BACKGROUND:Adenovirus, expressing within a limited period, can limit the expression time and amount of target genes that is not conducive to ongoing experiments. Here, we select adenovirus as vectors for genetic recombination with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (Trail) gene fragment.
OBJECTIVE:To explore the construction of recombinant lentiviral vector carrying Trail in rats
METHODS:The Trail gene was obtained:according to GenBank in rat Trail gene sequence (NM_145681.1), we designed the gene specific primers of Trail-Age I-F and Trail-AgeI-R, and used AgeI as enzyme cutting site. PCR was applied to amplify Trail gene from rat cDNA Library and construct recombinant plasmids after cutting Trail gene to be cloned into expression vector GV218 by AgeI. Recombinant plasmids were transfected into 293T cells by Lipofeetamine2000 encapsulated recombinant plasmid and auxiliary packaging carrier. The Trail protein of lentiviral plasmids was expressed. Fol owing virus col ection, we identified virus titer and extracted protein from cells to detect Trail expression by western blot assay.
RESULTS AND CONCLUSION:Screened positive Escherichia coli DH5a competent cells were sequenced with 861 bp, which was consistent with Trail nucleotide sequence in GenBank. After transfection 2 days, virus liquid was col ected and confirmed as recombinant plasmid including Trail gene by PCR and Trail proteins expressed in 293T cells by western blot assay. Hole dilution method and real-time fluorescent quantitative PCR determination showed that the virus titer was 2×109 TU/mL. In this study, recombinant lentiviral vector carrying Trail is successful y constructed by homologous recombination in Escherichia coli.

Objective To explore the effect of thyroid hormone on bone metabolism.Methods The thyroid in the rabbits had been cut off and the physics and histology of the bone in the rabbits were observed.The therapeutic effect of exterofection thyroid hormone was also observed.Results The lacking or reducing of thyroid hormone could lead to osteoporosis.The substitutive treatment of T_4 could partially inhibit the oesteopenia.Conclusion When the T_3 level equals to the level of T_4,the oesteopenia of the rabbits with thyroid being cut off may be caused by the lack of calcitonin.

Objective To investigate the physicochemica l properties and immunobiological activity of a polysaccharide (RAP-B-1) from stems of Rubus amabilis. Methods The crude polysaccharide (RAP) was obtained successively by boiling, ethanol precipitating and dialyzing. RAP was isolated with DEAE-cellulose and Sephadex G-100 to obtain a polysaccharide RAP-B-1. The physicochemical properties of RAP-B-1 were studied by hydrolysis, periodate oxidation, Smith degradation and methylation, CE, IR, NMR and GC-MS. The immunobiological activities were estimated by the proliferative activity of spleen lymphocytes and phagocytic activity of peritoneal macrophages in mice. Results The molecular weight of RAP-B-1 was 4.80×104 with specific optical rotation value [α] 20D+68.3 (c=1,H2O), and was composed of eight monosaccharides. The molar ratios were as Xyl: Ara: Glc: Rha:Gal: Man: GlcA: GalA = 1.0:6.9:0.8:1.1:6.9:0.3:0.5:3.3. RAP-B-1 was an arabinogalactan. The linkages of arabinose were →1) Ara (2,3→,→1) Ara(5→and→1) Ara, and the linkages of galactose were→1) Gal(4→,→1) Gal(6→and→1) Gal. RAP-B-1 could improve the proliferative activity of spleen T cells(P<0.05) and booste phagocytic activity of peritoneal macrophages at 50μg/ml concentration(P<0.01). Conclusion RAP-B-1 is an arabinogalactan and has immunobiological activity.

