Objective:To develop non-immunized human phage display library.Methods:The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:The developed phage library is diversity and can be used for selecting humanized scFv.

AIM:To investigate the effect of lipopolysaccharides(LPS)preconditioning on CCl4-induced liver injury and the change of LPS signal transduction.METHODS:The male Wistar rats were divided randomly into liver-injury group,which were injected with CCl4 5 mL/kg first,three days later were injected 0.3 mL 40% CCl4 and 60% olive oil. Animals in LPS preconditioning group were injected with LPS 0.5 mg/kg before the day CCl4 was given. Rats received high fat diet were as liver injury group,and normal control group received normal diet. The lymphocytes infiltrated in the liver tissue were counted. The endotoxin and ALT level in rat plasma,TNF-? content and expressions of TLR4,p38,p-p38,I??,NF-?? in the rat livers were also determined.RESULTS:The lymphocytes in liver slice and ALT level of the plasma in LPS preconditioning group were lower significantly than those in the liver injury group,and the expressions of TLR4,p-p38,NF-?? in the liver were the same. In contrast,the expression of I?? was higher.CONCLUSION:LPS preconditioning relieves obviously CCl4-induced chronic liver injury. The mechanism may be associated with change of signal transduction of LPS,which results in decrease of pre-inflammatory cytokines.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

AIM: To determine the effect of Radix Angelicae Sinensis(RAS) on renal ischemia/reperfusion injury in rabbits and to explore its mechanism. METHODS: Twenty-five rabbits were divided randomly into the sham operated group(Control group), renal ischemia/reperfusion injury group(IR group) and RAS+IR group. At the time point of reperfusion 48 h after renal ischemia 1 h, the renal tissue were observed by electron-microscope and the contents of creatinine(Cr) in serum, tumor necrosis factor-?(TNF-?), interleukin-6(IL-6)and basic fibroblast growth factor(bFGF) in the renal tissue were measured. RESULTS: A remarkably degenerative changes in renal tissue were showed under electronmicroscope in IR group , but the changes in RAS+IR group were slight . The contents of Cr, TNF-? and IL-6 in IR group were higher than those in Control group, these parameters in RAS+IR group were lower than those in IR group, the difference between these groups was significant ( P

To investigate and summarize the characteristics of mass media reports about medical disputs, and to analyze how the reports impact public perception and behaviors, and finally to propose some suggestions. A total of 385 people were conveniently randomizedly sampled to the survey, including 280 citizens in Beijing and 105 netizens. The results showed reporting modes by the mass media were diversified, and their truthfulness was various. Among them, 63.56% people thought the mass media reporting “medical disputs” almost supported the truthfulness, while 23.29% thought the mass media lacked truthfulness, because the format of the mass media reporting was fast speed rather than good quality. A total of 98.63% people thought the mass media has an impact on the public perception and behaviours, where 69.36% supported their positive aspects, while 30.64% showed negative. Based on the views above, it’s found that the mass media lacked right information of medicine. Therefore, in order to make a harmonious condition of doctor-patient relationship, it’s necessary to truly report the news with a scientific view, to build a platform for the information exchanges between hospitals and media, and to strengthen the social supervision and management.

In recent years, for patients with acute ischemic stroke due to large vessel occlusion, intravascular mechanical thrombectomy has become the most recommended treatment method. Determining whether there is ischemic penumbra or not is very important for the selection of patients who are suitable for endovascular mechanical thrombectomy. In addition, the ischemic penumbra also provides guidance for extending the time window of traditional intravenous thrombolytic therapy. This article reviews the imaging evaluation methods of ischemic penumbra.

Background In recent years,some researches had been conducted on the pathologic changes of the secondary injury of perihematoma in animal experiments,but only a few studies had been done on the dynamic pathologic and ultrastructural changes of the perihematoma in ICH patients. The unique contribution of our study is to investigate the dynamic pathologic and ultrastructural changes of the perihematoma in ICH patients and provide significant insights into how the pathophysiology and ultrastructures changed after ICH.Methods The written informed consents were obtained from the ICH patients or their relatives. 30 patients (the supertentorial hemotoma volume＞30 mi and the cerebellar hemotoma volume ＞10 mi) were divided into 8 groups according to the time passed after ICH:＜6 h (6 patients), 6 ～ 12 h (7 patients), 12 ～24 h (5 patients), 24～48 h (3 patients), 48 ～72 h (3 patients), 3 ～4 days group (3 patients), 5 days group (2 patients) and 8 days group ( 1 patient) and subjected to craniotomy for hemotoma evacuation. During the operation for the hemotoma's evacuation, a small amount of tissues that must be removed, which located at 1 cm near the hematoma, were taken as experimental groups; And the same tissues of 7 patients (＜12 h), which were far from the hemotoma on the operational way, were taken as control group. The pathologic and ultrastructral changes were observed.Results The tissues of the control group were almost normal while the damages of the tissues from the experimental groups were slight in ＜6 h groups, more severe after 6h and got to the maximum between 24 ～48 h , recovered gradually after 72 h, became similar to the 6 ～ 12 h group on 5 th day, got better on 8 th day and resembled the 6 h group.Conclusions The injury of the perihematoma occurred in early stage, reached the peak level between 24 and 48 hours after ICH; which was consistent to the clinical nervous functional deficits in the ICH patients.

