In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; administration & dosage ; genetics ; immunology ; Capsid Proteins ; administration & dosage ; genetics ; immunology ; Female ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Herpesvirus 1, Suid ; genetics ; metabolism ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Swine ; Swine Diseases ; immunology ; prevention & control ; virology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Objective:To explore the risk factors of hypopituitorism after aneurysmal subarachnoid haemorrhage (aSAH) in the acute phase and analysis the effect of hypopituitorism on prognosis.Methods:Patients with aSAH that were diagnosed and treated in China-Japan Union Hospital from Sep. 2017 to Sep. 2018 and undergoing pituitary function evaluation within 3 to 7 days were retrospectively analyzed. 72 patients were enrolled, including 31 males and 41 females. The average age was 50.1 years old (36-71) . The WFNS gradings were Ⅰ, 24; Ⅱ, 20; Ⅲ, 15; Ⅳ, 10; Ⅴ, 3. The Fisher gradings were 2, 21; 3, 38; 4, 13. Acute hydrocephalus happened in 13 cases. Aneurysm located in the Willis circle in 56 cases and not in the Willis circle in 16 cases. 37 cases were treated by microsurgical clipped and 35 cases were treated by embolism. Patients with hypocortisolism were treated by hydrocortisone replacement, and patients with hypothyroidism were given levothyroxine replacement therapy. Patients were followed up at 3 months and their recovery was evaluated by GOS score.Results:Hypopituitorism was detected in 34 patients; the incidence of hypopituitorism was 47.2%. There was no significant correlation between the occurrence of hypopituitorism and age, gender, and hydrocephalus ( P>0.05) . Patients with WFNS grade ≥Ⅳ, Fisher grade 4, aneurysm located in the Willis circle, and treated by clipping were more likely to undergo hypopituitorism. The proportion of patients with good recovery (GOS ≥4) in the hypopituitorism group was smaller than that in the normal pituitary function group at the 3-month follow-up. Conclusions:Patients with WFNS grade ≥Ⅳ, Fisher grade 4, aneurysm located in the Willis circle, and treated by clipping are more likely to undergo hypopituitorism. Hypopituitorism in acute stage affects the prognosis of aSAH patients. Patients with hypocortisolism and hypothyroidism should be treated actively.

With the deepening of the research on the "two sides" of arsenic from poison to medicine, arsenic has attracted extensive attention in affecting programmed cell death (PCD) and causing damage to a variety of organs. Recent studies have showed that reactive oxygen species (ROS) produced by intracellular arsenic metabolism is closely related to PCD induction. However, the specific mechanism is still unclear. In this paper, we have reviewed the main PCD forms, such as apoptosis, autophagy and necroptosis induced by arsenic via ROS and their possible mechanisms, in order to provide basic information for further research and prevention of arsenic toxicity, which is helpful for clinical development and utilization of arsenic in the treatment of tumors and related diseases.

