According to the codon bias of Pichia pastoris, the mature insect neurotoxin gene LqhIT2 was synthesized based on its amino acid sequence and was cloned to vector of PET-30a (+) and pPIC9K respectively. The fusion protein expressed in Escherichia. coli was induced with IPTG and purified with Ni-NTA His Bind Column. The purified fusion protein was used to immunize BALB/c mice, and antiserum obtained was highly specific with the titer of over 1:128 000. Using the antiserum, high-level expression transformants of P. pastoris were screened by dot blotting. The highest expression of recombinant LqhIT2 was about 9 mg/L in baffled flasks. The fusion protein of LqhIT2 expressed in E. coli was not toxic to locust, but the recombinant LqhIT2 expressed in P. pastoris had insecticidal activity against locust through injection.
Animals ; Escherichia coli ; genetics ; metabolism ; Immune Sera ; biosynthesis ; Immunization ; Mice ; Mice, Inbred BALB C ; Neurotoxins ; biosynthesis ; genetics ; immunology ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Scorpion Venoms ; biosynthesis ; genetics ; immunology

AIM: To investigate the changes of bcl-2, bax expression and neuron apoptosis of cerebral cortex in lymphostatic encephalopathy of rats. METHODS: The model of lymphostatic encephalopathy was established by occluding and removing both the shallow and deep cervical lymph nodes in rats. The animals were sacrificed at 1, 2, 3, 5, 7 and 14 days after operation. HE staining was used to observe the structure of brain tissues and TUNEL staining was used to detect in situ cell apoptosis. The expressions of bcl-2 and bax were examined by RT-PCR. RESULTS: Cerebroedema appeared at the second day and was the most serious at the 5th day after blockage of cervical lymphatics. The number of TUNEL positive cells and the expression of bax began to increase at the 2nd day, reached a peak at the 5th day and dropped to control level at the 14th day. The expression of bcl-2 began to increase at the 1st day, reached a peak at the 5th day and dropped to control level at the 7th day. The increasing extent of bax was higher than that of bcl-2. CONCLUSION: The blockage of cervical lymphatics can lead to lymp[JP2]hostatic encephalopathy. Apoptosis is the main form of neuron death in the cortex and has relation to the increasing expression of bcl-2 and bax. [JP]

Objective To explore the best time for indwelling gastric tube after tracheal stenosis in infants with congenital heart disease, so as to shorten the indwelling time and relieve the pain of children with tracheal tube. Methods A total of 50 infants with congenital heart disease combined with tracheal stenosis were divided into observation group (26 cases) and control group (24 cases) by the admission order. The observation group accepted swallowing function evaluation combined with physiological indicators to determine the timing of removal of indwelling gastric tube while the control group just conventionally evaluating swallowing function. Results The ICU retention time and total hospitalization days was (4.35 ± 0.94), (23.15 ± 4.92) d in the observation group, and (6.27 ± 1.42), (27.42 ± 6.43) d in the control group, and the difference between the two groups was statistically significant (Z=5.589, 2.621, P<0.05). The extubation time was (2.85 ± 0.23), (4.50 ± 0.27) days in the observation group and the control group, and the difference was statistically significant (χ2= 15.595, P<0.01). Conclusions Deglutition function assessment combined with physiological indicators for evaluating the time of retention of gastric tube after tracheal stenosis in infants with congenital heart disease could be more effective to reduce the occurrence of secondary intubation and postoperative complications,shorten the course of the disease, improve infants comfort level,promote postoperative recovery.

