Preparation and Biodistribution of 131I-labeled Hepatoma Nucleic Acid Nanotrain / 中山大学学报(医学科学版)

ObjectiveTo construct 131Ⅰ-labeled hepatoma nucleic acid nanotrain and to explore its feasibility as a new nuclide carrier targeting hepatoma. MethodsThree short nucleic acid chains self-assembled to a long nucleic acid chain after being annealed， and 131Ⅰ-NT was obtained by radioiodine labeling using chloramine T method. The labeling efficiency and radiochemical purity of the nanoparticles were measured by paper chromatography. The stability of the labeled products in vitro at different temperatures and different storage solvents was detected. The specific uptake of nanoparticles by hepatocellular carcinoma cells was observed by laser confocal microscopy， and the radioactive uptake ratio of 131Ⅰ-NT combined with human hepatocellular carcinoma cell HepG2 and normal hepatocyte L02 was measured. The biodistribution of 131Ⅰ-NT was obtained through injecting 131Ⅰ-NT into HepG2 tumor-bearing mice via tail vein. ResultsThe labeling rate of 131Ⅰ-NT was （93.05±0.74） %， and the radiochemical purity post purification was （98.35±0.32） %. Its radiochemical purity in PBS and pure serum at 4℃ for 24 h was （92.77±0.04） % and （89.43±0.2） %， respectively. The radioactivity uptake rate of HepG2 cells was higher than that of L02 cells after 131Ⅰ-NT was incubated with two kinds of cells for 2 h significantly. After injection of 131Ⅰ-NT through tail vein， the radioactive uptake per gram of tumor tissue were （4.9±0.55）%ID/g， （10.12±0.32）%ID/g and （4.25±0.31）%ID/g at 30 min， 1 h and 2 h， respectively. The T/M ratio was 7.33±2.04， 36.54±12.72 and 44.93±7.90 respectively. ConclusionsThe 131Ⅰ-labeled long chain nucleic acid nanotrain was constructed successfully， which possesses relatively high stability in vitro ， and high targeting ability to HepG2 cells in vitro and HepG2 tumor-bearing mouse model. Our study demonstrated that 131Ⅰ-NT may be a potential radionuclide carrier targeting human liver cancer， which provides a new idea for the targeted radionuclide diagnosis and treatment of hepatocellular carcinoma.

Changes of corneal parameters in different areas after cataract surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To compare the distribution characteristics of axial sagittal front power(ASF), true net power(TNP), total corneal refractive power(TCRP)and the difference in back-front corneal radius ratio(B/F ratio)after cataract surgery.METHODS: A prospective study. A total of 156 patients(156 eyes)with age-related cataract who attend Weifang Eye Hospital for cataract surgery from December 2020 to May 2021 were collected. Pentacam was performed before operation and 3mo after operation to collect ASF, TNP and TCRP on 2, 4 and 6mm diameters rings and areas on the corneal apex and pupil-centered, as well as B/F ratio.RESULTS: 3mo after operation, there was no statistical difference in ASF on the 2mm diameters ring and area centered on the corneal apex compared with preoperative values(all P>0.05), however, the ASF values on the 4 and 6mm diameters rings and areas were significantly different from those before surgery(all P<0.05); There was no statistical difference in ASF on the 2mm diameters rings and areas centered on the pupil compared with preoperative values(all P>0.05). The postoperative values of TNP and TCRP on the 2, 4 and 6mm diameters rings and areas centered on the corneal apex and centered on the pupil were statistically different before surgery(all P<0.05). Preoperative, TCRP values were different between 2mm and 6mm and between 4mm and 6mm on both corneal apex and pupil-centered rings(all P<0.0167), TCRP values were all different between 2mm and 6mm diameters areas on corneal apex and pupil-centered(all P<0.0167); 3mo after operation, TCRP values were different on corneal apex and pupil-centered rings between 2mm and 6mm and between 4mm and 6mm diameters(all P<0.0167). While TCRP values on the corneal apex and pupil-centered areas were only different between 2mm and 6mm diameters(all P<0.0167). The preoperative B/F ratio of patients was 81.79%±1.87%, and the postoperative B/ F ratio of patients was 80.68%±2.23%(P<0.001).CONCLUSION: Corneal parameters of different diameters of rings and areas centered on the corneal apex and pupil before and after cataract surgery may change and differ, which should be taken into account when selecting the K value for intraocular lens calculation and individualizing the selection of IOLs based on corneal characteristics.

