Purpose To construct expression vector of Fas extracellular region gene(eFas) ,to express and purify recombination protein and to prepare polyclonal antibody, which have laid a foundation of studying its function. Methods The eFas gene encoding sequence was acquired through overlapping PCR, and pET-22b ( + )/eFas expression vector was constructed. Then this vector was transformed into E. coli Rosetta-gami. Re-combinant protein was expression being induced by IPTG,and was purified using Ni-NTA matrix of affinity chromatograph. The purity of recombination protein was identified by SDS-PAGE. Hereafter, the purified eFas recombinant protein was immunized to New Zealand white rabbit in order to prepare polyclonal antibody. The titer of polyclonal antibody was determined by ELISA. Results The encoding sequence and expression vector of eFas was obtained while the interest protein was mainly expressed in the inclusion body. The eFas fusion protein's expression quantity accounts for more than 30% proportion of total E. coli protein. The eFas protein we obtained was provided with the purity of at least 95 % . Conclusion The successful constrution, expression and purification of FasL fusion protein and preparation of polyclonal antibody will provide some material for further studies of Fas.

Objective:The purpose of this study is to evaluate the effects of a single-chain antibody against death receptor 5 (ZF1) on tumor growth and survival in murine H22 hepatocellular carcinoma tumor model.Methods:Killing effect of ZF1 on H22 cells was analyzed by MTT assay in vitro. The apoptosis rate of H22 cells induced by ZF1 was detected using Flow Cytometry assay. The transplanted model of H22 tumor was developed in mice. The mice were randomly divided into four groups, PBS group, ZF1 group, EPI group and combined treatment group of ZF1/EPI. Tumor growth and body weight changes were observed. After treatment over 13 days, the tumor tissue for HE staining and TUNNEL assay was performed to detect apoptosis.Results:The results showed that ZF1 could inhibit growth of H22 cells in a dose dependent manner. The growth inhibition rate was up to 84.5%. The results showed that ZF1 alone or in combination with ZF1/EPI, the tumor growth was significantly inhibited. HE staining and TUNNEL analysis showed that ZF1 could effectively induce apoptosis of tumor cells without toxic effects, especially in ZF1/EPI combined treatment group.Conclusion:It is showed that ZF1 induces a good inhibition on the proliferation of H22 cell, especially in combined treatment group of ZF1/EPI.

Objective To explore the effect of TGF-β microbubble targeting on the treatment of skeletal muscle injury in rats.MethodsEighty SD rats of 2 months old were randomly divided into experimental group(including TGF-β microbubble),pure drug group,microbubble group(exclusive drug) and control group,20 SD rats in each group.Four groups accepted ultrasonic transmission with consistent time and frequency.The contraction stress and stress relaxation of injured gastrocnemius muscle were measured,the changes of injured gastrocnemius muscle were observed by light microscope and electron microscope.Results There was no statistically significant difference in contraction stress of gastrocnemius muscle among all groups at 1,5,9 days after injury(P>0.05).And at 14 days,the contraction stress of gastrocnemius muscle of the experimental group and the pure drug group were obviously higher than those of the rest two groups,the differences were significant(P<0.05).The difference of gastrocnemius muscle contraction stress was statistically significant between the experimental group and the pure drug group(P<0.05).For the stress relaxation of gastrocnemius muscle,the experimental group was lower than the other three groups at 1 day after injury,the differences were significant(P<0.05).Light microscopy showed the number of muscle fiber in the experimental group increased obviously at 9 days after injury.There was no scarring at 14 days after injury.Electron microscope showed mitochondria and sarcoplasmic reticulum increased in the experimental group,and the new muscle cells and satellite cells were significantly more than those of the rest three groups.Conclusion TGF-β cell targeted therapy can obviously improve the shrinkage stress of skeletal muscle in rats after injury and recover the stress decay,which can obviously promote the injury repair and regeneration of skeletal muscle,and reduce scar formation at the same time.

