Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Background::The study aimed to describe the aortic valve morphology in Chinese patients underwent transcatheter aortic valve replacement (TAVR) for symptomatic severe aortic stenosis (AS), and the impact of sizing strategies and related procedural outcomes.Methods::Patients with severe AS who underwent TAVR were consecutively enrolled from 2012 to 2019. The anatomy and morphology of the aortic root were assessed. "Downsize" strategy was preformed when patients had complex morphology. The clinical outcomes of patients who performed downsize strategy were compared with those received annular sizing strategy. The primary outcome was device success rate, and secondary outcomes included Valve Academic Research Consortium-3 clinical outcomes variables based on 1-year follow-up.Results::A total of 293 patients were enrolled. Among them, 95 patients (32.4%) had bicuspid aortic valve. The calcium volume (Hounsfield Unit-850) of aortic root was 449.90 (243.15-782.15) mm 3. Calcium is distributed mostly on the leaflet level. Downsize strategy was performed in 204 patients (69.6%). Compared with the patients who performed annular sizing strategy, those received downsize strategy achieved a similar device success rate (82.0% [73] vs. 83.3% [170], P= 0.79). Aortic valve gradients (downsize strategy group vs. annular sizing group, 11.28 mmHg vs. 11.88 mmHg, P = 0.64) and percentages of patients with moderate or severe paravalvular regurgitation 2.0% (4/204) vs. 4.5% (4/89), P = 0.21) were similar in the two groups at 30 days after TAVR. These echocardiographic results were sustainable for one year. Conclusions::Chinese TAVR patients have more prevalent bicuspid morphology and large calcium volume of aortic root. Calcium is distributed mostly on the leaflet level. Compare with annular sizing strategy, downsize strategy provided a non-inferior device success rate and transcatheter heart valve hemodynamic performance in self-expanding TAVR procedure.

Hemophagocytic lymphohistiocytosis(HLH)is a life-threatening inflammatory response syndrome, which progresses rapidly.Its etiology and clinical manifestations are diverse, and its diagnosis is difficult, and the rescue treatment has not been unified at home and abrord.The paper reviews the classification, pathogenesis, clinical manifestations, laboratory indexs, diagnosis and treatment of HLH, in order to provide reference for timely diagnosis and individualized treatment.

Objective:To investigate the clinical value of muscle index changing value during neoadjuvant chemotherapy in predicting the prognosis of gastric cancer after radical gastrec-tomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 362 gastric cancer patients undergoing neoadjuvant chemotherapy combined with radical gastrectomy in 3 medical centers, including 163 cases in Fujian Medical University Union Hospital, 141 cases in the Affiliated Hospital of Qinghai University and 58 cases in St. Mary′s Hospital, from January 2010 to December 2017 were collected. There were 270 males and 92 females, aged from 26 to 79 years, with a median age of 61 years. Of 362 patients, 304 cases in Fujian Medical University Union Hospital and the Affiliated Hospital of Qinghai University were allocated into modeling group and 58 cases in St. Mary′s Hospital were allocated into validation group. Observation indicators: (1) changes of indicators including body composition parameters, tumor markers and stress status indicators in patients in modeling group during neoadjuvant chemotherapy; (2) follow-up and survival of patients; (3) analysis of risk factor affecting prognosis of patients in modeling group; (4) construc-tion and comparison of prognostic prediction models; (5) evaluation of prognostic prediction models. Follow-up was conducted using outpatient examination, telephone interview and mail communication to detect postoperative survival of patients up to April 2021. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Univariate and multivariate analysis were performed using the COX proportional hazard model. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. The Log-rank test was used for survival analysis. Results:(1) Changes of indicators including body composition parameters, tumor markers and stress status indicators in patients in modeling group during neoadjuvant chemotherapy: the subcutaneous adipose index, visceral adipose index, muscle index, carcinoem-bryonic antigen, CA19-9, body mass index, prognostic nutritional index and modified systemic inflammation score of 304 gastric cancer patients in the modeling group before neoadjuvant chemotherapy were 31.2 cm 2/m 2(range, 0.6?96.0 cm 2/m 2), 25.1 cm 2/m 2(range, 0.1?86.3 cm 2/m 2), 47.1 cm 2/m 2(range, 27.6?76.6 cm 2/m 2), 43.2 μg/L(range, 0.2?1 000.0 μg/L), 108.7(range, 0.6? 1 000.0)U/mL, 21.9 kg/m 2(range, 15.6?29.7 kg/m 2), 46.8(range, 28.6?69.0), 1.0±0.8, respectively. The above indicators of 304 gastric cancer patients in the modeling group before radical gastrec-tomy were 32.5 cm 2/m 2(range, 5.1?112.0 cm 2/m 2), 25.4 cm 2/m 2(range, 0.2?89.0 cm 2/m 2), 47.0 cm 2/m 2(range, 16.8?67.0 cm 2/m 2), 17.0 μg/L(range, 0.2?1 000.0 μg/L), 43.9 U/mL(range, 0.6?1 000.0 U/mL), 21.6 kg/m 2(range, 31.1?29.0 kg/m 2), 47.7(range, 30.0?84.0), 1.0±0.8, respectively. The changing value of above indicators of 304 gastric cancer patients in the modeling group during neoadjuvant chemotherapy were 1.4 cm 2/m 2(range, ?31.0?35.1 cm 2/m 2), 0.2 cm 2/m 2(range, ?23.5?32.6 cm 2/m 2), ?0.1 cm 2/m 2(range, ?18.2?15.9 cm 2/m 2), ?26.2 μg/L(range, ?933.5?89.9 μg/L), ?64.9 U/mL(range, ?992.1?178.6 U/mL), ?0.3 kg/m 2(range, ?9.7?7.1 kg/m 2), 0.9(range, ?27.1?38.2), 0.0±0.8, respec-tively. (2) Follow-up and survival of patients: 284 of 304 patients in the modeling group were followed up for 3 to 130 months, with a median follow-up time of 36 months. During follow-up, 130 cases died of tumor recurrence and metastasis and 9 cases died of non-tumor causes. The 5-year overall survival rate was 54.6%. Fifty-two of 58 patients in the validation group were followed up for 2 to 91 months, with a median follow-up time of 29 months. During follow-up, 21 cases died with the 5-year overall survival rate of 63.8%. (3) Analysis of risk factor affecting prognosis of patients in modeling group: results of univariate analysis showed that the postoperative pathological type and postoperative pathological staging were related factors affecting 5-year overall survival rate [ hazard ratio=1.685, 2.619, 95% confidence interval(CI): 1.139?2.493, 1.941?3.533, P<0.05] and 5-year progression free rate survival of 304 gastric cancer patients in the modeling group after radical gastrectomy ( hazard ratio=1.468, 2.577, 95% CI: 1.000?2.154, 1.919?3.461, P<0.05). Results of multivariate analysis showed that the postoperative pathological type and postoperative pathological staging were independent influencing factors for 5-year overall survival rate of 304 gastric cancer patients in the modeling group after radical gastrectomy ( hazard ratio=1.508, 2.287, 95% CI: 1.013?2.245, 1.691?3.093, P<0.05) and the postoperative patholo-gical staging was an independent influencing factor for 5-year progression free survival rate of 304 gastric cancer patients in the modeling group after radical gastrectomy ( hazard ratio= 2.317,95% CI: 1.719?3.123, P<0.05). (4) Construction and comparison of prognostic prediction models: the area under curve (AUC) of prognostic prediction model of subcutaneous adipose index changing value, visceral adipose index changing value, carcinoembryonic antigen changing value, CA19-9 changing value, body mass index changing value, prognostic nutritional index changing value, modified systemic inflammation score changing value for 304 gastric cancer patients in the modeling group were 0.549(95% CI: 0.504?0.593), 0.501(95% CI: 0.456?0.546), 0.566(95% CI: 0.521?0.610), 0.519(95% CI: 0.474?0.563), 0.588(95% CI: 0.545?0.632), 0.553(95% CI: 0.509?0.597), 0.539(95% CI: 0.495?0.584). The AUC of prognostic prediction model of muscle index changing value was 0.661(95% CI: 0.623?0.705) with significant differences to the AUC of prognostic predic-tion model of subcutaneous adipose index changing value, visceral adipose index changing value, carcinoembryonic antigen changing value, CA19-9 changing value, body mass index changing value, prognostic nutritional index changing value, modified systemic inflammation score changing value, respectively ( Z=3.960, 5.326, 3.353, 4.786, 2.455, 3.448, 3.987, P<0.05). The optimum cut-off value was 0.7 cm 2/m 2 for prognostic prediction model of muscle index changing. Kaplan-Meier survival curve showed there were significant differences of overall survival and progression free survival for gastric cancer patients