Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.

Objective To analyze the feasibility of local LINGO-1 polyclonal antibody administration for treatment of spinal cord injury in adult rats. Methods Twenty-four adult female SD rats were randomized into sham-operated group, rabbit IgG group and LINGO-1 antibody group. In the latter two groups, partial transaction of the T9 segment of the spinal cord was performed to completely sever the dorsal eorticospinal tract, followed immediately by administration of rabbit IgG and anti-LINGO polyclonal antibody via a mini-osmotic pump, respectively. At 3 and 28 days after the operation, the T8～10 segments of the spinal cord were harvested to prepare cryosections, and immunofluorescence staining was used to analyze the penetration of LINGO-1 polyclonal antibody into the spinal cord tissue and its specific binding to LINGO-1 molecules. Results In LINGO-1 antibody group, the presence of rabbit antibodies was detected at the injured sites of the spinal cord at 3 and 28 days after the operation. The mean immunofluorescence density was significantly lower in L1NGO-1 antibody group than in rabbit IgG group at 3 days after the operation (P<0.05). In rabbit IgG group, the mean immunofluorescence density for LINGO-1 in the crysections pre-treated with LINGO-1 polyclonal antibody was significantly lower than that in sections pre-treated with rabbit IgG(P<0.05). Conclusion Locally administered LINGO-1 polyclonal antibody can penetrate into the injured sites in the spinal cord in a wide time window and recognizes LINGO-1 molecule specifically, suggesting the feasibility of passive immunotherapy for spinal cord injury.

Objective: To investigate the origin of infection and risk factors of a case with SARS-CoV-2 infection associated with overseas countries in the Ningbo-Zhoushan Port, Zhejiang Province, so as to provide the evidence for improving the COVID-19 control measures at ports. Methods: Ningbo Center for Disease Control and Prevention ( CDC ) and Beilun CDC conducted case finding and epidemiological surveys immediately after being informed. The general information, history of vaccination and the travel during the latest 14 days were collected from the positive case, and all close contacts were tracked. Saliva samples were collected for SARS-CoV-2 nucleic acid testing and whole-genome sequencing, and the sequencing results were aligned with the GISAID's EpiCoV database. The origin of infection and transmission route of the positive case was investigated. Results: A case was identified positive for SARS-CoV-2 nucleic acid during company M's routine screening in the Ningbo-Zhoushan Port on August 10, 2021, and was confirmed positive for SARS-CoV-2 nucleic acid by Beilun CDC and Ningbo CDC on August 11. Whole-genome sequencing showed SARS-CoV-2 B.1.617.2 ( Delta ) variant, which shared the highest homology with the virus sequence uploaded by Russia on June, 2021 ( Russia/MOW-RII-MH27356S/2021 ). The case was a bundling worker for overseas container ships, and reported communicated with foreign boatmen and contacted materials without protected interventions on the SINOKOR AKITA Container Ship between August 4 and 5, 2021. This ship anchored at Vladivostok, Russia from July 27 to 29, anchored at Ningbo Harbor on August 4, and departed on August 5. Then, 11 boatmen from this ship were tested positive for SARS-CoV-2 nucleic acid on August 8. One asymptomatic case was reported in this epidemic; 254 close contacts and 617 secondary close contacts were identified, and all were tested negative for SARS-CoV-2 nucleic acid. No new cases with SARS-CoV-2 infections were detected until August 25, 2021, and the emergency response was therefore terminated. Conclusions The infection was a sporadic COVID-19 epidemic associated with overseas countries, which was caused by Delta variant infection through contacts with foreign boatmen or materials by a bundling worker in Ningbo-Zhoushan Port; fortunately, no epidemic spread occurred. Intensified closed-loop management and increased frequency of SARS-CoV-2 nucleic acid test among high-risk populations, and improving the precision and rapid emergency treatment of COVID-19 epidemics are required for the containment of COVID-19 at ports.

Background RhoA/ Rho kinase (ROCK) pathway is involved in pulmonary arterial hypertension (PAH) and pulmonary artery smooth muscle cell (PASMC) proliferation. Inhibition of ROCK has been proposed as a treatment for PAH. But the mechanism of RhoA/ROCK pathway and its downstream signaling in proliferation of human PASMCs is unclear. We investigated the effect of fasudil, a selective ROCK inhibitor, on platelet-derived growth factor (PDGF) induced human PASMC proliferation, and the possible association between RhoA/ROCK and extracellular signal-regulated kinase (ERK),p27KiP1.Methods Human PASMCs were cultured with the stimulation of 10 ng/ml PDGF, and different concentrations of fasudil were added before the addition of mitogen. Cell viability and cell cycle were determined with MTT and flow cytometry respectively. ROCK activity, ERK activity and protein expression of proliferating cell nuclear angigen (PCNA) and p27Kip1 were measured by immunoblotting.Results By MTT assay, PDGF significantly increased the OD value that represented human PASMC proliferation, and pretreatment with fasudil significantly reversed this effect in a dose-dependent manner. After PDGF stimulation, the percentage of cells in S phase increased dramatically from 15.6％ to 24.3％, while the percentage in G0/G1 phase was reduced from 80.6％ to 59％. And pretreatment with fasudil reversed the cell cycle effect of PDGF significantly in a dose-dependent manner. PDGF markedly induced ROCK activity and ERK activity with a peak at 15 minutes, which were significantly inhibited by fasudil. In addition, fasudil significantly inhibited PDGF-induced PCNA expression and fasudil also upregulated p27Kip1 expression in human PASMCs, which decreased after PDGF stimulation.Conclusion RhoA/ROCK is vital for PDFG-induced human PASMC proliferation, and fasudil effectively inhibited PDGF-induced human PASMC proliferation by up-regulation of p27Kip1, which may be associated with inhibition of ERK activity.