Objective To evaluate the Flavivirus specific monoclonal antibody(McAb) 2A10 as detective antibody for simultaneously identify tick borne encephalitis virus( TBEV), Japanese encephalitis virus( JEV), dengue ( DEN )-2, DEN-4 and yellow fever virus ( YFV ) by antibody microarray technique.Methods The antibody microarray was developed by spotting TBEV, JEV, DEN-2, DEN-4 and YFV specific McAb on chip as capture antibodies. After incubating with cultured viral supernatants of the above viruses, CY3 labeled detective antibody 2A10 was added to the chips. After reaction, the antibody microarray was scanned and the results were analyzed. By comparing the signal intensities of different spots on chips,the detecting titre and sensitivity of 2A10 for Flavivirus were determined, and the value of 2A10 in detection of Flavivirus was evaluated. Results The hybridization results demonstrated that the titre of 2A10 for Flavi2A10 was specific for Flavivirus and could be used as universal detective antibody for Flavivirus on antibody microarray.

ObjectiveTo evaluate the efficacy and safety of rifaximin in the prevention of spontaneous bacterial peritonitis (SBP). MethodsCNKI, Wanfang Data, CBM, PubMed, Embase, and Cochrane Library were searched for randomized controlled trials (RCTs) and cohort studies on rifaximin in the prevention of SBP published up to July 5, 2020. The articles were screened according to the inclusion and exclusion criteria, and data extraction and quality assessment were performed. RevMan 5.3 software was used to conduct the meta-analysis. Results A total of 13 studies (with 2207 patients in total) were included, among which there were 6 RCTs and 7 cohort studies. The results of the meta-analysis showed that compared with the non-prevention group, the rifaximin group had significantly lower incidence rate of SBP (odds ratio ［OR］=0.36, 95% confidence interval ［CI］: 0.14-0.96, P=0.04) and mortality rate (OR=0.59, 95% CI: 037-0.95, P=0.03); compared with the norfloxacin group, the rifaximin group had significantly lower incidence rate of SBP (OR=039, 95% CI: 025-0.62, P＜0.001), mortality rate (OR=0.55, 95% CI: 0.34-0.92, P=0.02), and adverse reactions (OR=0.36, 95% CI: 0.22-059, P＜0.001). The subgroup analysis based on the type of prevention showed that there was no significant difference in primary prevention between the two groups (OR=0.56, 95% CI: 0.23-1.35, P=0.20), and in secondary prevention, the rifaximin group had a significantly lower incidence rate of SBP (OR=0.18, 95% CI: 0.08-0.43, P＜0.001). In addition, it was also found that rifaximin significantly reduced the incidence rate of hepatorenal syndrome (OR=0.34, 95% CI: 0.15-0.77, P=0.01) and hepatic encephalopathy (OR=0.55, 95% CI: 0.32-0.95, P=0.03). ConclusionRifaximin is safe and effective for the primary and secondary prevention of SBP. Rifaximin is superior to norfloxacin in secondary prevention, which still needs to be confirmed by high-quality multicenter RCTs.

Objective:To investigate the clinical effectiveness of one-stage biplanar osteotomy correction of angulation and shortening deformity of the lower extremity after epiphyseal injury of the distal femur with a monorail external fixator.Methods:The data of 5 patients (2 males and 3 females) with angulation and shortening deformity in the lower extremity after epiphyseal injury of the distal femur were retrospectively analyzed.The patients underwent monorail external fixator assisted one-stage osteotomy correction of the distal femur and distraction osteogenesis of the middle and upper femur in Zhengzhou Orthopeadics Hospital from May 2017 to December 2019.The mean age was 13.6 years old (range: 10 to 17 years old). The affected limbs were shortened by 5.1 cm on average (range: 3.9 to 6.5 cm). The average angulation deformity of the distal femur was 24.9° (range: 17.0°to 30.5°). The mechanical lateral distal femoral angle (mLDFA), the mechanical posterior distal femoral angle (mPDFA), the mechanical axis deviation (MAD), the range of motion (ROM) of the knee, and the length of the lower limbs before surgery and at the final follow-up were measured and analyzed.Results:All patients were followed up for 22 months on average (range: 15 to 32 months). For all the 5 patients, the mechanical axis was well realigned, mLDFA, mPDFA, and MAD returned to normal range, and the length of the affected limb achieved the goal as planned before the surgery.Besides, the affected limbs were lengthened by 5.6 cm on average (range: 3.9 to 8.0 cm), and the median healing index was 35.6 d/cm (range: 29.0 to 45.0 d/cm). The bone callus in the distraction area and the osteotomy end were well healed at the final follow-up, as indicated by the X-ray results.At the end of the distraction period, the flexion ROM of the knee in all patients reached basically 90°.By the final follow-up, all patients had a normal knee ROM.No vascular or nerve injury, dislocation of hips or knee joints, re-fracture after disassembly, deep infection and other complications were observed in all patients.Conclusions:One-stage biplanar osteotomy correction of angulation and shortening deformity of the lower extremity after epiphyseal injury of the distal femur with a monorail external fixator is safe and feasible.The method requires no multiple operations and improves the tolerance of patients during the treatment period.