Objective The hepatitis B virus (HBV) core protein assembly inhibitors GLS4JHS could destroy HBV capsid assembly and the formation of non-capsid polymer structure.The aim of this study is to explore the mechanisms of GLS4JHS in inhibiting HBV replication.Methods HepAD38 cells was used as the study model.TaqMan real-time polymerase chain reaction (PCR) and quantitative real-time PCR with specific primers were used to measure the change in pregenomic RNA (pgRNA) and covalently closed circular DNA (cccDNA) levels under different concentrations.ChIP assay in HepAD38 cells was used to assess the recruitment of HBV core protein and histone modifications.Results The amount of cccDNA and pgRNA decreased with the increasing GLS4JHS concentrations.After the drug concentrations reached 400 nmol/L, cccDNA and pgRNA declined by 94% and 84%, respectively.Both HBV core protein occupancy on the cccDNA and cccDNA-bound H3 histone acetylation were reduced by GLS4JHS.Conclusions GLS4JHS decreases transcriptional activity of cccDNA and reduces pgRNA production by inhibiting cccDNA minichromosome bound to HBV core protein and acetylated histone H3, which results in HBV DNA formation.

In November 4, 2016, a 1 year and 3 months old male patient with face and neck scald complicated with severe scald of oropharynx was admitted to Guangzhou Red Cross Hospital 1 hour after injury. The child developed upper respiratory tract obstruction 2 hours after injury, therefore tracheotomy and intubation were performed immediately to establish an artificial airway, and symptomatic treatments such as anti-infection, fluid replacement, and dressing change were conducted. On the 10th day after injury, the child had difficult breathing during the test tube blocking before extubation, and it was difficult to extubate. Symptomatic treatments such as ventilator assisted ventilation and strengthened anti-infection were continued. On the 17th day after injury, extubation plan was adjusted. Thirty minutes before extubation, phenobarbital was injected intramuscularly for sedation, and atropine was used to reduce airway secretions, after which extubation was successful. After 21 days of treatment, the child was cured and discharged. In the treatment of this case, high attention was paid to the important influence of children′s mental factors among causes of difficult extubation, which provided a reference for clinical treatment of extubation in children with tracheal tube after tracheotomy.

The arm span is the sum of the length of the arm and the width of the chest.It is highly correlated with height and can be used as an accurate and reliable alternative measure to estimate height.The arm span/height ratio reflects the relationship between long bones and the trunk and can be used to evaluate the body proportion.A correct understanding of the development patterns and characteristics of arm span can help pediatricians to find the deviation of body proportion in time, which is of great significance to the etiological analysis of short stature.

Objective To determine the scores of patients with a confirmed diagnosis of drug-induced liver injury(DILI)using Roussel Uclaf Causality Assessment Method(RUCAM),Maria&Victorino assessment scale,and Revised Electronic Causality Assessment Method(RECAM),to compare the accuracy of the three scales in diagnosis,and to investigate their clinical significance in the diagnosis of DILI.Methods A total of 98 patients with a confirmed diagnosis of DILI who were hospitalized in Peking University First Hospital from January 2011 to December 2022 were enrolled,with liver biopsy results supporting DILI and a clear history of medication.Clinical data were collected from all subjects,and the above causality assessment scales were used for scoring.The chi-square test was used to analyze the diagnostic accuracy of the causality assessment scales,and the weighted kappa coefficient was used to analyze the consistency between the three scales.Results For all patients with DILI enrolled,RECAM had the highest accuracy,with a significant difference compared with RUCAM(χ2=5.667,P=0.017).RUCAM and RECAM had moderate consistency in diagnosis(κw=0.469),while RECAM and Maria&Victorino scale had poor consistency(κw=0.156).For the patients with acute DILI,RECAM,RUCAM,and Maria&Victorino scales had a diagnostic inconsistency rate of 3.7%,11.1%,and 42.6%,respectively;for the patients with hepatocellular type DILI,the three scales of a diagnostic inconsistency rate of 8.9%,21.4%,and 62.5%,respectively;for the patients with cholestasis type or mixed type DILI,the three scales of a diagnostic inconsistency rate of 10.0%,22.5%,and 47.5%,respectively.Conclusion The use of RECAM and RUCAM scales in acute DILI can improve diagnostic rate,and for hepatocellular type DILI and DILI with the clinical manifestation of cholestasis(cholestasis type DILI and mixed type DILI),the use of RECAM and RUCAM scales can also improve diagnostic rate.The selection of causality assessment scales with a relatively high accuracy based on the course and clinical classification of the disease may help to further improve clinical diagnostic rate.