with subcutaneous adipose index changing value <0.7 cm 2/m 2 and ≥0.7 cm 2/m 2 in the modeling group ( χ2 =27.510, 21.830, P<0.05). The nomogram prognostic prediction model was cons-tructed based on 3 prognostic indicators including muscle index change value combined with postoperative pathological type and postoperative pathological staging and the AUC of nomogram prognostic prediction model were 0.762(95% CI: 0.708?0.815) and 0.788(95% CI: 0.661?0.885) for the modeling group and the validation group, respectively. The AUC of postoperative pathological staging prognostic prediction model were 0.706(95% CI: 0.648?0.765) and 0.727(95% CI: 0.594?0.835)for the modeling group and the validation group, respectively. There were significant differences of the AUC between the nomogram prognostic prediction model of muscle index change value combined with postoperative pathological type and postoperative pathological staging and the postoperative pathological staging prognostic prediction model in the modeling group and the validation group, respectively ( Z=3.522, 1.830, P<0.05). (5) Evaluation of prognostic prediction models: the nomogram prognostic prediction model of muscle index change value combined with postoperative pathological type and postoperative pathological staging showed that patients with score of 0-6 were classified in the low risk group, patients with score of >6 and ≤10 were classified in the moderate-low risk group, patients with score of >10 and ≤13 were classified in the moderate-high risk group and patients with score of >13 were classified in the high risk group. Kaplan-Meier survival curve showed there were significant differences of the overall survival between the low risk group, moderate-low risk group, moderate-high risk group and high risk group patients in the modeling group and the validation group, respectively ( χ2 =75.276, 14.989, P<0.05). Results of decision making curve showed the nomogram prognostic prediction model of muscle index change value combined with postoperative pathological type and postoperative pathological staging had better clinical utility than the postoperative pathological staging prognostic prediction model in the modeling group and the validation group. Conclusions:The muscle index changing value of gastric cancer patient during neoadjuvant chemotherapy can be used as a prognostic indicator for gastric cancer patient prognosis after radical gastrectomy. The risk score of the nomogram prognostic prediction model of muscle index change value combined with postoperative pathological type and postoperative pathological staging can be used to evaluate the survival and prognosis of gastric cancer patients after radical gastrectomy.

Objective:To investigate the clinical efficacy of percutaneous endoscopic lumbar discectomy (PELD) with the aid of neurophysiological monitoring under general anesthesia.Methods:From August 2016 to October 2019, 58 patients underwent PELD under general anesthesia were selected in our hospital; 30 were via transformational approach and 28 were via interlaminar approach. The whole operative procedures were performed under continuous monitoring of spontaneous electromyography (SEMG), and the peak value, waveform and motor unit of SEMG at the surgical side were observed. The clinical outcomes were evaluated by visual analogue scale (VAS) and Oswestry disability index (ODI).Results:PELD was successfully performed in all 58 patients. Abnormal SEMG reactions were recorded in 8 patients (13.8%), manifested as clustered or frequent high amplitude action potentials; 5 patients (16.7%, 5/30) were via transformational approach and 3 (10.7%, 3/28) were via interlaminar approach. Two patients relapsed at 3 and 6 weeks after surgery, respectively; one was treated with PELD again and the other one was treated with lumbar fusion and instrument fixation. The pain at the lumbago and leg was alleviated obviously after surgery in the 56 patients; the VAS scores were 7.43±1.32, 2.55±0.87 and 1.59±0.87 before surgery, and 3 d and 3 months after surgery, respectively, with significant differences ( P<0.05); the mean ODI before surgery and 3 months after surgery were 67.36±7.13 and 12.39±5.48, respectively, with significant difference ( P <0.05). Conclusion:PELD with the aid of neurophysiological monitoring under general anesthesia is safe and reliable, which can achieve good clinical efficacy.