Polyphyllin I promotes cell death via suppressing UPR-mediated CHOP ubiquitination and degradation in non-small cell lung cancer / 中国天然药物

Polyphyllin I (PPI) purified from Polyphyllarhizomes displays puissant cytotoxicity in many kinds of cancers. Several researches investigated its anti-cancer activity. But novel mechanisms are still worth investigation. This study aimed to explore PPI-induced endoplasmic reticulum (ER) stress as well as the underlying mechanism in non-small cell lung cancer (NSCLC). Cell viability or colony-forming was detected by MTT or crystal violet respectively. Cell cycle, apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential were assessed by flow cytometry. Gene and protein levels were evaluated by qRT-PCR and immunoblotting respectively. Protein interaction was determined by immunoprecipitation or immunofluorescence assay. Gene overexpression or silencing was carried out by transient transfection with plasmids or small interfering RNAs. The Cancer Genome Atlas (TCGA) database was used for Gene Set Enrichment Analysis (GSEA), survival analysis, gene expression statistics or pathway enrichment assay. PPI inhibited the propagation of NSCLC cells, increased non-viable apoptotic cells, arrested cell cycle at G2/M phase, induced ROS levels but failed to decrease mitochondrial membrane potential. High levels of GRP78 indicates poor prognosis in NSCLC patients. PPI selectively suppressed unfolded protein response (UPR)-induced GRP78 expression, subsequently protected CHOP from GRP78-mediated ubiquitination and degradation. We demonstrated that the natural product PPI, obtained from traditional herbal medicine, deserves for further study as a valuable candidate for lead compound in the chemotherapy of NSCLC.

Neuroprotective effect of luteolin-7-O-β-D-glucuronide in a rat model offocal cerebral ischemia / 中国药理学与毒理学杂志

OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.

High fat diet-induced elevation of blood glucose impairs the microvascular function of testis in mice / 基础医学与临床

Objective To observe the effects of high fat dietinduced elevation of blood glucose on the microvascular function of testis and male reproduction in C57BL/6 mice. Methods A total of 40 male C57BL/6 mice were randomly divided into control group and high fat diet (HFD) group (n =20). The mice in HFD group were fed with high fat diet for 20 weeks. Blood glucose and body weight were measured weekly. The permeability of bloodtestis barrier was evaluated by intraperitoneal injection of Evans blue. The blood flow of testicular microcircu-lation and the frequency and amplitude of microvascular vasomotion were detected by laser Doppler blood flow ima-ging system. The morphology of testicular tissue was observed by HE staining. The expressions of platelet- endothelial cell adhesion molecule-1 (PECAM-1/CD31) in testicular microvascular endothelial cells and prolifera-ting cell nuclear antigen (PCNA) in spermatogenic cells were detected by immunohistochemistry. The apoptosis of spermatogenic cells was observed by TUNEL staining. Results The body weight and blood glucose of HFD group were significantly higher than those in control group (P＜0.01). Evans blue staining showed that the integrity of blood-testis barrier of HFD group was damaged, and increased permeability was observed in seminiferous tubules. In HFD group, the mean blood flow of testis and the frequency and amplitude of microvascular vasomotion were sig-nificantly lower than those in control group (P＜0.01). The number of spermatogenic epithelial cells and the thickness of seminiferous epithelium decreased. The expressions of CD31 in microvascular endothelial cells and PCNA in spermatogenic cells were significantly lower in HFD group than those in control group (P＜0.01). The apoptosis level of spermatogenic cells was higher than that in the control group (P <0.01). Conclusions Increased blood glucose level induced by high fat diet in mice can impair the testicular microvasculature and damage the integrity of blood-testis barrier and injure the structure of seminiferous epithelium in mice.

Investigation for Current Status of Tachyarrhythmia Treatment in Emergency Patients in Beijing / 中国循环杂志

Objective: To investigate the current status of tachyarrhythmia treatment and outcomes in emergency patients. Methods: A total of 250 tachyarrhythmia patients treated in emergency departments from 1 tertiary hospital, 1 secondary hospital and 1 cardiovascular hospital in Beijing were enrolled. The baseline condition, type of tachyarrhythmia such as atrial fibrillation (AF)/atrial flutter, intravenous medication, prognosis and the choice of anticoagulation therapy were collected and statistically analyzed. Results: The mean age of patients was 64 years including 123/250 (49.2%) female. Common previous histories included 135 (54.0%) cases of hypertension, 93 (37.2%) cases of CAD, 29 (11.6%) cases of stroke/TIA, 29 (11.6%) cases of valvular heart disease and 18 (7.2%) cases of cardiomyopathy; 136 (54.4%) cases combined with organic heart disease. 123 (49.2) cases had previous atrial flutter/AF. The most common type of tachyarrhythmia was AF (172 cases, 68.8%) and the rest in turn, were supra-ventricular tachycardia (41 cases, 16.4%), ventricular tachycardia (23 cases, 9.2%), atrial flutter (21 cases, 8.4%), atrial tachycardia (12 cases, 4.6%) and premature ventricular contraction (5 cases, 2%). The most commonly used drug for treating supra-ventricular tachycardia was propafenone (151 cases, 60.4%), commonly used drug for treating supra-ventricular tachycardia and ventricular arrhythmia was amiodarone. With proper treatment, arrhythmia was stopped in 56% (140 cases), improved in 40% (100 cases) and unchanged or died in 10% (25 cases). 107 non-valvular AF/atrial flutter patients received low molecular heparin for anticoagulation therapy and only 25 patients continued oral anticoagulant therapy after discharge.Conclusion: In our research, most emergency patients were treated consistently with current guidelines. The choice of medication and anticoagulation therapy in AF patients should be improved.