Objective To investigate the oncolytic effect of E1B mutant adenovirus (d11520) on human malignant gliomas. Methods Ad-βgal vector was used to investigate the infectibility of dl1520.U251,Hep3B (positive control) and T24 (negative control) cell lines were infected with dl1520respectively at 50,5,0.5,0.005 and 0 pfu of multiplicity of infection (MOI).The replication efficiency of d11520 in host cells was assessed by plaque assay.The cytopathic effect (CPE) was assessed by crystal violet staining in a panel of tumor cells. Results Crystal violet staining showed the Hep3B cell line was the most sensitive to dl1520 and had the fastest CPE,followed by the U251 cell line,while the T24cell line had no CEP.The replication and infection rates ofdl1520 in the U251 cell line were lower than in the Hep3B cell line but significantly higher than in the T24 cell line (P＜0.05). Conclusion The E1B mutant adenovirus (dl1520) has a significant oncolytic effect on human malignant gliomas.

AIM To establish the quality standard for Weigela japonica Thunb.var.sinica (Rehd.) Bailey (W.j.).METHODS TLC was adopted in this medicinal material's qualitative identification after morphological identification and microscopic identification.The contents of water,total ash,acid-insoluble ash and extracts were detected according to Chinese Pharmacopoeia methods.Then the contents of scopoletin and total coumarins were determined by HPLC and UV,respectively.RESULTS The morphologies and microscopic characters of W.j.could be distinguished from other same generic plants.The clear TLC spot displayed good resolution.The contents of water,total ash,acid-insoluble ash,water-soluble extract and acid-soluble extract were no more than 12.0％,no more than 2.0％,no more than 0.5％,no less than 5.0％ and no less than 4.5％,respectively.Scopoletin and total coumarins showed good linear relationships within the ranges of 1.25-40.0 μg/mL(r =0.999 7)and 2.0-64.0 μg/mL (r =0.999 9),whose average recoveries were 98.19％ (RSD =0.90％) and 99.21％ (RSD =2.5％),respectively.CONCLUSION This accurate and reliable method can be used for the quality control of W.j..

Objective To investigate the value of the cardiac CT examination for decision making in middle-aged and elderly patients before planned transcatheter atrial septal defect (ASD) closure. Methods Cardiac CT was performed in 63 adult patients [18 males, aged from 50 to 77 years, mean age (56. 87 ±5.79) years]with ASD before planned transcatheter ASD closure. Coronary CT angiography was made for detection of associated cardiovascular diseases, followed by 3D reconstruction of ASD for determination of the defect size in the GE-workstation, results were compared between transthoracic echocardiography measurement, CT measurement, and atrial septal defect occluder waist diameter. Results Cardiac CT identified additional cardiovascular diseases in 14 patients and decision making was changed based on cardiac CT results. Coronary artery stenosis was detected in 8 patients by cardiac CT, and proved by coronary angiography, and all of them were given comprehensive management: percutaneous coronary intervention and thanscatheter ASD closure were successively performed in 2 cases, and 1 case was referred to surgery for both coronary artery bypass graft and surgical ASD repair, and 5 patients were given pharmacological management for coronary artery disease besides thanscatheter ASD closure. Cardiac CT identified large ASD with insufficient rim tissue in 2 cases and transcatheter closures were abandoned.Cardiac CT screened out 1 case from those with insufficient posterior inferior rim by transthoracic echocardiography, and transcatheter ASD closure was successfully pedormed. Cardiac CT ruled out ASD in 1patient. In addition, cardiac CT detected 1 partial abnormalous pulmonary vein connection and 1 ductus arteriosus in this cohort. A correlation on ASD measurements was found between CT size and TTE size(r =0. 80,P ＜ 0. 01 ; Y =0. 84X + 8. 85, R2 =0. 63, P ＜ 0. 05 ), and between ASO size and CT size ( r =0. 92,P ＜ 0. 01 ; Y =0. 93X + 4. 78, R2 =0. 84,P ＜ 0. 05 ). Conclusion In middle-aged and elderly patients with ASD for possible transcatheter closure, cardiac CT is valuable on determing ASD size and morphology and could provide incremental information for optimizing clinical management for ASD patients.

