云克联合免疫抑制剂治疗活动期Graves''眼病患者

采用随机对照的方法，研究云克治疗方案，免疫抑制治疗方案以及联合治疗方案（云克联合免疫抑制剂）对活动期的GO患者的治疗效果。经过4个月治疗后，免疫抑制治疗方案（n=22)的总有效率为19／22，严重副作用的发生率为8／22；云克治疗方案（n=20)的总有效率17／20，严重副作用的发生率为2／20；联合治疗方案（n=24)的总有效率22／24，严重副作用的发生率为2／24；结果表明联合应用免疫抑制剂及云克治疗活动期的GO患者，既能取得较满意的疗效，又克服了长期应用激素治疗带来的副作用以及停用激素后的反弹。     

Metformin reduces morphine tolerance by inhibiting microglial-mediated neuroinflammation

Background

Tolerance seriously impedes the application of morphine in clinical medicine. Thus, it is necessary to investigate the exact mechanisms and efficient treatment. Microglial activation and neuroinflammation in the spinal cord are thought to play pivotal roles on the genesis and maintaining of morphine tolerance. Activation of adenosine monophosphate-activated kinase (AMPK) has been associated with the inhibition of inflammatory nociception. Metformin, a biguanide class of antidiabetic drugs and activator of AMPK, has a potential anti-inflammatory effect. The present study evaluated the effects and potential mechanisms of metformin in inhibiting microglial activation and alleviating the antinociceptive tolerance of morphine.

Methods

The microglial cell line BV-2 cells and mouse brain-derived endothelial cell line bEnd3 cells were used. Cytokine expression was measured using quantitative polymerase chain reaction. Cell signaling was assayed by western blot and immunohistochemistry. The antinociception and morphine tolerance were assessed in CD-1 mice using tail-flick tests.

Results

We found that morphine-activated BV-2 cells, including the upregulation of p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation, pro-inflammatory cytokines, and Toll-like receptor-4 (TLR-4) mRNA expression, which was inhibited by metformin. Metformin suppressed morphine-induced BV-2 cells activation through increasing AMPK phosphorylation, which was reversed by the AMPK inhibitor compound C. Additionally, in BV-2 cells, morphine did not affect the cell viability and the mRNA expression of anti-inflammatory cytokines. In bEnd3 cells, morphine did not affect the mRNA expression of interleukin-1β (IL-1β), but increased IL-6 and tumor necrosis factor-α (TNF-α) mRNA expression; the effect was inhibited by metformin. Morphine also did not affect the mRNA expression of TLR-4 and chemokine ligand 2 (CCL2). Furthermore, systemic administration of metformin significantly blocked morphine-induced microglial activation in the spinal cord and then attenuated the development of chronic morphine tolerance in mice.

Conclusions

Metformin significantly attenuated morphine antinociceptive tolerance by suppressing morphine-induced microglial activation through increasing AMPK phosphorylation.

     

HLA‐A,HLA‐B,HLA‐DRB1 allele and haplotype frequencies of 14 529 Chinese Han bone marrow donors living in Dalian,China

We investigated the allele and haplotype frequencies of HLA‐A, HLA‐B and HLA‐DRB1 loci in Dalian Chinese Han population using blood samples of unrelated marrow donors who live in Dalian. The genetic relationship between Dalian and different regions worldwide was further explored based on HLA status of different populations. A total of 14 529 samples were genotyped at 2‐digit level only by sequence‐specific oligonucleotide and sequence‐based typing methods. Allele frequencies of HLA‐A, HLA‐B and HLA‐DRB1 were calculated by the direct counting method. Haplotype frequencies and linkage disequilibrium (LD) values were calculated by the maximum likelihood method. F_ST values were calculated by allele frequency data of each locus. Phylogeny tree of Nei's D_A genetic distances was constructed by the UPGMA method. HLA‐A*02 was the most frequent allele at HLA‐A locus followed by A*11 and A*24. Alleles at HLA‐B locus ranked in decreasing order by frequency were B*40, B*15 and B*13. The three highest frequency alleles were DRB1*15, DRB1*09 and DRB1*12 at HLA‐DRB1 locus. A*30‐B*13‐DRB1*07 was the most frequent three‐locus haplotype. For the population relationships, Dalian had a relative close genetic relationship with Liaoning and Yantai‐Weihai and a relative distant genetic relationship with Australia. The information obtained in this study may provide useful information for anthropological studies, for disease‐association studies and helping bone marrow transplantation patients to search HLA‐matched donors.     

