Nucleolar localization of non-structural protein 3b, a protein specifically encoded by the severe acute respiratory syndrome coronavirus

The open reading frame 3 (ORF3) of the severe acute respiratory syndrome coronavirus (SARS-CoV) genome encodes a predicted 154-amino acid protein, which lacks similarities to any known protein, and is named 3b. In this study, it was shown that 3b protein was predominately localized to nucleus with EGFP tag at its N- or C-terminus. The localization patterns were similar in different transfected cells. Immuno-fluorescence assay revealed that 3b protein was co-localized well with C23 in nucleolus. C23, B23 and fibrillarin all are important nucleolar proteins, which localize in the region of the nucleolus. Co-transfection of p3b-EGFP with pC23-DsRed, pB23-DsRed and pfibrillarin-DsRed further confirmed 3b's nucleolus localization. With construction of serial truncated mutants of 3b, a region (residues 134-154 aa) responsible for nucleolar localization was determinated in 3b protein. These results provide a new insight for further functional studies of SARS-CoV 3b protein.     

腺样体肥大患儿血清炎症介质和淋巴细胞亚群表达特征及与患儿预后的相关性

目的研究腺样体肥大患儿血清炎症介质、淋巴细胞亚群表达特征及其与患儿预后的相关性。方法选取浙江中医药大学附属第三医院收治的86例腺样体肥大患儿(观察组)和86例健康儿童(对照组)作为研究对象,采集血液标本测定血清炎症介质及淋巴细胞亚群相关指标,对比两组儿童的测定结果。分析腺样体肥大患儿的肥大程度、腺样体再增生情况,对比不同肥大程度患儿的血清指标,并分析各指标与患儿病情程度及预后(腺样体再增生)的关系。结果观察组患儿肿瘤坏死因子-α(TNF-α)、可溶性白细胞介素-2受体(sIL-2R)及白细胞介素-6(IL-6)水平分别为(492.44±50.56)mg/L、(420.48±61.79)U/ml及(33.65±4.51)pg/ml,对照组儿童分别为(301.22±41.65)mg/L、(236.55±57.90)U/ml及(16.25±3.27)pg/ml,差异均有统计学意义(t=16.004,P<0.05;t=20.311,P<0.05;t=8.995,P<0.05)。观察组CD4⁺、CD8⁺及CD4⁺/CD8⁺水平分别为(35.75±7.05)、(16.22±2.69)及(1.46±0.67),对照组分别为(34.25±4.53)、(17.62±2.35)及(1.40±0.62),差异均无统计学意义(t=1.036,P>0.05;t=0.905,P>0.05;t=0.823,P>0.05)。相比腺样体中度肥大患儿,重度肥大患儿的TNF-α、sIL-2R、IL-6、CD4⁺及CD4⁺/CD8⁺水平明显更高(P<0.05)。相比未增生患儿,腺样体再增生患儿的TNF-α、sIL-2R、IL-6、CD4⁺及CD4⁺/CD8⁺水平明显更高(P<0.05)。相关分析显示,患儿腺样体肥大程度与TNF-α、sIL-2R、IL-6、CD4⁺及CD4⁺/CD8⁺水平均呈正相关(P<0.05)。患儿腺体再增生与TNF-α、sIL-2R、IL-6、CD4⁺及CD4⁺/CD8⁺水平也呈正相关(P<0.05)。结论腺样体肥大患儿的血清炎症介质及部分T淋巴细胞亚群表达显著升高,其与腺样体肥大程度、患儿预后密切相关。     

Correction to: Cichorium intybus L. promotes intestinal uric acid excretion by modulating ABCG2 in experimental hyperuricemia

The study of sex differences in hyperuricemia can provide not only a theoretical basis for this clinical phenomenon but also new therapeutic targets for urate-lowering therapy. In the current study, we aimed to confirm that estradiol can promote intestinal ATP binding cassette subfamily G member 2 (ABCG2) expression to increase urate excretion through the PI3K/Akt pathway. The estradiol levels of hyperuricemia/gout patients and healthy controls were compared, and a hyperuricemia mouse model was used to observe the urate-lowering effect of estradiol and the changes in ABCG2 expression in the kidney and intestine. In vivo and in vitro intestinal urate transport models were established to verify the urate transport function regulated by estradiol. The molecular pathway by which estradiol regulates ABCG2 expression in intestinal cells was explored. The estradiol level of hyperuricemia/gout patients was significantly lower than that of healthy controls. Administering estradiol benzoate (EB) to both male hyperuricemic mice and female mice after removing the ovaries confirmed the urate-lowering effect of estradiol, and hyperuricemia and estradiol upregulated the expression of intestinal ABCG2. Estradiol has been confirmed to promote urate transport by upregulating ABCG2 expression in intestinal urate excretion models in vivo and in vitro. Estradiol regulates the expression of intestinal ABCG2 through the PI3K/Akt pathway. Our study revealed that estradiol regulates intestinal ABCG2 through the PI3K/Akt pathway to promote urate excretion, thereby reducing serum urate levels.     

