Objective To investigate the therapeutic effect of NSR siRNA on nerve regeneration following spinal cord hemi-transsection injury in rats. Methods Rats with T8 spinal cord hemi-trans-section were didded into 3 groups, ie, siRNA group, NS group and control group. SiRNA or NS was in-jected into lateral cerebral ventricle just after spinal cord injury. The therapeutic effect of NgR siRNA was evaluated by using BBB locomotor rating scale, retrograde horseradish peroxidase(HRP)tracing and HE staining. Results BBB locomotor rating scale showed that the recovery of the locomotor function of siRNA group seemed to be better than that of the other two groups from the 4th week, but there was no statistical difference. Retrograde HRP tracing showed a large number of positive cells in the anterior horn of spinal cord, with statistical difference compared with NS group and control group(P<0. 05). Eight weeks after spinal injury, HE staining showed disorderly distribution of the fibres in NS group and control group but serial fibres in the injury region in siRNA group. Conclusion NSR siRNA may promote the nerve regeneration following spinal cord injury.

AIM: To determine the low content of Ophiopogon japonicus saponin metabolite in vivo. METHODS: HPLC-MS method of determining rat's metabolite diosgenin in vivo was established after single-dose oral Ophiopogon japonicus saponin enteric microsphere. RESULTS: The detection limit was about 50 ng/mL,Linearity,precision of intra and inter-day and reproducibility of the method were good. CONCLUSION: This method accords with the analysis requirement.It can give an effective measure and foundation for studying TCM saponin bioavailability.

Objective To compare the difference in cardiac injuries between asphyxia and ventricular fibrillation modes in different periods after cardiac arrest (CA).Methods The model was established in Cardiopulmonary Resuscitation Lab,Sun Yat-sen University.A total of 35 male SD rats were used to produce the asphyxia or ventricular fibrillation (VF) cardiac arrest models randomly.Both of the two modes were induced 8 minutes cardiac arrest.The myocardial HE stains,mitochondrial respiratory control ratio (RCR),and echocardiography were observed at 4 h,24 h and 72 h after ROSC (restoration of spontaneous circulation).The results were expressed as (-x ± s),t test was performed to compare between two groups,and one way analysis of variance was used to compare multiple groups.P ＜ 0.05 was considered as significant difference.Results HE stains showed damages were more serious in the VF mode than in asphyxia mode at 4 h,and both of them had a disorderly-arranged myocardium at 72 h.RCR in VF mode became worse at 4 h,and RCR resumed at 24 h in both modes without significant difference compared with the sham operated rats.The echocardiography showed VF mode had a lower left ventricular ejection fraction (LVEF) than asphyxia mode at 4 h (29.68％ vs.42.16％,P =0.03),and there was no difference in LVEF between VF mode and the sham operated rats at 24 h,however no difference in LVEF between the asphyxia and sham operated rats at 72 h.Both of them had a thicker left ventricular anterior wall than the sham operated rats at 72 h (2.41 mm vs.1.72 mm,P=0.013; 2.61 mmvs.1.72 mm,P=0.007),and there was no significant difference between them.Conclusions The ventricular fibrillation mode has a more severe injuries in early period,but it recovers sooner than asphyxia one.Both of two groups get compensatory left ventricular hypertrophy in later period of ROSC.

Objective To investigate the protective role of carbon monoxide releasing molecule-2 (CORM-2) in post-resuscitation myocardial dysfunction (PRMD) in rat models of cardiopulmonary resuscitation (CPR).Methods Cardiopulmonary resuscitation model was established after cardiac arrest induced by ventricular fibrillation.Male healthy Sprague-Dawley (SD) rats were randomly (random number) divided into 4 groups according to random number table:control group,CORM-2 group,inactive CORM-2 (iCORM-2) group and Sham group,in which the equal volume (1 mL) of 0.2％ DMSO,50 μmol/kg CORM-2,50 μmol/kg iCORM-2 and 0.2％ DMSO were respectively administered into the rats of these groups after resuscitation.The ejection fraction (EF) of left ventricle and myocardial performance index (MPI) were measured to detect the myocardial function by echocardiography at 12 hours after resuscitation.Mitochondrial respiration was assessed with Clark oxygen electrode at the same time.Western blot was used to determine the ratio of mitochondrial cytochrome c (cyt c) to cytoplasmic cyt c as well as caspase-3 level.Multiple comparisons were made by analysis of variance.Results Compared with the control group,higher EF and MPI,higher state Ⅲ respiration rate and respiratory control rate (RCR) of mitochondria,and decreased ratio of mitochondrial cytc/cytoplasmic cyt c and lower caspase-3 level were observed in the CORM-2 group (P ＜ 0.05).However,there were no significant differences in above biomarkers found between iCORM-2 group and control group (P ＞ 0.05).Conclusions The CO released from CORM-2 might improve mitochondrial respiration and PRMD by inhibition of myocardial apoptosis via a mitochondrial pathway.