Background Occupational stress has been shown to be an important factor affecting the mental health of workers. The role of affective commitment to the organization and overcommitment to work cannot be ignored. However, there is a lack of research on this topic in China. Objective To explore a potential mediating effect of affective commitment on how occupational stress affects the mental health of medical staff and a potential moderating effect of overcommitment on the mediating effect of affective commitment. Methods A total of 1372 health care workers in a tertiary Grade A hospital in Lanzhou City were selected as study subjects for a cross-sectional survey. The occupational stress, emotional commitment, and psychological distress of the subjects were evaluated by the Effort-Reward Imbalance Scale, Affective Commitment Scale, and Kessler 10 Scale. SPSS 26.0 was used for correlation analysis, mediation analysis, and moderated mediation analysis. Common method bias wasevaluated by Harman one-factor test. Results A total of 1372 questionnaires were distributed, of which 1277 valid questionnaires were returned, with a valid recovery rate of 93.08%. The mean occupational stress score was 1.14±0.23, the mean overcommitment score was 20.26±3.21, the mean affective commitment score was 20.25±3.34, and the mean psychological distress score was 26.26±7.90. The Spearman correlation analysis results showed that occupational stress among medical staff was positively correlated with overcommitment and psychological distress (r=0.153, 0.410, P＜0.01) and negatively correlated with affective commitment (r=−0.341, P＜0.01); overcommitment was negatively related to affective commitment and positively related to psychological distress (r=−0.107, 0.312, P＜0.01); affective commitment was negatively related to psychological distress (r=−0.464, P＜0.01). The positive effect of occupational stress on psychological distress of medical staff was significant (b=0.41, t=15.42, P<0.001); affective commitment presented a partial mediating effect on the relationship between occupational stress and psychological distress (effect value=0.13), accounting for 31.71% of the total effect; overcommitment moderated the process of occupational stress-affective commitment-psychological distress (P＜0.01). Conclusion Affective commitment of medical staff has a partial mediating effect on the relationship between occupational stress and psychological distress, and overcommitment plays a significant role in moderating the process of occupational stress-affective commitment-psychological distress.

OBJECTIVETo observe the effects of increased-intensity conditioning regimen with FBCA (Fludarabine, Busulfan, Cyclophosphamide, and Antithymocyte globulin) for allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acquired severe aplastic anemia (SAA).

METHODSFrom January 2000 to June 2011, twenty-two patients (male 12, female 10) with SAA underwent allo-HSCT with FBCA conditioning regimen which consisted of fludarabine (30 mg·m⁻²·d⁻¹×5 d), busulfan (3 mg/kg×2 d), cyclophosphamide (60 mg·kg⁻¹·d⁻¹×2 d) and ATG (2.5 mg·kg⁻¹·d⁻¹×5 d). GVHD prophylaxis was performed by cyclosporine and short-term course methotrexate. Nine patients received mobilized peripheral blood stem cells transplantation and 13 patients underwent mobilized peripheral blood combined with bone marrow stem cells. Fourteen cases were human leukocyte antigen (HLA)-matched related donors, while the other 8 cases were HLA-haploidentical transplantation. Engraftment was documented by short tandem repeats with polymerase chain reaction (STR-PCR) on approximately day + 30, + 90, + 180, + 1 year and + 2 year, respectively. Long-term survival and transplantation-related complications were analyzed.

RESULTSAll patients obtained prompt and sustained hematopoietic reconstitution. Median time for neutrophil and PLT engraftment was 15 (range: 11-22) days and 16 (range: 12-27) days, respectively. All patients were full donor chimerism identified by STR-PCR. 2 of the total 22 cases (9.1%) had grade I-III acute GVHD and 3 (15.8%) was chronic GVHD. Three patients (13.6%) died of transplantation related mortality and the other 19 cases were disease-free survival with a median time of 24 (range: 0.5-140.5) months. The causes of death were cytomegalovirus pneumonia (n=1), acute GVHD (n=1) and severe pulmonary infection (n=1).

CONCLUSIONIncreased-intensity of FBCA conditioning regimen could favor donor stem cell sustained engraftment for allo-HSCT in SAA.

Adolescent ; Adult ; Anemia, Aplastic ; therapy ; Child ; Child, Preschool ; Female ; Graft Survival ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Tissue Donors ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Vidarabine ; analogs & derivatives ; Young Adult