Objective:To observe the role of nuclear factor erythroid 2-related factor 2 (Nrf2) in regulating apoptosis during malignant transformation of human bronchial epithelial cells (HBE cells) induced by sodium arsenite (NaAsO 2). Methods:HBE cells were treated with 0.0 and 1.0 μmol/L NaAsO 2, which were control group and arsenic exposed group respectively. HBE cells were treated with 1.0 μmol/L NaAsO 2 for 43 passages to establish a malignant transformation model. The dynamic changes of indexes in different passages (0, 1st, 8th, 15th, 22nd, 29th, 36th, and 43rd) after exposure to NaAsO 2 were monitored, including the apoptosis rate detected by flow cytometry and apoptosis-related proteins and Nrf2 protein detected by Western blotting. Nrf2 siRNA was transfected into malignant transformed HBE cells (T-HBE cells) to silence Nrf2. The silencing effect of Nrf2 protein was verified. And, the apoptosis rate and apoptosis-related proteins were detected. Results:With the increase of arsenic exposure, the apoptosis rates of HBE cells decreased (0, 1, 8, 15, 22, 29, 36 and 43 passages were 0.370 ± 0.029, 0.443 ± 0.069, 0.357 ± 0.046, 0.330 ± 0.016, 0.273 ± 0.050, 0.160 ± 0.024, 0.110 ± 0.022, 0.097 ± 0.012, respectively, Ftrend = 22.981, P < 0.05). Compared with the 0 passage cells, the apoptosis rates of the 22nd, 29th, 36th and 43rd passages in the arsenite group were lower. The differences between them were statistically significant ( P < 0.05). With the increase of arsenic exposure, the expressions of pro-apoptotic proteins caspase-3, cleaved-caspase-3, C/EBP-homologous protein (CHOP) and B-cell lymphoma-2 (Bcl-2) associated X protein (Bax) showed downward trends ( Ftrend = 22.356, 3.738, 6.130, 8.061, P < 0.05), while the anti-apoptotic proteins myeloid cell leukemia 1 protein (Mcl-1) and Bcl-2 showed upward trends ( Ftrend = 58.201, 7.691, P < 0.05). Compared with the 0 passage and the control group of the same passage, from the 22nd passage of caspase-3, cleaved-caspase-3, from the 15th passage of CHOP, Mcl-1, and Bcl-2, from the 29th passage of Bax in the arsenite group, the differences of protein were statistically significant ( P < 0.05). However, there were no significant differences in caspase-8, cleaved-caspase-8, caspase-12 and cleaved-caspase-12 protein expressions in the arsenic group ( P > 0.05). Compared with the 0 passage and the control group of the same passage, from the 8th passage of Nrf2 proteins in the arsenite group, the differences of expressions were statistically significant ( P < 0.05). Compared with T-HBE cells transfected with Con siRNA (control), the apoptosis rate of T-HBE cells transfected with Nrf2 siRNA was higher ( P < 0.05). Compared with T-HBE cells transfected with Con siRNA, the expression levels of Nrf2, Bcl-2 and Mcl-1 in T-HBE cells transfected with Nrf2 siRNA were lower ( P < 0.05), while the expression levels of cleaved-caspase-3/caspase-3, caspase-3, cleaved-caspase-3, CHOP, and Bax were higher ( P < 0.05). Conclusion:Nrf2 may regulate mitochondrial apoptotic pathway through Bcl-2, Mcl-1 and Bax, and endoplasmic reticulum apoptotic pathway through CHOP, so as to inhibit the apoptosis of HBE cells and participate in the process of malignant transformation of HBE cells induced by NaAsO 2.

Objective:To explore the role of nuclear transcription factor erythrocyte line-2p45 (NF-E2) related factor-2 (NRF2) on autophagy during malignant transformation of immortalized human keratinocytes (HaCaT) induced by sodium arsenite (NaAsO 2). Methods:Using cell culture methods, long-term cultured HaCaT cells in DMEM high-glucose medium containing 0.0 (control group) and 1.0 μmol/L NaAsO 2 (arsenic-exposed group) to the 35th generation were used to construct a cell malignant transformation model, and 0, 1, 7, 14, 21, 28 and 35th generation cells of control group and arsenic-exposed group were collected during establishment of cell malignant transformation model. The NRF2 siRNA, phosphatidylinositol-3-hydroxykinase (PI3K) inhibitor LY294002 and mammalian target of rapamycin (mTOR) inhibitor Rapamycin were used to treat the 35th generation of malignant transformed HaCaT cells in arsenic-exposed group (T-HaCaT). The protein expressions of NRF2, PI3K-protein kinase B (Akt)-mTOR signaling pathway related indicators PI3K, Akt, mTOR, phosphorylated (p)-PI3K, p-Akt, p-mTOR, autophagy-related proteins p62, Beclin1, microtubule-associated protein-1 light chain (LC)3Ⅰ, and LC3Ⅱof different generations HaCaT cells in control group and arsenic-exposed group, and T-HaCaT cells of each treatment group were determined by Western blotting. Results:There were significant differences in the NRF2 protein and the ratios of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR between different generations HaCaT cells in arsenic-exposed group ( F = 9.371, 16.035, 15.932, 27.739, P < 0.05), and they were higher than NRF2 protein and ratio of p-mTOR/ mTOR of the same generation in control group ( P < 0.05). Compared with HaCaT cells of the same generation, the expressions of NRF2, p-PI3K, p-Akt, p-mTOR and p62 proteins in T-HaCaT cells were significantly higher, Beclin1 protein expression and the ratio of LC3Ⅱ/LC3Ⅰ were significantly lower ( P < 0.05). The NRF2 silenced T-HaCaT cells had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expressions of NRF2, p-mTOR and p62 than the corresponding control siRNA (Con siRNA) group ( P < 0.05). The T-HaCaT cells in LY294002 treatment group had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expressions of NRF2, p-PI3K, p-Akt and p-mTOR proteins than the corresponding non-treatment group ( P < 0.05). The T-HaCaT cells in Rapamycin treatment group had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expression of p-mTOR protein than the corresponding non-treatment group ( P < 0.05). Conclusions:During the arsenic-induced malignant transformation of HaCaT cells, NRF2 can act as a downstream factor of PI3K-Akt and an upstream factor of mTOR in PI3K-Akt-mTOR signaling pathway, an important regulatory mechanism of autophagy. This abnormal expression of autophagy may eventually lead to malignant transformation of cells.