Objective: To report the clinical features of anti-MDA5 antibody positive juvenile dermatomyositis (JDM) complicated with severe interstitial lung disease (ILD). Methods: The clinical data of three patients, who was admitted to the Department of Rheumatology and Immunology, Children's Hospital of the Capital Institute of Pediatrics from September 2016 to July 2017, with anti-melanoma differentiation associated gene 5 (MDA5) antibody positive JDM complicated with ILD were retrospectively extracted and analyzed. Meanwhile, PubMed database, CNKI, Wanfang database and China Biology Medicine disc (from their establishment to February 2019) with the key words "juvenile dermatomyositis" "interstitial lung disease" , and "anti-MAD5 antibody" both in English and Chinese were searched. Results: There were 2 females and 1 male (P1-P3), aged from 10 years 3 months to13 years 4 months, the time from onset to diagnosis were 2 months, 4 months and 10 months. All presented with rash. One of them had decreased muscle strength, and two had decreased activity tolerance. Creatine kinase was 588, 915 and 74 U/L, and serum ferritin were 1 792, >2 000 and 195.4 μg/L. All three patients had positive anti-MDA5 antibodies. At the time of diagnosis, all of them had ILD, pneumothorax and mediastinal emphysema, but had no respiratory symptoms. All three patients received oral methylprednisolone and cyclophosphamide pulse therapy, while human immunoglobulin was given only to P1 and P2. P1 developed rapid progressive pulmonary interstitial disease (RPILD) and died of respiratory failure after 2 months. While P2 and P3 were followed up for 1 to 2 years, who had complete remission, as anti-MDA5 antibody turned to negative and ILD improved significantly. Ten related reports in literature were retrieved, without reported Chinese cases, and most cases initiated with rash and very likely complicated with arthritis. Some of them were more likely to have ILD rather than muscle weakness. It also showed that Japanese JDM children had higher rate of positive anti-MDA5 antibody than patients from the U.S. and U.K., and are more susceptible to ILD and RPILD. The mortality rate of patients with RPILD is extremely high. Conclusions The cases of JDM with positive anti-MDA5 antibody mainly presented with rash and mild muscle weakness, and could be complicated with ILD, pneumothorax and mediastinal emphysema without respiratory symptoms at early stage. Anti-MDA5 antibody titer is related to disease activity and can turn to negative after treatment.

Objective:To synthesize 177Lu-prostate-specific membrane antigen (PSMA)-617 with domestic 177Lu (made in China), and explore its optimal labeling condition, biodistribution, stability, and safety. Methods:177Lu-PSMA-617 was prepared with domestic 177Lu by a manual method. The optimal labeling condition, radiochemical purity, stability ( in vivo and in vitro), lipid-water partition coefficient, and plasma protein binding rate were determined. The uptake rate of 177Lu-PSMA-617 was evaluated by using 22RV1 cells. Biodistribution and SPECT/CT imaging were performed on normal mice with imported 177Lu-PSMA-617 as control group. The blood routine test was performed to evaluate the safety. Results:The best labeling result of domestic 177Lu-PSMA-617 can be obtained under the following conditions: pH=4.5, 100 ℃ for 30 min. And the radiochemical purity was ≥99%. The product was stable in vivo and in vitro, with the radiochemical purity >95% in 72 h. The plasma protein binding rate was (35.3±5.3)%, the lipid-water partition coefficient was -2.27±0.06, and the specific uptake rate of domestic 177Lu-PSMA-617 by 22RV1 cells reached the highest in 1 h ((7.58±0.84)%), which was slightly lower than the imported 177Lu-PSMA-617 ((7.86±0.96)%), but there was no significant difference between them ( t=-0.439, P>0.05). The distribution and SPECT/CT imaging of normal mice showed that domestic and imported 177Lu-PSMA-617 in blood were cleared quite fast, and both of them were excreted mainly through the kidneys. No obvious adverse reactions were found in the toxicity test of domestic and imported 177Lu-PSMA-617. There was no obvious abnormality in blood routine and liver and kidney metabolism. Conclusion:The domestic 177Lu-PSMA-617 has many advantages, such as qualified quality control, good biological properties and safety, which support its potential application value in diagnosis of prostatic neoplasms.

Objective To assess the effectiveness of the Mulliken's method in bilateral complete cleft lip patients with 3dMD system and anthropometric landmarks.Methods Thirty-one infants with bilateral complete cleft lip received treatment in Hospital of Stomatology,Wuhan University between January 2014 and December 2016.Patients underwent primary cheiloplasty and nasoplasty by the same senior surgeon.Periodic review was taken to measure and record the 7 items in the nasolabial area with three-dimensional (3D) images.Results The labial and nasal deformities were restored after primary surgery.The upper lip and nose were corrected and obtained the symmetric shape.The columella was elongated to acquire favorable nasal tip.The arc of nasal fornix was upward.The ridge of the white lipwas continuous and integrated with full vermilion tubercle.Total length of upper lip (Sn-Sto),thickness of the vermilion tubercle (Ls-Sto),protrusion of the nasal tip (Nh) and columellar height (Ch) were markedly improved and there were no significant difference between the observation group and the control group.The significant differences between two groups occurred in values of the nostril width (Nw),which was greater than control group and white lip height (Sn-Ls) and lower than that of control group.Conclusions Mulliken's method during the primary cheiloplasty of the bilateral complete cleft lip shows better results in correction the nasal deformity and the ideal effects are achieved during follow-up.