Prognostic Evaluation of Nasopharyngeal Carcinoma with Bone-Only Metastasis after Therapy

PurposeTo evaluate the prognosis of nasopharyngeal carcinoma (NPC) patients who developed bone-only metastasis after primary treatment and the stratification of these patients into different risk groups based on independent prognostic factors.ResultsThe median follow-up time was 15.5 months (range, 2–67 months) for the whole cohort. The median overall metastatic survival (OMS) time and the 2-year estimate OMS rate were 26.5 months and 52%, respectively. Multivariate analysis indicated that patients with short metastases-free interval, multiple bone metastases sites, high serum lactic dehydrogenase levels, and treated with radiotherapy or chemotherapy alone had significantly worse outcomes. Patients were stratified into three different risk groups based on the number of adverse factors present. The OMS curves of the three groups were all significantly different (p<0.001).ConclusionSeverl prognostic factors were found to be associated with worse outcomes. According to the number of adverse factors present, bone-only metastasis patients can be stratified into three risk groups with significantly different prognoses. Such grouping may help in improving the design of clinical trials and in guiding individualized treatment for NPC patients with bone-only metastasis.     

Nitric oxide inhibits spinally projecting paraventricular neurons through potentiation of presynaptic GABA release

Nitric oxide (NO) in the paraventricular nucleus (PVN) is involved in the regulation of the excitability of PVN neurons. However, the effect of NO on the inhibitory GABAergic and excitatory glutamatergic inputs to spinally projecting PVN neurons has not been studied specifically. In the present study, we determined the role of the inhibitory GABAergic and excitatory glutamatergic inputs in the inhibitory action of NO on spinally projecting PVN neurons. Spinally projecting PVN neurons were retrogradely labeled by a fluorescent dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocasbocyane (DiI), injected into the spinal cord of rats. Whole cell voltage- and current-clamp recordings were performed on DiI-labeled PVN neurons in the hypothalamic slice. The spontaneous miniature inhibitory postsynaptic currents (mIPSCs) recorded in DiI-labeled neurons were abolished by 20 microM bicuculline, whereas the miniature excitatory postsynaptic currents (mEPSCs) were eliminated by 20 microM 6-cyano-7-nitroquinoxaline-2,3-dione. Bath application of an NO donor, 100 microM S-nitroso-N-acetyl-penicillamine (SNAP), or the NO precursor, 100 microM L-arginine, both significantly increased the frequency of mIPSCs of DiI-labeled PVN neurons, without altering the amplitude and the decay time constant of mIPSCs. The effect of SNAP and L-arginine on the frequency of mIPSCs was eliminated by an NO scavenger, 2-(4-carboxypheny)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, and an NO synthase inhibitor, 1-(2-trifluoromethylphenyl) imidazole, respectively. Neither SNAP nor L-arginine significantly altered the frequency and the amplitude of mEPSCs. Under current-clamp conditions, 100 microM SNAP or 100 microM L-arginine significantly decreased the discharge rate of the DiI-labeled PVN neurons, without significantly affecting the resting membrane potential. On the other hand, 20 microM bicuculline significantly increased the impulse activity of PVN neurons. In the presence of bicuculline, SNAP or L-arginine both failed to inhibit the firing activity of PVN neurons. This electrophysiological study provides substantial new evidence that NO suppresses the activity of spinally projecting PVN neurons through potentiation of the GABAergic synaptic input.     