Objective: To observe the effect of cinobufacin on the expression of phosphorylated PTEN protein in human breast cancer MCF-7 and MDA-MB-231 cells. Methods: The MCF-7 and MDA-MB-231 cells were divided into control group, high dose group and low dose group according to random number table method. Low and high dose groups were added with 0.5, 5.0 mg/ml cinobufacin, 100 μl/well for intervention, respectively, while the control group was added with equal volume of RPMI-1640 medium. Cell Counting Kit-8 was used to detect cell proliferation assay at 24, 48, and 72 h following intervention. Immunoblotting was used to detect the expression of AKT/mTOR pathway protein expression and p-PTEN. Results: At 24, 48, and 72 h after intervention, the proliferation of MCF-7 and MDA-MB-231 cells in high dose group were significantly decreased (P<0.05). Compared with the control group, the expression of p-AKT (MCF-7: 0.357 ± 0.064, 0.215 ± 0.056 vs. 0.924 ± 0.085; MDA-MB-231: 0.310 ± 0.022, 0.194 ± 0.019 vs. 0.811 ± 0.089), p-mTOR (MCF-7: 0.476 ± 0.039, 0.217 ± 0.038 vs. 0.838 ± 0.058; MDA-MB-231: 0.300 ± 0.031, 0.223 ± 0.025 vs. 0.896 ± 0.096), p-S6 (MCF-7: 0.551 ± 0.068, 0.428 ± 0.041 vs. 1.254 ± 0.264; MDA-MB-231: 0.281 ± 0.014, 0.197 ± 0.012 vs. 0.748 ± 0.022), p-PTEN (MCF-7: 0.487 ± 0.170, 0.184 ± 0.135 vs. 1.003 ± 0.284, P<0.05; MDA-MB-231: 0.261 ± 0.184, 0.170 ± 0.105 vs. 1.014 ± 0.206) in the low and high dose group significantly decreased (P<0.05). Conclusions Cinobufacin inhibited the proliferation of MCF-7 and MDA-MB-231 cells, which may be mediated by down-regulation of the expression of the upstream of the AKT/mTOR signaling pathway p-PTEN.

Objective To investigate the effect of salidroside on xenograft tumor of human pancreatic cancer cells BxPC-3 in nude mice.Methods BxPC-3 subcutaneous xenograft tumor nude mice model was established,and the mice were randomly divided into control group,salidroside 25 mg group and salidroside 50 mg group using random number method (6 mice in each group).25 mg/kg and 50 mg/kg salidroside by intragastric administration was performed per day for 3 weeks and the tumor was resected.The tumor was weighed and the diameter was measured.Cell apoptosis was detected by Tunel staining.Results As result,tumor weight was (1.561 ± 0.416)g in the control group,(0.742 ± 0.272)g in the 25 mg group and (0.276 ± 0.064) g in the 50 mg group respectively;tumor volume was (2354.35 ± 523.11) mm3,(991.43 ± 348.91) mm3 and (403.60 ± 130.98) mm3,respectively;and the apoptosis rate of the transplanted tumor cells was (23.74 ± 5.88) ％,(49.30 ± 6.75) ％ and (64.97 ± 6.99) ％,respectively,with significant difference among three groups (all P ＜ 0.01).The effect of salidroside on inhibiting proliferation and promoting apoptosis was dose dependent.Conclusions Salidroside can significantly inhibit the proliferation and growth of pancreatic cancer BxPC-3 cells via promoting cell apoptosis and inhibiting cell proliferation.

