ (all P < 0.05). The heart rate and MAP at T₁ and T₂ in groups B were significantly different from those at T₀ (all P < 0.05). Conclusion BIS anesthesia depth monitoring should be performed during single-lung ventilation in thoracic surgery, and BIS should be maintained at 46-55, which can not only inhibit the stress response but also have a slight effect on hemodynamics.