Objective To observe the dynamic changes of myocardial structure and dysfunction during post-resuscitation period in order to establish a rat mode of post-resuscitation myocardial dysfunction after cardiac arrest resulted from electric stimulation-induced ventricular fibrillation (VF) and cardiopulmonary resuscitation (CPR).Methods A total of 40 male Sprague-Dawley (SD) rats were randomly (random number) assigned into post-resuscitation (PR) 4 h,PR 12 h,PR 24 h,PR 72 h and sham groups.VF was induced by an alternating electric current delivered to the right ventricular endocardium and untreated for 8 min.Biphasic waveform defibrillation was attempted and mechanical ventilation was synchronized after 6 min of CPR.Myocardial function was assessed with serum myocardial enzyme activity,echocardiography,mitochondrial respiratory function and histopathologic findings at different intervals.Results Thirty-two animals were successfully resuscitated with restoration of spontaneous circulation (ROSC) in 86％ (32/37) rats.Compared with sham group,severe systolic and diastolic heart failure were found at 4 h after ROSC and then gradually improved without significant difference (P ＞0.05) in ejection fraction at PR 72 h after ROSC was found,whereas thickened ventricular wall and increased myocardial performance index as well as interstitial proliferation were observed at 72 h after ROSC.Conclusions A rat model of post-resuscitation myocardial dysfunction after cardiac arrest resulted from electric stimulation-induced VF and CPR was successfully established.

Objective To study the relative bioavailability after ig rats with Ophiopogon japonicus saponin enteric microsphere in single-dose so as to give directly preparation quality evaluation. Methods(Establishing the) analysis method of HPLC-MS and dealing with the blood sample by solid phase extraction. Determining the diosgenin concentration in plasma after ig Ophiopogon japonicas saponin enteric microsphere and the suspension as comparison in single-dose, and calculating pharmacokinetic parameters and relative bioavailability. Results C_(max), t_(max), and AUC_(0-72) were (637.81?17.23) ng/mL, (4.0?0.0) h, and 7 840.01?412.03, respectively. Conclusion The characteristics of absorption, distribution, and elimination are resembled in rats between two preparations by ig in single-dose. The pharmacokinetic parameters are also identical. The relative bioavailability is (91.10?3.44)%.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Objective:To evaluate the effect of the timing of peripancreatic drainage on the survival outcome of patients with severe acute pancreatitis (SAP).Methods:This retrospective study included 271 patients with SAP admitted to two tertiary hospitals from January 2015 to December 2019. The Acute Physiology and Chronic Health EvaluationⅡ score (APACHEⅡ), Sequential Organ Failure Assessment score (SOFA), computed tomography (CT) grade, peripancreatic drainage situations, and survival outcome of patients were recorded. Patients were divided into the early and non-early peripancreatic catheter drainage groups (EPCD and non-EPCD). The data were analyzed using the Cox proportional hazard model for propensity score matching (PSM) and stratification.Results:After PSM, the 30-day and 90-day risk of death between the EPCD and non-EPCD groups were significantly different (0.134, 95% CI: 0.029-0.576, P=0.007; 0.166, 95% CI: 0.044-0.631, P=0.008, respectively). Furthermore, stratified analysis revealed significant differences in 30-day and 90-day risk of death between the EPCD and non-EPCD groups when the SOFA score was≥4 or the APACHEⅡ score was ≥8. Conclusions:For patients with SAP with SOFA score ≥4 or APACHEⅡ score≥8, early peripancreatic drainage can reduce the risk of death, but CT grading is not helpful for the decision-making of drainage timing in patients with SAP.

