Objective:To create a large human phage antibody library from which easy to get diabody and select human antibody clones.Methods:VL and VH genes were amplified by RT-PCR from RNA which came from normal adult peripheral blood and new-born cord blood lymphocytes. VL and VH genes were reamplified to add a region of overlap in the ScFv linker to form ScFv genes in which VH genes were flanked by two non-homologous loxp sites. The ScFv genes were cloned into PDF to obtain a primary library. This primary library was used to infect bacteria Bs1365(expressing cre recombinase) with high multiplicity of infection(MOI) 200∶1. This procedure resulted in a very large phage antibody library through VL and VH recombined by the cre recombinase. Following recombination, phagemide were derived from these bacteria and used to infect bacteria not expressing cre(XL1-Blue) at MOI

A high performance liquid chromatography-tandem mass spectrometric ( HPLC-MS / MS) method coupled with an immunoaffinity clean-up column was successfully developed for determination of aflatoxins (AFB1 , AFB2 , AFG1 , AFG2 , AFM1 and AFM2 ) and zeranols ( α-zeranol, β-zeranol, α-zearalenol,β-zearalenol, zearalanone and zearalenone ). The sample was extracted with methanol-acetonitrile (20∶ 80, V/ V) after enzymatic digestion by β-glucuronidase / sulfatase, and the extraction solution was passed through glassy fiber filter paper and then diluted with phosphate buffer solution (PBS). The reconstituted solution was cleaned up with IAC-AZ immunoaffinity column, and then analyzed by HPLC-MS / MS in multiple reaction monitoring (MRM) mode. The results indicated that the linear detection range was 0. 03-6. 0 μg / L for AFB2 and AFG2 , and 0. 05-20 μg / L for the rest compounds. The correlation coefficients were above 0. 999. The limits of detection (LOD) and limits of quantitation (LOQ) were 0. 01-0. 03 μg / kg and 0. 04-0. 09 μg / kg, respectively. The recoveries of the aflatoxins and zeranols were in the range of 73. 6% -98. 4% at the spiked levels of 0. 5, 1 and 5 μg / kg, and the relative standard deviations (RSDs) were in the range of 1. 9% -11. 2% . The method was proved to be simple and accurate, and suitable for the rapid determination of aflatoxins and zeranols in animal-originated foods.

Objective The purpose of this experiment is to study the in vivo differentiation and myelin formation of rat striatal neural precursor cells after transplantation into homogeneous retina,observe the order of myelination and its influence on the structure of retina,establish an animal model of CNS myelin formation in vivo. Methods Passage cultured striatal neural precursor cells from embryonic Sprague-Dawley rats were transplanted into the vitreous cavity of neonatal rats.In different stages after transplantion,myelin formation in retina was observed under light and electron microscope and analysed with different stained methods.Results Bundles of myelin appeared in parts of retina 4 weeks later.The distribution and morphology of myelined area expanded with prolonged survival time after cell transplantation.Oligodendrocyte wrapped the naked axons and formed normal myelin limited in the nerve fiber layer.Myelination influenced the distribution of local retinal ganglion cells.Conclusion Striatal neural precursor cells could differentiate into oligodendrocytes and formed myelin after transplanted into retina and the naked axons in retina promoted the myelin formation.This model provides a new method to study the myelin formation and myelin-axon interaction in vivo.

Objective To explore the general rules of complications of hemophilia based on the disease network and predict the possible complications of hemophilia patients.Methods The PubMed database was searched for literature related to hemophilia.Disease entities were extracted from the ti-tles and abstracts of the literature using the MetaMap tool.A disease network was constructed based on the co-occurrence relationship of disease pairs,and the overall characteristics,node characteristics,and structural characteristics of the disease network were analyzed.The disease entity network was an-alyzed for association rules to explore the general patterns of hemophilia complications.The link pre-diction algorithm was used to predict potential complications of hemophilia.Results The association network composed of hemophilia and its complications satisfied the characteristics of small-world net-works and evenly distributed cohesive subgroups in terms of network structure.Cohesive subgroup a-nalysis showed that hemophilia complications can be divided into four categories:inherited diseases,blood system diseases,infectious diseases,and chronic diseases.Association rule analysis identified 133 rules with a confidence level ≥0.8,and link prediction further identified many well-documented disease pairs.Conclusion Based on the disease network,hemophilia complication association analy-sis and link prediction can effectively predict potential hemophilia complications,providing decision support for clinical diagnosis and treatment of hemophilia.