口腔固定修复学临床前期实习中的问题与对策

本文总结了北京大学口腔医学院3届八年制口腔医学专业学生口腔固定修复学临床前期实习教学的经验,针对实习中出现的典型问题进行了分析,采取了相应的对策和方法,取得了良好的效果.     

基于电子鼻技术的不同品质鹿茸饮片气味特征分析

目的:基于电子鼻技术对不同品质鹿茸饮片气味特征进行分析与表征。方法:采用PEN3型电子鼻系统,分析22批鹿茸样品的气味特征,以对传感器响应值为指标,进行传感器区别贡献率分析(Loadings)、主成分分析(PCA)、线性判别分析(LDA)。结果:Loadings分析表明,5个传感器对鹿茸饮片气味特征具有较好的响应,不同规格鹿茸饮片气味差异贡献率主要体现为氮氧化物类、甲烷等短链烷烃、有机硫化物、醇醚醛酮类、芳香成分、无机硫化物等成分;PCA表明不同品种与规格鹿茸饮片,其气味的差异性比较明显;LDA发现不同品种及规格鹿茸饮片(除梅花鹿白片与粉片外)样品的气味差异均较明显,表明构成气味的物质存在差异性。结论:电子鼻技术可阐明不同品质鹿茸饮片气味的物质基础;不同品质鹿茸饮片气味存在差异性可揭示鹿茸饮片气味的科学内涵并为其质量控制提供参考。     

落花生根内生真菌Fusarium oxysporum LHS-P1-3中恩镰孢菌素类化合物及生物活性研究

目的 研究落花生根内生真菌Fusarium oxysporum LHS-P1-3的次级代谢产物及其生物活性。方法 使用硅胶色谱柱、高效液相色谱等对代谢粗提物进行分离纯化,采用质谱、核磁共振等方法测定化合物的结构,采用CCK-8法测试化合物的抗肿瘤细胞活性,使用表面等离子共振实验（surface plasmon resonance,SPR）研究化合物与胆固醇转运蛋白1（niemann-pick c1-like 1,NPC1L1）相互作用关系。结果 从落花生根内生真菌Fusarium oxysporum LHS-P1-3分离得到6个化合物,包括1个新化合物（1）与5个已知化合物：白僵菌素（2）、恩镰孢菌素I（3）、恩镰孢菌素MK1688（4）、恩镰孢菌素H（5）及恩镰孢菌素B（6）。化合物1～4对HepG2细胞和HeLa细胞的IC₅₀均小于10 μmol/L,化合物2～4则对HCT116细胞的IC₅₀小于10 μmol/L,SPR研究表明化合物2、4在20 μmol/L浓度下,可与NPC1L1蛋白相互作用。结论 化合物1为新的恩镰孢菌素类化合物,命名为恩镰孢菌素W。化合物1～5具有抗肿瘤活性,化合物2、4与NPC1L1蛋白存在相互作用,可能是NPC1L1蛋白的潜在抑制剂。     

CircCDR1as Suppresses Bone Microvascular Endothelial Cell Activity and Angiogenesis Through Targeting miR‐135b/ FIH‐1 Axis

ObjectiveThe current study investigated the role of CircCDR1as on angiogenesis of bone microvascular endothelial cells (BMECs) isolated from non‐traumatic ONFH.MethodsForty corticosteroid‐induced ONFH patients received THA were enrolled in our study. Expressions of CircCDR1as, miR‐135b, and FIH‐1 were detected by qRT‐PCR in affected necrosis tissue and non‐affected normal tissue. Bone microvascular endothelial cells (BMEC) were isolated from six patients and treated with 0.1 mg/mL hydrocortisone to establish a GC‐damaged model of BMECs. Circ CDR1as plasmid and miR‐135b mimic were transfected into BMECs. BMEC proliferation was assessed using MTT assays. The migration ability of cells was detected by scratch‐wound assays. Matrigel assay was performed to detect angiogenesis in vitro. Western blot assay was used to detect HIF‐1α, VEGF, and FIH‐1 expressions. FISH, RNA pull down, RIP, and luciferase assay were carried out to determine the interaction of CircCDR1as, miR‐135b, and FIH‐1.ResultsCircCDR1as was upregulated(2.02 ± 0.30 vs. 1.00 ± 0.10,P < 0.001) whereas miR‐135b was downregulated (0.55 ± 0.12 vs. 1.00 ± 0.10,P < 0.001) in affected tissues than in non‐affected tissues. Expression of CircCDR1as and FIH‐1 were negatively associated with miR‐135b in affected tissues (CircCDR1as with miR‐135b: r = −0.506, P < 0.001; FIH‐1 with miR‐135b r = −0.510, P < 0.001). Total blood tubule density was increased when CircCDR1as was silenced compared with NC (P < 0.01 vs. NC). The number of migrated BMECs were significantly increased in CircCDR1as silencing group compared with NC group (P < 0.05 vs. NC). In addition, CircCDR1as plasmids transfection increased the protein expressions of FIH‐1 (P < 0.05 vs. NC) and reduced the HIF‐1α as well as VEGF expression compared with NC group (P < 0.05 vs. NC). FISH, RNA pull down, RIP, and luciferase assay identified that FIH‐1 was a target of miR‐135b and could be modulated by CircCDR1as.ConclusionCircCDR1as decreases angiogenesis and proliferation of BMECs by sponging miR‐135b and upregulate FIH‐1.     