Objective To establish the chronic low flow myocardial hibernation animal model in pigs, and to assess the diagnostic value for myocardial hibernation by using various imaging methods. Methods A total of 13 miniswine (30-40 kg) were used. All animals underwent general anesthesia and orotracheal intubation while the animals were mechanically ventilated. Under sterile conditions, left ventriculography and coronary angiography were performed by introduction of catheter into the right femoral artery. Further, a left anterolateral thoracotomy was performed in the third intercostal space. The proximal LCX was dissected free to allow placement of an ameroid constrictor. More than 1 month later, left ventriculography and coronary angiography were performed again, followed by cine MRI at rest and during stress with low dose of dobutamine (5 ?g?kg -1 ?min -1 ), respectively. Traditional and/or breath hold cine MRI were used to evaluate regional left ventricular wall motion, corresponding to basal, midventricular and apical short axis tomograms. Regional wall motion score index (WMSI) was calculated. At the same time 99m Tc MIBI myocardial SPECT was performed at rest and during nitroglycerin administration, respectively. All animals were finally sacrificed for pathological examination. Triphenyl tetrazolium chloride (TTC) staining was used to assess the myocardial infarction. Electron microscopy was used to identify myocardial cellular changes characteristic of hibernating myocardium. Results Three pigs died during surgery or within two weeks after surgery. One pig died of anesthesia during SPECT examination, 1 pig suffered from aneurysm, and another one pig showed negative findings. The other 7 pigs were found with hypokinetic ( n =4) or akinetic ( n =3) myocardial regions related to stenosed LCX (70%-99%). Resting cine MRI demonstrated decreased regional motion of the lateral and posteroinferior walls (ischemic regions) of the left ventricle ( n =7), compared with the nonischemic anteroseptal regions; but the low dose dobutamine (5 ?g?kg -1 ?min -1 ) could recover those hypokinetic or akinetic myocardial regions, characteristic of hibernating myocardium. Resting 99m Tc MIBI myocardial SPECT ( n =6) showed a fixed perfusion defect on the corresponding ischemic areas, which became reversible on the nitrate augmented myocardial perfusion imaging. It also indicated myocardial viability presented at the ischemic areas. TTC staining revealed patchy infarction of the area at risk localized to the endocardial surface ( n =3), and no myocardial infarction ( n =4). Electron microscopy of sections from the hibernating regions revealed loss of contractile materials, increased numbers of small mitochondria, and glycogen accumulation within viable cardiomyocytes, which had been described as hallmarks of hibernating myocardium. Conclusion Chronic low flow myocardial hibernation can be reproduced in an animal model during progressive coronary stenosis caused by ameroid constrictor.

Cells can adapt to environment and development by reconstructing their transcriptional networks to regulate diverse cellular processes without altering the underlying DNA sequences. These alterations, namely epigenetic changes, occur during cell division, differentiation and cell death. Numerous evidences demonstrate that epigenetic changes are governed by various types of determinants, including DNA methylation patterns, histone posttranslational modification signatures, histone variants, chromatin remodeling, and recently discovered chromosome conformation characteristics and non-coding RNAs (ncRNAs). Here, we highlight recent efforts on how the two latter epigenetic factors participate in the sophisticated transcriptional process and describe emerging techniques which permit us to uncover and gain insights into the fascinating genomic regulation.
Cell Death ; Cell Differentiation ; Cell Division ; Chromatin ; chemistry ; metabolism ; Chromatin Assembly and Disassembly ; DNA Methylation ; Epigenesis, Genetic ; Eukaryotic Cells ; cytology ; metabolism ; Histones ; genetics ; metabolism ; Humans ; Protein Processing, Post-Translational ; RNA, Untranslated ; genetics ; metabolism ; Transcription, Genetic