Objective To explore the general rules of complications of hemophilia based on the disease network and predict the possible complications of hemophilia patients.Methods The PubMed database was searched for literature related to hemophilia.Disease entities were extracted from the ti-tles and abstracts of the literature using the MetaMap tool.A disease network was constructed based on the co-occurrence relationship of disease pairs,and the overall characteristics,node characteristics,and structural characteristics of the disease network were analyzed.The disease entity network was an-alyzed for association rules to explore the general patterns of hemophilia complications.The link pre-diction algorithm was used to predict potential complications of hemophilia.Results The association network composed of hemophilia and its complications satisfied the characteristics of small-world net-works and evenly distributed cohesive subgroups in terms of network structure.Cohesive subgroup a-nalysis showed that hemophilia complications can be divided into four categories:inherited diseases,blood system diseases,infectious diseases,and chronic diseases.Association rule analysis identified 133 rules with a confidence level ≥0.8,and link prediction further identified many well-documented disease pairs.Conclusion Based on the disease network,hemophilia complication association analy-sis and link prediction can effectively predict potential hemophilia complications,providing decision support for clinical diagnosis and treatment of hemophilia.

Objective To discuss the influence of Tongluo Baoshen Fufang (TBF) and its disassembled formulas on P-IRE1α, promoter of endoplasmic reticulum stress (ERS) and GRP78, molecular chaperone, in diabetic rats with hyperglycemic kidney injury induced by streptozocin (STZ).Methods Male SD rats were randomly divided into normal group and modeling group.The model of diabetes was established by applying intraperitoneal injection of STZ in model group.After successfully modeling, the modeling group was randomly divided into model group, valsartan group(8.93 mg/kg), TBF group(19.43 g/kg), Tongluo Baoshen Buxu group (TBB group,6.92 g/kg) and Tongluo Baoshen Jiedu group (TBJ group,12.5 g/kg).The general conditions of rat daily activities were observed in all groups.The expressions of p-IRE1α and GRP78 were analyzed semi-quantitatively by using immunohistochemistry and IPP software after 6 weeks and 16 weeks.Results P-IRE1α and GRP78 mainly existed in the cytoplasm of proximal and distal renal tubule cells and glomerular cells observed by light microscope.The results of semi-quantitative immunohistochemistry and IPP software showed that the expressions of P-IRE1α and GRP78 increased in model group compared with normal group after 6 weeks and 16 weeks.Compared with model group, the expressions of P-IRE1α and GRP78 decreased in valsartan group, TBF group, TBB group and TBJ group (P<0.05) after 6 weeks and 16 weeks.The comparison among valsartan group, TBF group, TBB group and TBJ group did not show difference(P>0.05).Conclusion The rat model of kidney injury induced by hyperglycemia has ERS activation.TBF and its disassembled formulas can take play a renal protective role through inhibiting ERS.It can be speculated that the central pathogenesis of diabetic hyperglycemic kidney injury is deficiency of the root and excess of the tip in Chinese medicine.

OBJECTIVE To mine the focus and emotional attitude of the public on rare diseases， and to provide decision- making reference for relevant departments to formulate and implement relevant policies， systems and strategies for medical security of rare diseases. METHODS Latent Dirichlet allocation （LDA） topic model and sentiment analysis method were used to analyze the comment text of short videos related to the “nosinasine” medical insurance admission event on the Bilibili video website， mine the theme and sentiment tendency of the text， and put forward relevant strategy suggestions. RESULTS A total of 8 videos with tens of thousands of playback and 7 109 text data were obtained. According to the LDA analysis， online public paid attention to 9 topics related to rare diseases （the price of rare disease drugs， the inclusion of rare disease drugs in medical insurance， the status quo of research and development of rare disease drugs， the scope of medical insurance and the medical security system， the plight of rare disease patients， the prevention and screening of rare diseases， the value of rare disease drugs in medical insurance， the Chinese and western medical methods of rare diseases， and the supply and demand status of rare disease drugs）， which could be summarized into 4 categories according to the objects of concern （rare disease drugs， rare disease medical insurance， rare disease medical and health services and rare disease patient groups）. On the whole， the emotional tendency of the Internet public towards each topic showed a low positive and high negative tendency. CONCLUSIONS The public paid more attention to the price， research and development， supply and demand of rare disease drugs， and was deeply worried about the current medical status of rare diseases in China. To strengthen and improve the medical security for patients with rare diseases， such as actively carrying out public health services for rare diseases， strengthening cooperation in the diagnosis and treatment of rare diseases， and researching rare disease drugs， etc. Chinese medical and health departments can make concerted efforts in medical and health services， drug supply security， medical security and other social security.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.