Objective To evaluate the efficacy and safety of bronchoalveolar lavage( BAL) in the treatment of neonatal atelectasis with fiberoptic bronchoscopy under ultrasound monitoring. Methods From June 2018 to December 2018,29 children were diagnosed as atelectasis by lung ultrasound. After conventional mechanical vibration and sputum ineffective,BAL was treated with fiberoptic bronchoscopy. All patients be-fore operation were monitored by ultrasound to find the lung segment where the atelectasis was located. 0. 9%NaCl solution was injected by fibrobronchoscope(1～2 ml/kg),and then sucked to ensure the recovery rate of the lavage fluid was more than 50%. After each lavage,ultrasound was immediately used to monitor the recovery of atelectasis to determine whether or not to continue the lavage. One course of treatment could be continuously performed BAL 1 to 3 times a day,1 course per day,and up to 3 courses of lavage. We analyzed the efficacy,adverse reactions and complications of BAL in the treatment of neonatal atelectasis under ultra-sound monitoring. Results Twenty-nine patients underwent BAL treatment with fiberoptic bronchoscopy,25 cases (86. 2%) were cured,3 cases (10. 3%) were effective,and 1 case (3. 4%) was ineffective. All chil-dren had stable vital signs during treatment. Among them,11 cases (37. 9%) had transient hypoxemia, 3 ca-ses (10. 3%) had tracheal mucosal injury, and 2 cases (6. 9%) had hoarseness. There were no serious com-plications such as pulmonary hemorrhage,pneumothorax,and cardiac arrest. Conclusion BAL treatment of atelectasis under lung ultrasound monitoring has obvious effect,easy to operate,no radiation,no obvious ad-verse reactions and complications,which is worthy of clinical application.

Inhibition of NEDD8 activating enzyme (NAE),the rate limiting enzyme in Neddylation,can suppress the activity of ubiquitin-proteasome system (UPS) pathway and decrease degradations of related proteins,resulting in cell apoptosis.Compared with the proteasome inhibitors,NAE inhibitors can interfere with cellular homeostasis with more specificity.With the rapid development of the research on NAE,a variety of NAE inhibitors have been reported.According to the structure characteristics,NAE inhibitors can be divided into the AMP analogues,double flavonoids deoxidization duckbill alkali ketone derivatives and metal rhodium complex classes,etc.The process of ubiquitinated modified Neddylation,NAE and physiological role of tumor has been introduced in this paper,and the latest progress of NAE inhibitors also has been summarized.

AIM: To investigate the effect of cobalt chloride (CoCl2) on the apoptosis of neural stem cells (NSCs) and the expression of microRNA-26a (miR-26a) in vitro, and to explore the mechanisms of NSC apoptosis in-duced by CoCl 2 .METHODS:NSCs were exposed to CoCl 2 at different doses (200~600μmol/L) for 24 h.The cell via-bility and apoptosis were measured by CCK-8 assay and TUNEL method.The expression of miR-26a-3p, miR-26a-5p, GSK-3β, caspase-3, Bcl-2 and Bax was examined by real-time PCR.The protein levels of Bcl-2 and Bax were detected by Western blotting .RESULTS: The cell viability was inhibited and the apoptosis of NSCs was increased significantly by CoCl2 in a dose-dependent manner (P<0.05).CoCl2 at concentration of 400μmol/L for 24 h was used to induce apopto-sis and the expression of miR-26a was down-regulated compared with control (P<0.05).Exposure to CoCl2 at concentra-tion of 400μmol/L up-regulated the expression of GSK-3β, caspase-3 and Bax , down-regulated the expression of Bcl-2 and Bcl-2/Bax (P<0.05).CONCLUSION:CoCl2 at concentration of 400μmol/L induces the apoptosis of NSCs obviously . CoCl2 may induce the NSC apoptosis by mitochondrial apoptotic pathway .Declining miR-26a may be related to NSC apopto-sis.