Establishment of Automation System for Detection of Alcohol in Blood / 法医学杂志

OBJECTIVES@#To establish an automation system for detection of alcohol content in blood.@*METHODS@#The determination was performed by automated workstation of extraction-headspace gas chromatography （HS-GC）. The blood collection with negative pressure, sealing time of headspace bottle and sample needle were checked and optimized in the abstraction of automation system. The automatic sampling was compared with the manual sampling.@*RESULTS@#The quantitative data obtained by the automated workstation of extraction-HS-GC for alcohol was stable. The relative differences of two parallel samples were less than 5%. The automated extraction was superior to the manual extraction. A good linear relationship was obtained at the alcohol concentration range of 0.1-3.0 mg/mL （r≥0.999） with good repeatability.@*CONCLUSIONS@#The method is simple and quick, with more standard experiment process and accurate experimental data. It eliminates the error from the experimenter and has good repeatability, which can be applied to the qualitative and quantitative detections of alcohol in blood.

Isolation, identification, and pathogenicity of neospora cani-num China Yanbian strain

The aim of the study was to provide a point of reference to study the Neospora caninuminfections in China. Genome DNA was extracted from the brains of aborted fetuses and specific PCR was performed with N. caninum Nc5-targeted specific primers. Fetal bovine brain tissues were homogenized and continuously cultured in Vero cells with double antibodies. The medium was replaced at 2-d intervals and the state of cells was observed. A 608 bp Nc5 gene band was detected by PCR amplification. After sequencing, the sequence of the sample shared 99.5% homology with GenBank [AF061249]. Brain homogenates were continuously cultured in Vero cells for 34 d and N. caninum was found. The results of IFAT and Nc5 gene-based PCR detection were N. caninum-positive, and the parasite was tentatively named N. caninum China Yanbian strain. BABL/c mice were inoculated with the separated parasites and showed clinical symptoms of ataxia and limb paralysis after 12 d. Only 3 mice survived. The blood of dying mice and the hearts, livers, spleens, lungs, kidneys, and brains of dead mice were collected aseptically. The Nc5 gene-based PCR showed that N. caninum may exist in brains, livers, and spleen. Based on immunohistochemical observations, we showed that N. caninumtachyzoites existed in the brains and livers. We have successfully isolated bovine-specific N. caninum strain from brain tissues of aborted cattle in the China Yanbian region. This isolated strain has a strong infectious ability towards BABL/c

Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Thyroid peroxidase (TPO) is an important autoantigen in Hashimoto's thyroiditis (HT), and almost all epitopes are located in TPO ectodomain. The glycosylation of TPO might contribute to breaking self-tolerance, therefore, purified glycosylated recombinant TPO ectodomain is prerequisite of elucidating its role in the pathogenesis of HT. The aim of our study was to investigate whether the glycosylation has influence on the antigenic determinants of recombinant TPO.</p><p><b>METHODS</b>Bac-to-Bac baculovirus expression system was used to generate recombinant human TPO ectodomain. The antigenicity was analyzed by antigen specific enzyme-linked immunosorbant assays (ELISAs). The glycosylation of recombinant human TPO ectodomain of High Five insect cell origin was detected by lectin-ELISAs.</p><p><b>RESULTS</b>TPO ectodomain was recovered from the culture media as a soluble protein, and it was fused with a hexahistidine tag which allowed purification by nickel-affinity chromatography. The recombinant TPO ectodomain could be recognized by all the 54 HT patients and three TPO monoclonal antibodies. Fucose, sialic acid and galactose were all detected on the recombinant TPO ectodomain. Sera TPOAb binding decreased slightly after non-specific deglycosylation of TPO by periodic acid.</p><p><b>CONCLUSIONS</b>High Five insect cells derived recombinant human TPO ectodomain had N-glycosylation sites, which might have little effect on recognition by serum TPOAb.</p>

Generation and Immunity Testing of a Recombinant Adenovirus Expressing NcSRS2-NcGRA7 Fusion Protein of Bovine Neospora caninum

Neospora caninum is the etiologic agent of bovine neosporosis, which affects the reproductive performance of cattle worldwide. The transmembrane protein, NcSRS2, and dense-granule protein, NcGRA7, were identified as protective antigens based on their ability to induce significant protective immune responses in murine neosporosis models. In the current study, NcSRS2 and NcGRA7 genes were spliced by overlap-extension PCR in a recombinant adenovirus termed Ad5-NcSRS2-NcGRA 7, expressing the NcSRS2-NcGRA7 gene, and the efficacy was evaluated in mice. The results showed that the titer of the recombinant adenovirus was 10(9)TCID50/ml. Three weeks post-boost immunization (w.p.b.i.), the IgG antibody titer in sera was as high as 1:4,096. IFN-gamma and IL-4 levels were significantly different from the control group (P<0.01). This research established a solid foundation for the development of a recombinant adenovirus vaccine against bovine N. caninum.