The Effect of a PEEK Material‐Based External Fixator in the Treatment of Distal Radius Fractures with Non‐Transarticular External Fixation

ObjectiveTo explore the effect of a PEEK material‐based external fixator in the treatment of distal radius fractures with non‐transarticular external fixation.MethodsThere were 48 patients in this prospective comparative study. They were divided into two groups according to the materials used: the PEEK group and the titanium group. Wrist dorsiflexion, palmar flexion, pronation, supination, radial deviation, ulnar deviation, grip strength of the palm on the affected side, kneading force, Visual Analogue Scale/Score (VAS), Disabilities of the Arm, Shoulder, and Hand (DASH) score, operation time, frequency of fluoroscopy procedures, and X‐ray results were compared between the two groups. Functional recovery was evaluated at the last follow‐up according to the wrist joint evaluation criteria.ResultsThe baseline data were comparable between the two groups, and no significant differences were found in age, sex, fracture types (P > 0.05). There was no significant difference between the two groups in the results of DASH, grip strength, and recovery of pinch force and wrist function (dorsiflexion, clavicle, ulnar deviation, deviation, pronation, and supination) (P > 0.05). Normal limb function was achieved in the two groups of patients at an average of 6 weeks after surgery, and there was no significant difference in X‐ray examination radial height (10.60 ± 1.59 vs 11.00 ± 1.53, P = 0.687), radial inclination (1.11 ± 0.24 vs 1.12 ± 0.24, P = 0.798), volar tilt (10.33 ± 2.13 vs 10.00 ± 2.08, P = 0.660), ulnar variance (20.87 ± 3.00 vs 20.38 ± 3.04, P = 0.748), and step‐off persistence (1.73 ± 0.69 vs 1.68 ± 0.72, P = 0.425) between the two groups (P > 0.05). However, the operation time (54.80 ± 12.20 vs 85.23 ± 15.14, P = 0.033) and number of fluoroscopy procedures (36.93 ± 6.89 vs 64.77 ± 9.74, P = 0.000) in the PEEK group were significantly reduced compared with those in the titanium group.ConclusionCompared with the traditional titanium external fixator, the PEEK composite external fixator has advantages, such as a shorter operation time and fewer fluoroscopy procedures when used to treat different types of distal radius fracture.     

Immunogenicity,safety and reactogenicity of ROTAVAC® in healthy infants aged 6–8 weeks in Vietnam

BackgroundROTAVAC® is derived from human 116E rotavirus (RV) neonatal strain. In this study, we evaluated the immunogenicity, safety and reactogenicity of ROTAVAC® in Vietnam.MethodWe conducted a phase IV clinical trial in healthy infants aged 6–8 weeks using the complete regimen of ROTAVAC® with three doses. Serum anti-RV IgA was measured by enzyme-linked immunosorbent assay to assess the geometric mean concentration in infants who received the complete regimen of the vaccine.ResultsA total of 360 participants were enrolled in this clinical trial. The mean age ± standard deviation at enrollment was 6.9 ± 0.6 weeks. The anti-RV IgA titer was 4.01 ± 3.74 mg/ml pre-vaccination and substantially increased to 29.27 ± 80.64 mg/ml post-vaccination. The value of logIgA significantly increased (p = 0.003) from 0.28 ± 0.79 to 1.03 ± 0.54. The proportion of participants with equal to and greater than 3-fold and 4-fold shifts in pre- to post-vaccination antibody titer (IgA) were 55.4% and 48.3%, respectively. No adverse events or serious adverse events were recorded immediately within 30 min after the administration of each dose. The most common adverse events within 14 days after each visit were fever, unusual crying and irritability. Other adverse events occurred at a low rate, and no case of intussusception was noted.ConclusionsThe complete regimen of ROTAVAC® demonstrated an immunological response with clinically acceptable safety profile. Post-completion of this study, ROTAVAC® is now a WHO-prequalified vaccine and available in Vietnam.