Objective:To analyze the coronary angiographic (CAG) characteristics of coronary artery lesion (CAL) in children with Kawasaki disease (KD), and to clarify the necessity of CAG in the diagnosis and treatment of KD combined with CAL in children.Methods:It was a retrospective study to analyze the clinical data, electrocardiogram, echocardiography, time and findings of CAG in children with KD and CAL who underwent CAG in Shanghai Children′s Medical Center of Shanghai Jiao Tong University School of Medicine from January 2013 to August 2022.The distribution, type, severity, and prognosis of CAL were analyzed.Results:A total of 117 children with KD and CAL were included in the analysis.The onset age of KD was from 2 months to 12.8 years old, and the age of performing CAG was from 8 months to 18.1 years old.A total of 234 coronary artery lesions were detected in 117 cases.Among them, CAL in the right coronary artery (RCA), left anterior descending branch (LAD), left main coronary artery and left circumflex artery were detected in 96 branches(41.1%), 78 branches(33.3%), 44 branches(18.8%), and 16 branches(6.8%), respectively.Unilateral coronary artery involvement was detected in 43 cases (36.8%), of which LAD was the dominant; while bilateral involvement was detected in 74 cases (63.2%), among which, LAD and RCA were the most involved arteries.Stratified by the degree of coronary involvement, large coronary aneurysms and severe coronary stenosis were most frequently occurred in the RCA and LAD.In contrast, 10 cases (13.6%), 20 cases (24.3%), 55 cases (45.8%) and 37 cases (67.3%) of intraluminal lesions were found in small, medium and large coronary aneurysms, and stenosis or occlusion, respectively.The incidence of intraluminal lesions tended to be higher in the site of severe lesions.CAG showed stenosis or occlusion in a total of 55 cases, and collateral circulation at varying degrees was found in cases of severe stenosis or occlusion.Conclusions:CAL in children with KD are complex and varied.Although clinical symptoms, routine electrocardiogram and cardiac ultrasound may indicate severe CAL.Their applications are limited by the diagnosis of the type (especially stenosis), degree, and extent of CAL, as well as the detection of extracoronary lesions.CAG is of great significance to identify vascular lesions and guide clinical management of KD combined with CAL in children.

Objective·To analyze the progression of children with severe coronary artery lesions due to Kawasaki disease by coronary artery angiography,and evaluate the diagnostic value of echocardiography in these children.Methods·A retrospective analysis was performed to enroll children with Kawasaki disease whose coronary artery lesions were graded Ⅳ or above from Shanghai Children's Medical Center,Shanghai Jiao Tong University School of Medicine,from January 2013 to January 2023.The subjects were required to have received at least 2 times of coronary angiogram,and their clinical and imaging data were collected to analyze the progression of the lesions.Echocardiography results were compared with the results of the coronary angiogram.Results·A total of 21 children were included,including 15 males and 6 females,with a median age at onset of 3 years and 6 months,a median age at initial coronary angiography of 7 years and 11 months,a median interval of 4 years and 5 months between the time of onset and initial angiography,a median age at angiographic review of 9 years and 2 months,and a median interval of 1 year and 3 months between the time of initial angiography and review.Coronary stenosis or occlusion was detected in 13 children in the initial angiography,of whom 6 underwent coronary artery bypass grafting(CABG)and had their angiography reviews 1 year later.The review results showed that the bridging vessels were unobstructed and no obvious stenosis was observed.Fifteen children had progression of the lesions detected by echocardiography in the subsequent follow-up and had their angiogram reviews,of whom 8 had significant progression of the coronary lesions.Intracoronary balloon dilatation was performed in 1 case,and CABG was performed in another case.Sixteen lesions of coronary stenosis or occlusion were detected in the initial angiography in 21 children,while only 1 lesion of coronary stenosis was detected by echocardiography during the same period of time.Twenty-eight medium-to large-sized coronary aneurysms were detected in the initial angiography in the 21 children,and the diameters of the 28 aneurysms measured by echocardiography and coronary angiogram were subjected to the Bland-Altman analysis.The Bland-Altman analysis showed that the difference in maximum diameter between 2 methods was(1.63±2.33)mm,with 95％CI of-2.95-6.21 mm.Conclusion·Coronary artery lesions due to Kawasaki disease may be progressive;in the children with severe lesions,coronary artery stenosis or occlusion may be missed or misdiagnosed and some errors may exist in the measurement of diameters of aneurysms by echocardiography.Regular review of coronary angiography is needed.