Objective:To evaluate the correlation between endemic arsenic poisoning and abnormal electrocardiogram (ECG).Methods:PubMed, Web of Science, Embase, China National Knowledge Infrastructure (CNKI), Wanfang data, VIP and other databases were used for literature retrieval, and epidemiological literatures related to abnormal ECG of endemic arsenic poisoning published in domestic and abroad were included in the study. The time limit was from the establishment of the database to December 1, 2020. RevMan 5.3 was used for Meta-analysis of binary variables. Random effect model was selected according to the results of heterogeneity, and odds ratio ( OR) was used as the effect index. Characteristic changes were found by subgroup analysis. Bias was published by funnel plot. Results:Nine articles were included in this Meta-analysis, with 6 articles in Chinese and 3 articles in English, respectively. The abnormal ECG changes included QTc prolongation, ST-T segment change, left axis deviation and arrhythmia. Finally, 1 975 cases were included in the exposure group, including 575 cases of abnormal ECG; 750 cases of control group, including 145 cases of abnormal ECG. Meta-analysis showed that the combined OR value [95% confidence interval ( CI)] of abnormal ECG changes was 4.41 (2.83 - 6.87), with statistical significance between the two groups ( Z = 6.56, P < 0.05); the results of subgroup analysis showed that the combined OR values (95% CI) of QTc prolongation, ST-T segment change, left axis deviation and arrhythmia were 12.30 (5.91 - 25.59), 2.74 (1.39 - 5.41), 2.93 (0.89 - 9.62) and 4.13 (2.38 - 7.17), respectively. Conclusions:Endemic arsenic poisoning may cause abnormal ECG. Prolongation of QTc caused by arsenic exposure may be the characteristic change of abnormal ECG.

Objective:To systematically evaluate the correlation between maternal arsenic exposure during pregnancy and congenital heart disease (CHD) in offspring.Methods:Literature search was performed through databases such as PubMed, Web of Science, Embase, China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and VIP.com to include epidemiological literature on association between maternal arsenic exposure during pregnancy and offspring CHD published domestically and internationally. The search was conducted from database establishment until November 2, 2022. Stata MP15 software was used for meta-analysis of binary variables, and I 2 statistics and Q test were used for heterogeneity test, fixed effect model or random effect model was selected based on the test results. Using OR value (95% CI) as the effect evaluation indicator, subgroup analysis was conducted based on CHD subtypes [conotruncal defects (CTD), atrioventricular septal defect (AVSD), total anomalous pulmonary venous return (TAPVR), left ventricular outflow tract obstruction (LVOTO), right ventricular outflow tract obstruction (RVOTO), atrial septal defect (ASD)/ventricular septal defect (VSD), and patent ductus arteriosus (PDA)]. Results:Nine articles were finally included, including two Chinese and seven English articles. Among them, 8 articles had CHD as the outcome, 5 articles had ASD/VSD as the outcome, 4 articles had CTD as the outcome, 3 articles had LVOTO as the outcome, 2 articles had PDA as the outcome, and 1 article had RVOTO as the outcome. An analysis was conducted on 8 articles with CHD as the outcome. After heterogeneity testing, I 2 = 88.5% and P < 0.001, indicating significant heterogeneity. A random effect model was used for meta-analysis, and the combined OR value (95% CI) was 1.51 (1.40 - 1.62). The results of CHD subgroup analysis showed that the combined OR values (95% CI) for ASD/VSD, CTD, LVOTO, PDA, and RVOTO were 1.68 (1.53 - 1.84), 1.64 (1.29 - 2.09), 2.89 (1.82 - 4.61), 1.78 (1.53 - 2.08), and 0.81 (0.64 - 1.03), respectively. Conclusion:Maternal arsenic exposure during pregnancy is associated with development of offspring CHD, including ASD/VSD, CTD, LVOTO, and PDA as the common lesions in offspring CHD.

