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【Objective】 To construct an acute toxoplasma encephalitis mouse model by observing the pathological changes in the hippocampus of mice infected with Toxoplasma gondii strain RH. 【Methods】 The quantitative RH Toxoplasma gondii (100, 500, and 1 000 trophozoites) were injected into the hippocampal CA1 region of mice by the stereotaxic surgery; the survival status of mice was observed. Giemsa staining was used to observe the changes of toxoplasma in mouse ascites and brain tissue homogenates. Nissl staining and HE staining were used to observe the pathological changes of hippocampal nerve tissue. The distribution of Toxoplasma gondii in brain tissue was observed by immunohistochemical ABC method. 【Results】 The RH Toxoplasma gondii infected mice showed obvious symptoms such as arched back, bristling hair, abdominal distension, subtle tremor and hemiplegia on the fourth day of infection. The survival of mice in 100 trophozoites group was longer, no trophozoites of Toxoplasma gondii were found in ascites, a few pseudocysts were found in brain tissue homogenates after infected for 96 hours, and more trophozoites were found after death. Nysl staining and HE staining showed more tissue necrosis foci and loss of nerve cells in CA1 area after infected 144 h. The injury aggravated with the prolongation of infection time. Toxoplasma trophozoites were found in ascites and brain homogenates of mice in 500 and 1000 trophozoites groups. Nissl staining revealed neuronal loss and massive necrosis in the hippocampus. HE staining showed necrosis and inflammatory cell infiltration. The brain tissue injury significantly aggravated compared with 100 trophozoites group. The distribution of Toxoplasma gondii in the necrotic foci was confirmed by immunohistochemistry. 【Conclusion】 The survival of 100 trophozoite mice infected with Toxoplasma gondii strain RH was longer, and the pathological changes of brain tissue gradually aggravated. The damage was relatively confined to the brain tissue, and the mice showed typical symptoms of toxoplasma encephalitis. Therefore, the mouse model of acute toxoplasma encephalitis can be constructed by localized infection of 100 toxoplasma trophozoites, which can lay a foundation for future research on the mechanism of toxoplasma injury to cranial nerves.
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Adenocarcinoma of esophagogastric junction (AEG) has attracted more attention in recent years. Surgical method of Siewert type Ⅱ AEG is especially controversial, mainly focusing on the scope of lymph node dissection, safety of surgical margin, and digestive tract reconstruction. The abdominal transhiatal approach and right thoracoabdominal Ivor-Lewis approach are the main surgical approaches of totally laparoscopic or thoracoscopic surgery for Siewert type Ⅱ AEG, which not only need close teamwork, but also require rich experience in laparoscopic surgery. The authors has started to choose these two totally minimally invasive surgical approaches, the feasibility and safety of which are proved. The key surgical details are presented in this article for reference.
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Chemotherapy is among the limited choices approved for the treatment of hepatocellular carcinoma (HCC) at intermediate and advanced stages. Preferential and prolonged drug exposure in diseased sites is required to maximize the therapeutic index of the drug. Here, we report an injectable supramolecular peptide hydrogel as an intraperitoneal depot for localized and sustained release of triptolide for the treatment of orthotopic HCC. We chose peptide amphiphile C-GNNQQNYKD-OH-based nanofibers as gelators and carriers for triptolide. Sustained triptolide release from the hydrogel was achieved over 14 days , with higher accumulation in and cytotoxicity against human HCC Bel-7402 in comparison with L-02 fetal hepatocytes. After intraperitoneal injection, the hydrogel showed prolonged retention over 13 days and preferential accumulation in the liver, realizing HCC growth inhibition by 99.7 ± 0.1% and animal median survival extension from 19 to 43 days, without causing noticeable pathological changes in the major organs. These results demonstrate that injectable peptide hydrogel can be a potential carrier for localized chemotherapy of HCC.
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Objective To study the the feasibility of intestinal derotation maneuver in the resection of gastrointestinal neoplam localized in the vicinity of the ligament of Treitz.Method Intestinal derotation maneuver was applied in all the 9 case of gastrointestinal neoplasms near the ligament of Treitz,from January 2014 to January 2016,at the Second Hospital of Jilin University.The clinical date were retrospectively analyzed.Results Intestinal derotation maneuver were applied without failure in all the nine patients,the mean operation time were (195 ± 50) min,and the mean intestinal derotation maneuver time were (19 ±5) min;No derotation procedure-related injury occured,and the mean estimated blood loss were (132 ±94) ml.Early postoperative ileus developed in one case and gastroparesis in two cases.All were recovered by conservative treatment.The follow-up period were 3-24 month,tumor recurrence occurred in two cases and one case died.One case lossed to follow-up after postoperative 3 month.Conclusion The present data suggest that clinical use of intestinal derotation maneuver was feasible for surgical management of gastrointestinal neoplasms near the ligament of Treitz.
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Bacterial two-hybrid system is a newly developed method for studying protein-protein interactions. However, in our studies of the interaction of regulatory proteins in Streptomyces, it was found that the bacterial two-hybrid system is not sensitive enough by the blue-and-white selection on X-gal plate. To overcome this drawback, the reason of false positive clone was firstly determined, which was the disturbance of other direct or indirect regulation on lacZ promoter. Then the disturbance was diluted by introducing multicopy lacZ promoter, which drive another reporter gene gfp. By such design, the sensitivity of the modified bacterial two-hybrid system was significantly inproved and the two different reporters also help to decrease the rate of the false positive clones. Further the evaluation of the modifiedd bacterial two-hybrid system indicated that the sensitivity was significantly improved.