Background The non-operation treatment of intra-abdominal trauma guided contrast enhanced ultrasound (CEUS) is one of the hottest research topic. Gelatin/thrombin/calcium (GTC) was developed as a novel haemostatic agent for non-operable intra-abdominal trauma. We hypothesized that GTC can achieve haemostasis (without the use of pressure)within a short time in a large wound model by percutaneous injection under CEUS guidance.Methods Forty Wister rats received large liver injuries by haemostatic clamp and were randomly divided into four groups, according to the haemostatic agent used. These included normal saline (NS) group A, lyophilising thrombin powder (LTP) group B, GTC group C, and absorbable α-cyanoacrylate (ACNA) group D. Each injury site was treated with one of the above materials and total bleeding time was recorded. All liver wounds were evaluated using CEUS at three periods: pre-injury, injury and post-treatment. The liver wounds were also evaluated by histology 3, 6, and 9 days after injury and the extents of abdominal adhesions were recorded.Results The sensitivity of CEUS (100%) in detecting blunt traumatic liver lesions was significantly higher than conventional ultrasound (42.5%). Bleeding times at the injury site in the GTC group C ((129.3±14.0) seconds) and ACNA group D ((5.2±1.0) seconds) were significantly shorter than those in the NS group A ((369.5±48.8) seconds, P ＜0.01) and LTP group B ((324.7±52.22) seconds, P ＜0.01). The LTP group B showed no significant difference compared with the NS group A. Gross examination of liver tissue revealed that there were fewer intra-abdominal adhesions in the GTC group C (10%) than in the ACNA group D (100%). Histopathologic examination showed that GTC was completely absorbed after nine days.Conclusions GTC, delivered by percutaneous injection under CEUS, may achieve haemostasis (without the use of pressure) within a short time in a large wound model. GTC is absorbable and may prevent intra-abdominal adhesions.Therefore, it may be the optimal choice for first aid treatment of large abdominal wounds in the setting of blunt trauma.

Objective To determine the prevalence and to identify risk factors of peri-procedure electrical storm (ES) in patients with acute myocardial infarction (AMI) underwent emergency percutaneous coronary intervention (PCI).Methods The clinical data of 228 AMI patients underwent emergency PCI were retrospectively analyzed and patients were divided into ES group (n = 39) and non-ES (n = 189) group.ES was referred to spontaneous ventricular tachycardia or ventricular fibrillation occurring twice or more within 24 h and requiring emergency treatment including anti-arrhythm medicine and/or cardioversion or defibrillation.Results ES was diagnosed in 39 out of 228 patients (17.1%) during peri-procedure stage.The incidence of ES in patients with various infarct related arteries (IRA) was as follows:55.6% with left main artery (LM),23.7% with right coronary artery (RCA),12.4% with anterior descending branch (LAD) and 0 with left circumflex artery (LCX).Older age,lager diameter of IRA,higher concentration of CK-MB and cTnT,higher incidence of reporfusion arrhythmia (RA),lower grade of TIMI after PCI and higher mortality were associated with increased risks of ES (The P value was 0.043,0.012,0.036,0.018,0.001,0.049,respectively).Gender,systolic pressure,diastolic pressure,random blood glucose level,white blood count and concentration of hs-CRP were similar between ES and non-ES patients.Logistic analysis showed that the diameter of IRA (OR 2.381,95% CI 1.127-5.028,P = 0.023),TIMI grade of IRA after PCI (OR 4.744,95% CI 1.773-12.691,P = 0.002) and RA (OR 12.680,95% CI 4.360-36.879,P =0.000)were the independent risk factors of per-procedure ES in AMI patients underwent emergency PCI.Conclusions The AMI patients with LM as IRA had the highest incidence of ES during emergency PCI and the diameter of IRA,TIMI grade of IRA after PCI and RA were independent risk factors for the development of ES during peri-PCI stage.

Objective To analyze the usage of structural fat emulsion and medium/long chain fat emulsion, so as to provide reference for promoting its rational use. Methods The demographic data of structural fat emulsion and medium/long chain fat emulsion were sampled from national medical insurance for cities and towns in 2016, demographic characteristics and disease constitution of two fat emulsion patients were statistically analyzed. Results There are statistical differences in the type of insurance, regional distribution and urban grade of the two kinds of drugs used in the structural fat emulsion and medium/long chain fat emulsion.The operation patients are far higher than the non operative group. Structural fat emulsion and medium/long chain fat emulsion were used in the surgical group, and there were statistical differences in the regional, urban and urban levels. The two type of fat emulsion is mainly used for cancer and digestive system diseases. Conclusion The overall usage of structural fat emulsion and medium/long chain fat emulsion is reasonable. It is necessary to further strengthen the rational evaluation of its clinical use.