Objective:To investigate the value of three-dimensional (3D) CT in diagnosing cricoarytenoid dislocation.Methods:From January 2021 to December 2021, 31 patients with unilateral cricoarytenoid dislocation who had been treated by reduction forceps at the Affiliated BenQ Hospital of Nanjing Medical University were collected retrospectively, and their voice recovered or improved significantly after therapy. The preoperative CT images were reconstructed by volume rendering (VR). The dislocated side (left and right), type of dislocation (total dislocation and subluxation), and dislocation direction (anterior, posterior, internal and external dislocation) of cricoarytenoid dislocation were observed. According to arytenoid articular surface of cricoid cartilage exposed completely or not (caused by arytenoid displacement), they were divided into complete dislocation and subluxation. According to the direction of arytenoid displacement and the part of arytenoid articular surface of cricoid cartilage exposed, they were divided into anterior, posterior, internal and external dislocation. According to the shape of the vocal cords on laryngoscope, anterior and posterior dislocation of each case was judged, and then compared with that of CT.Results:On VR images, there were 28 cases of cricoarytenoid subluxation (90.3%, 28/31) and 3 cases of complete dislocation (9.7%, 3/31). Left cricoarytenoid dislocation was 26 cases (83.9%, 26/31) and right cricoarytenoid dislocation was 5 cases (16.1%, 5/31). Posterior dislocation was 28 cases (90.3%, 28/31) and anterior dislocation was 3 cases (9.7%, 3/31). There were 23 cases of internal dislocation (74.2%, 23/31), 2 cases of external dislocation (6.4%, 2/31), and 6 cases without obvious internal and external dislocation (19.4%, 6/31). Three cases of complete dislocation were left posterior internal dislocation.There were 24 cases of left posterior dislocation (77.4%, 24/31), 4 cases of right posterior dislocation (12.9%, 4/31), 2 cases of left anterior dislocation (6.4%, 2/31) and 1 case of right anterior dislocation (3.2%, 1/31). On laryngoscope, there were 19 cases of posterior dislocation (61.3%, 19/31), 9 cases of anterior dislocation (29.0%, 9/31), 3 cases were difficult to assess (9.7%, 3/31) because of aryepiglottic fold covering. Sixteen cases (55.2%, 16/28) were consistent with 3D CT, and 12 cases (42.8%, 12/28) were inconsistent.Conclusion:The 3D CT is a reliable method to evaluate cricoarytenoid dislocation, which can show dislocated side, type and direction of cricoarytenoid dislocation clearly.

Objective To study the diagnostic value and clinical efficacy of endoscopic ultrasound-guided fine needle aspiration ( EUS-FNA ) for mediastinal and abdominal lymphadenopathy. Methods Thirty patients who underwent EUS-FNA for mediastinal or abdominal lymphadenopathy between May 2009 and December 2015 were reviewed. The clinical efficacy of EUS-FNA was evaluated by pathological results and the follow-up. The EUS-FNA effect on clinical decision was also analyzed. Results Lesions were located in the mediastinum in 10 cases and in the abdomen in 20 cases. The total diagnostic accuracy, sensitivity, specificity, positive predictive value ( PPV) and negative predictive value ( NPV) of EUS-FNA were 96. 7%, 94. 7%, 100. 0%, 100. 0% and 91. 7%, respectively. Of all the 30 cases, 20 lymph glands were of unknown origin. The diagnostic accuracy, sensitivity, specificity, PPV and NPV of EUS-FNA in these lesions were 95. 0%, 88. 9%, 100. 0%, 100. 0% and 91. 7%, respectively. The combination of cytological and histological examination had higher accuracy ( 96. 7% VS 73. 3%, P=0. 026) and sensitivity ( 94. 7%VS 57. 8%, P= 0. 019 ) than cytological examination only. Immunohistochemistry stains were performed in 12 neoplastic cases, and 11 ( 91. 7%) were confirmed. The diagnosis by EUS-FNA had positive impact on clinical decisions in 27 patients ( 90. 0%) . Conclusion EUS-FNA is an effective approach for mediastinal and abdominal lymphadenopathy, and the result has a positive impact on clinical decisions. The combination of cytological and histological examination and application of ancillary techniques, such as immunohistochemistry stains, can improve the diagnostic efficacy of EUS-FNA.