Objective To evaluate the clinical effect of minimally invasive aortic valve replacement with Perceval sutureless aortic bioprosthesis in upper ministernotomy or right anterior thoracotomy. Methods From March to November 2022, the patients with simple aortic valve disease were enrolled in the Department of Cardiovascular Surgery of West China Hospital, Sichuan University. After preoperative evaluation, Perceval sutureless bioprosthesis was successfully used to perform aortic valve replacement through the upper ministernotomy or right anterior thoracotomy. The perioperative clinical data and ultrasonic measurement data of all patients were recorded. Results A total of 5 patients with simple aortic valve disease were included, including 3 females and 2 males, with a mean age of 71.2 years. Perceval sutureless bioprosthesis was successfully implanted in 5 patients, with a success rate of 100%. There were 3 patients receiving upper ministernotomy and 2 patients receiving right anterior thoracotomy. Two patients underwent ascending aortic plasty at the same time. The mean cardiopulmonary bypass time was 61.0 min, and aortic cross-clamping time was 32.2 min. All patients were discharged successfully without perivalvular leakage, atrioventricular block or stroke. Conclusion The implantation method of Perceval sutureless bioprosthesis is simple, which can effectively reduce the perioperative risk by shortening the overall operation time, cardiopulmonary bypass time and aortic cross-clamping time. At the same time, its clinical application has promoted the development and popularization of minimally invasive aortic valve replacement, which together with Perceval sutureless bioprosthesis effectively combinates surgical effect and minimally invasive treatment, and has a good clinical application prospect because of its reliable safety and effectiveness.

Objective To evaluate the clinical efficacy of two-stage retrograde hybrid repair for acute aortic dissection involving the aortic arch complicated with distal malperfusion syndrome. Methods From May 2019 to December 2022, the patients presented with acute aortic dissection involving the aortic arch complicated with distal malperfusion syndrome treated in the Department of Cardiovascular Surgery of West China Hospital, Sichuan University were enrolled. After preoperative evaluation, all patients underwent priority emergency interventional surgery to improve distal malperfusion, and then underwent two-stage hybrid surgery to repair proximal aortic lesions. The perioperative clinical and imaging data were retrospectively analyzed. Results Five patients were collected, including 4 males and 1 female, with a median age of 58 years. The main manifestations were lower limb ischemia and renal insufficiency in 3 patients, and poor intestinal perfusion in 2 patients. All patients were given priority to interventional surgery to implant graft stents or bare stents and necessary branch artery intervention, and then successfully performed two-stage hybrid surgery, including type Ⅰhybrid surgery for 2 patients, type Ⅱ hybrid surgery for 1 patient and type Ⅲ hybrid surgery for the other 2 patients, with a success rate of 100.0%. All patients were discharged successfully, and the function of the organs with poor perfusion returned to normal. Only 1 patient recovered to grade 4 muscle strength of the diseased lower limbs upon discharge. No adverse events such as amputation, exploratory laparotomy and intestinal resection or long-term hemodialysis occurred. Conclusion The application of two-stage retrograde hybrid repair in the surgical treatment of acute aortic dissection involving the aortic arch complicated with distal malperfusion syndrome is safe and effective, and is helpful to improve the perioperative survival rate, and clinical outcomes of such patients.