Subject(s)
Bacteria , Genes, Reporter , Promoter Regions, Genetic , Protein Interaction Mapping , Methods , Two-Hybrid System TechniquesABSTRACT
Gene expression exhibits temporal and spatial patterns to response environmental changes and growth cycle. Gene expression is under strict control at different levels among which control at transcription level is the predominant mode, especially in prokaryotes. In this review, we summarized the new developments of methods used in transcriptional studies, including modifications and improvements of the classic methods, such as gel-shift assay, DNA foot printing, and in vivo reporter system. In addition, we introduced examples to apply new methods, such as surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) to characterize protein-DNA, ligand-protein, and ligand-protein-DNA interactions. The collection of these methods and their application could guide and accelerate relevant studies.
Subject(s)
Calorimetry , DNA Footprinting , Gene Expression , Ligands , Proteins , Surface Plasmon Resonance , Transcription, GeneticABSTRACT
Objective To examine whether microinjectlon of morphine into the rat thaiamle nucleus submedlus (Sin) could depress the bee venom (BV)-induced nociceptive behaviours. Methods In inflammatory pain model induced by BV subcutaneous injection into rat unilateral hind paw, the inhibitory effects of morphine microinjection into thalamic nucleus suhmedius (Sin) on the spontaneous nociecptlve behavior, heat hyperalgesia and tactile ailodynia, and the influence of naioxone on the morphine effects were observed in the rat. Results A single dose of morphine (5.0 μg, 0. 5μL) applied into the Sm ipsilaterni to the BV injected paw significantly depressed the spontaneous paw flinching response. Morphine also significantly increased the heat paw withdrawal iateneies in the bilateral hind paw and the tactile paw withdrawal threshold in the ipsilnteral hind paw 2 hours after BV injection. All these depressive effects could be effectively antagonized by pre-treatment with the opiuld receptor antagonist naloxone (1.0μg, 0. 5μL) in the Sm 5rain prior to morphine administration. Naloxone alone injected to the Sm had no effect on the BV-induecd nociceptive behavior. Conclusion These results suggest that Sm is involved in opioid receptor-mediated antt-nociception in the rat with the BV-induced inflammatory pain. Together with results from previous studies, it is likely that this effect is produced by activation of the Sm-ventrolateral orbital cortex-periaqueductal gray pathway, leading to activation of the brainstem descending inhibitory system and depression of the nodceptive inputs at the spinal cord level.
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Objective To find out the possible regularity and mechanism of the adaptable change of human being T lymphocytes for physical exercise with oxygen and bring the original data for the Movement of All People Improving their Health. Methods We selected 16 untrained female students in university and let them had the same amount of exercise for 8 weeks. After that, we collected the cycle blood at the time point of before exercise, the end of exercise and 1 hour after exercise at the end of the 0,first,2 nd,4 nd,6 nd and 8 nd week respectively, so as to determine its stimuli index (SI) by MTT method. Results In the different time sect, such as the early stage of exercise, quiet condition,as soon as the end of exercise and 1 hour after exercise, we found that the SI were obviously Iower than that of normal (P＜0. 05) ,especially in the time sect of the end of exercise. Continuing to 4 weeks,the function of T lymphocytes restored gradualy and it lasted to the 8 th week, the SI in quiet condition and 1 hour after exercise had restored to normal(P＞0.05),but in the end of exercise, it still was Iow,however, the extent of the cases selected was in a condition of acute excitability. Conclusion As the bodies adapting to the exercise, the function of T lymphocytes restored slowly and the rate increased faster and faster.
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Objective To establish a speed and effective method to detect rotavirus. Methods Using ELISA and one step RT-PCR to detect 196 clinic samples from Xi'an area. Results Compared with ELISA method, one step RT PCR was more sensitive and specific (P ＜0.05). Conclusion One step RT-PCR is a simple, speed, sensitive and spe cific method for clinic and epidemic studies of rotavirus.
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Objective To investigate the analgesic effect and possible mechanisms of interleukin-2(IL-2) on the neuropathic pain,and to screen the novel drug for neuropathic pain diseases.Methods Male C57BL/6 mice whose unilateral L5/L6 spinal nerves were ligated were used as a neuropathic pain model.The pain thresholds of paw ipsilateral to the nerve injury before and after intraperitoneal(i.p.) injection of different doses of IL-2(1.0?10~(6)u/kg,2.5?10~(6)u/kg and 5.0?10~(6)u/kg) were measured by the mechanical and cold stimuli.Opioid receptor antagonist naloxone(1mg/kg),30min prior to the administration of 5.0?10~(6)u/kg of IL-2,was given to investigate the influence of naloxone on the analgesic effect induced by IL-2.Results IL-2 produced dose-dependent analgesic effect.Both 5.0?10~(6)u/kg and 2.5?10~(6)u/kg of IL-2 significantly increased the 50% paw withdrawal threshold((P
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Objective To explore the influence of chronic neuropathic pain on cellular immune function of mice models.Methods The mice models were established by ligating tibeal nerve and common peroneal nerve on one side,and the influence of chronic neuropathic pain on cellular immune function and tumor necrosis factor-?(TNF-?) and interleukin-6(IL-6) was observed with lymphocyte cell increase experiment(MTT method) and enzyme linked immunosorbent assay(ELISA).Results The increased reactivity of T lymphocyte of the model group was significantly higher than that of the sham group and the control group.At the same time,content of TNF-? and IL-6 in blood of the model group also increased.Conclusion Cellular immune function of mice is increased in the state of chronic neuropathic pain.