DNA polymerase mu gene expression in B-cell non-Hodgkin's lymphomas: an analysis utilizing in situ hybridization

DNA polymerase mu (pol mu) is a novel error-prone DNA repair enzyme bearing significant structural homology with terminal deoxynucleotidyltransferase. Whereas other human error-prone DNA polymerases identified thus far show no preferential lymphoid tissue distribution, the highest levels of pol mu mRNA have been detected in peripheral lymphoid tissues, particularly germinal center B cells. Conceivably, up-regulation of the pol mu gene may be biologically significant in lymphomagenesis, especially in the development of B-cell non-Hodgkin's lymphomas (B-NHLs), because of enhanced error-prone DNA repair activities. To explore this possibility, we generated a digoxigenin-labeled riboprobe to pol mu mRNA and used the probe and in situ hybridization to examine the expression pattern of the pol mu gene in formalin-fixed, paraffin-embedded tissue sections of 37 B-NHLs. This included eight chronic lymphocytic leukemia/small lymphocytic lymphomas, six mantle cell lymphomas, seven follicular lymphomas, nine diffuse large B-cell lymphomas, three splenic marginal zone lymphomas, two Burkitt's lymphomas, and two precursor B-lymphoblastic lymphomas. We also correlated the pol mu mRNA expression levels with the tumor proliferation index, which was assessed in each case by image analysis of Ki-67 immunostained slides. Nineteen of 21 (90%) B-NHLs arising from postgerminal center B cells (follicular lymphomas, diffuse large B-cell lymphomas, splenic marginal zone lymphomas, and Burkitt's lymphomas) exhibited high expression of pol mu mRNA. In contrast, only 2 of 16 (13%) B-NHLs arising from pregerminal center B cells (chronic lymphocytic leukemia/small lymphocytic lymphomas, mantle cell lymphomas, and precursor B-lymphoblastic lymphomas) expressed significant levels of pol mu mRNA. Pol mu gene expression did not seem to correlate with the proliferation index, especially because a significant level of pol mu mRNA was not detected in either case of precursor B-lymphoblastic lymphomas. In conclusion, pol mu gene expression is highly associated with B-NHLs of postgerminal center B-cell derivation. Furthermore, the expression level is independent of the proliferation rate and thus is unrelated to the biological aggressiveness of the tumors. These findings, along with the error-prone nature of the enzyme, suggest that up-regulation of pol mu gene expression may be a contributing factor to the pathogenesis of a subset of B-NHLs through DNA repair-associated genomic instability.     

自固化磷酸钙人工骨复合骨形成蛋白在牙槽骨缺损修复中的应用

目的探讨自固化磷酸钙人工骨(autosolidification calcium phosphate cement,ACPC)复合骨形成蛋白(bone morphogenetic protein,BMP)对牙槽骨缺损修复的生物学作用.方法将ACPC/BMP即刻植入拔牙术后患者牙槽创口,随访24周,通过临床检查和CCD数字化摄片观察牙槽骨缺损修复情况.结果实验位点无炎症、过敏和毒性反应发生,实验组患者牙槽嵴骨量吸收较空白对照组少,外形维持较好.结论BMP/ACPC复合骨兼具骨的引导性和诱导性,可促进新骨沉积钙化,即刻植入拔牙创利于增加牙槽骨量,维持牙槽外形,但材料降解性存在不足.     

HLA‐DPB1 allelic frequency of the Pumi ethnic group in south‐west China and evolutionary relationship of Pumi with other populations

A sequencing‐based typing of the HLA‐DPB1 gene was carried out in 51 unrelated healthy individuals from the Yunnan Pumi ethnic minority. A total of 18 DPB1 alleles, in which DPB1*0501 (52.0%) and DPB1*0402 (15.7%) greatly predominated, were found, of which alleles DPB1*20011, 2201, 3601, 3701, 3801, 4901, 5001 and 8001 were recorded for the first time in the Chinese population. This may be because the typing methods used in previous genotyping of Chinese populations were of lower sensitivity than that used in our study. A dendrogram constructed by the maximum likelihood method showed that the Pumi ethnic minority belongs to the Asian/Australasian cluster and has the closest relationship to Trobriander, implying an unusual relationship between Australasian and South China populations. However, the Yi ethnic minority, which also comes from the ancient Qiang, did not show a very close relationship with the Pumi. This is probably because the Pumi were historically assimilated by local south‐west China populations.     