Objective:To investigate the effect of radiofrequency radiation (RF) from 5G mobile phone communication frequency bands (3.5 GHz and 4.9 GHz) on the permeability of the blood-brain barrier (BBB) in mice.Methods:A total of 24 healthy adult male C57BL/6 mice (6-8 weeks old) were randomly divided into Sham, 3.5 GHz RF and 4.9 GHz RF groups, and 8 mice in each group. Mice in the RF groups were systemically exposed to 5G cell phone radiation for consecutive 35 d(1 h/d) with 50 W/m 2 power density. The BBB permeability of mice was detected by Evans Blue (EB) fluorescence experiment. The expression levels of the BBB tight junction-related proteins (ZO-1, occludin and claudin-11) and the gap junction-related protein Connexin 43 were determined by Western blot. Results:The number of spots, fluorescence intensity and comprehensive score of EB were significantly increased in 3.5 GHz RF group and 4.9 GHz RF group compared with the Sham group ( t=12.98, 17.82, P<0.001). Compared with the Sham group, the content of S100B in mouse serum was significantly increased in 3.5 GHz RF group and 4.9 GHz RF group ( t=19.34, 14.68, P<0.001). The BBB permeability was increased in the RF group. The expression level of occludin protein was significantly reduced in the 3.5 GHz RF group ( t=-3.13, P<0.05), and this decrease was much profound in the 4.9 GHz RF group ( t=-6.55, P<0.01). But the protein levels of ZO-1, Claudin-11 and Connexin 43 in the cerebral cortex of the RF groups had no significantly difference in comparison with the Sham group( P>0.05). Conclusions:The continuous exposure of mobile phone RF at 3.5 GHz or 4.9 GHz for 35 d (1 h/d) induces an increase of BBB permeability in the mouse cerebral cortex, perhaps by reducing the expression of occludin protein.

Objective:To explore the effect of task-driven group workshop learning method on teaching satisfaction degree and practical ability in Mongolian medical nursing students.Methods:From September 2018 to January 2020, using convenient sampling method, a total of 38 Mongolian undergraduate nursing students of grade 2016 of Mongolian Medical College of Inner Mongolia Medical University were recruited as control group to receive traditional method, and the teaching practice activities were arranged after the main content was completed. Another 39 students of grade 2017 were recruited as observational group to receive task-driven group workshop learning method. The preparation of teaching practice tasks will run through all stages of teaching. The differences of students′ achievement, teaching satisfaction and autonomous learning ability between the two groups were compared.Results:The grade of observational group was 84.81 ± 3.45, higher than 76.16 ± 3.59 of the control group , which had significant difference ( t=-5.35, P<0.05). The score of teaching satisfaction of observational group was 82.00 ± 11.62, higher than 70.94 ± 6.65 in the control group, which had significant difference ( t=-5.10, P<0.05). The dimensions of information ability, cooperation ability and total score of the Autonomous Learning Ability Scale in observational group scored 39.28 ± 6.46, 24.54 ± 3.45, 98.13 ± 14.58, which were higher than 36.18 ± 5.46, 22.39 ± 3.59, 91.37 ± 11.47 in the control group, which had significant difference ( t=-2.27, -2.67, -2.26, all P<0.05). Conclusions:The task-driven group workshop learning method can improve the study result and the satisfaction of teaching, it can also improve the information ability and cooperation ability in Mongolian medical nursing students, which is worthy of reference in nursing teaching.

Hepatitis B virus core antibodies are specific antibodies produced after viral infection that appear early and last for a long time, and its levels in serum are measured by the double-antigen sandwich chemiluminescent microparticle immunoassay method, which has higher sensitivity and specificity, providing new clinical indicators for hepatitis B patients diagnosis, treatment, and drug withdrawal management. This article reviews the clinical significance and research progress of quantitative hepatitis B core antibody measurement and expounds on its research applications and prospects in clinical practice.