五种玻璃化冻存试剂单独或组合应用对髓核细胞的毒性分析

BACKGROUND: Transplantation of allogeneic intervertebral disc can be facilitated by the cryopreservation of the intervertebral disc. But the traditional cryopreservation methods always lead to the appearing of ice crystals inside and outside the cells which can cause cellular injury. The vitrification method that can avoid the formation of ice crystals have been widely applied in the cryopreservation field. However, only a few reports have assessed the vitrified cryopreservation of the intervertebral disc, and the toxicity of cryoprotectants to the nucleus pulposus cells have not been fully explored. OBJECTIVE: To determine the order of toxicity of five commonly used cryoprotectants that are used alone or in combination to rabbit nucleus pulposus cells, and to select the optimal cryoprotectant for the vitrification of the intervertebral disc. METHODS: We chose five most commonly used cryoprotectants including dimethyl sulphoxide, formamide, ethylene glycol, propylene glycol and glycerol. Then, 5 single commonly used cryoprotectants, 10 mixed agents containing any 2 commonly used cryoprotectants, and 10 mixed agents containing any 3 commonly used cryoprotectants were formulated. Cell viability of nucleus pulposus cells was determined using cell counting kit-8 and fluorescein diacetate/propidium iodide method. All data obtained were analyzed statistically to choose the appropriate combining scheme with less toxicity. RESULTS AND CONCLUSION: The order of the toxicity of these five commonly used cryoprotectants from low to high was ethylene glycol, glycerol, formamide, dimethyl sulphoxide, and propylene glycol. The toxicity of the combined agents containing two or three commonly used cryoprotectants was lower than that of any commonly used cryoprotectants that were used to formulate them. The toxicity of the mixed agents that contained ethylene glycol or glycerol was lower than that of any other mixed agents. So we can choose the mixed cryoprotectants that contain ethylene glycol and (or) glycerol for the vitrification of the intervertebral disc.       

脊髓康对脊髓损伤大鼠Nogo-66受体NgR表达的影响

文题释义：   文题释义：脊髓损伤：是指由于外界直接或间接因素导致脊髓损伤,在损害的相应节段出现各种运动、感觉和括约肌功能障碍,肌张力异常及病理反射等的相应改变。脊髓损伤的程度和临床表现取决于原发性损伤的部位和性质。脊髓损伤可分为原发性脊髓损伤与继发性脊髓损伤。前者是指外力直接或间接作用于脊髓所造成的损伤。后者是指外力所造成的脊髓水肿、椎管内小血管出血形成血肿、压缩性骨折以及破碎的椎间盘组织等形成脊髓压迫所造成的脊髓的进一步损害。Nogo受体NgR：是Nogo-A发挥作用的受体,是一种糖基磷脂酰肌醇锚定膜蛋白,能与细胞外结构域Nogo-66结合,并被激活,激活的NgR启动神经细胞内的信号转导通路抑制轴突再生和结构的重塑性。  背景：研究发现,脊髓损伤后髓鞘相关抑制分子的产生是影响轴突再生微环境的重要原因。中医药具有多因素、多靶点的优点,正逐步成为改善神经再生微环境的研究热点。目的：探索中药脊髓康对脊髓损伤后Nogo-66受体NgR表达的影响。方法：144只大鼠随机等分为6组,假手术组、模型组、强的松组、脊髓康高、中、低剂量组,每组24只。后5组以改良Allen’s法建立脊髓损伤动物模型。强的松组大鼠每天给予0.06 g/kg醋酸泼尼松灌胃,1次/d。脊髓康低中高剂量组大鼠每天给予12.5,25,50 g/kg脊髓康灌胃,1次/d。假手术组和模型组大鼠每天给予20 mL生理盐水灌胃,1次/d。各组动物均连续给药至动物处死。结果与结论：与模型组相比,强的松组及脊髓康低中剂量组大鼠脊髓组织中NgR蛋白及mRNA表达水平均显著降低。提示脊髓康可促进大鼠脊髓损伤后神经功能的恢复,可有效抑制脊髓损伤区NgR蛋白的表达,从而抑制不利于神经再生的因素,进一步改善神经再生的微环境。中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程 ORCID:0000-0002-8518-6160(郭杨)