食用菌超细粉免疫调节和抗氧化功能研究

以糙皮侧耳、双孢蘑菇、金针菇、3个黑木耳品种、3个香菇品种为材料,通过测定小鼠体重、免疫器官重量、细胞免疫功能、体液免疫功能、单核-巨噬细胞的吞噬功能、血清及肝脏中谷胱甘肽过氧化物酶、超氧化物歧化酶、肝脏中白介素-6、丙二醛等指标进行小鼠免疫功能的评价研究。结果表明:"四川青川"香菇、"北神奇1号"黑木耳和"黑龙江黑29"黑木耳能显著增强小鼠的细胞免疫功能;糙皮侧耳、"辽宁恒仁"香菇、金针菇、"北神奇1号"黑木耳、"黑龙江黑29"黑木耳均能显著促进小鼠单核吞噬细胞的吞噬能力,增强小鼠机体的免疫力;糙皮侧耳、"四川青川"香菇、"辽宁恒仁"香菇、金针菇和"黑龙江黑29"黑木耳具有提高小鼠肝脏中白介素-6含量的作用;双孢蘑菇、"福建古田"香菇、"辽宁恒仁"香菇、金针菇增加小鼠血清溶血素水平,具有显著的特异性体液免疫增强作用。此外,双孢蘑菇、金针菇、"北神奇1号"黑木耳、"吉林黑29"黑木耳均可显著提高血清和肝脏中的GSH-PX活性;糙皮侧耳能提高小鼠血清中和肝脏中SOD活性;从而达到清除自由基和抗氧化损伤的作用。综合上述结果,9种食用菌超细粉均具有显著增强小鼠免疫调节能力和抗氧化作用,为食用菌功能食品产品开发利用提供支撑。     

The Role of China in the Global Spread of the Current Cholera Pandemic

Epidemics and pandemics of cholera, a severe diarrheal disease, have occurred since the early 19th century and waves of epidemic disease continue today. Cholera epidemics are caused by individual, genetically monomorphic lineages of Vibrio cholerae: the ongoing seventh pandemic, which has spread globally since 1961, is associated with lineage L2 of biotype El Tor. Previous genomic studies of the epidemiology of the seventh pandemic identified three successive sub-lineages within L2, designated waves 1 to 3, which spread globally from the Bay of Bengal on multiple occasions. However, these studies did not include samples from China, which also experienced multiple epidemics of cholera in recent decades. We sequenced the genomes of 71 strains isolated in China between 1961 and 2010, as well as eight from other sources, and compared them with 181 published genomes. The results indicated that outbreaks in China between 1960 and 1990 were associated with wave 1 whereas later outbreaks were associated with wave 2. However, the previously defined waves overlapped temporally, and are an inadequate representation of the shape of the global genealogy. We therefore suggest replacing them by a series of tightly delineated clades. Between 1960 and 1990 multiple such clades were imported into China, underwent further microevolution there and then spread to other countries. China was thus both a sink and source during the pandemic spread of V. cholerae, and needs to be included in reconstructions of the global patterns of spread of cholera.     

Tenotomy or Tenodesis for the Long Head of Biceps Lesions in Shoulders: A Systematic Review and Meta-Analysis

Background

Both tenotomy and tenodesis have been widely used for the treatment of long head of biceps tendon (LHBT) lesions, but the optimal strategy remains considerably controversial. In this meta-analysis of published studies, we compared the results of the two procedures.

Methods

A literature search that compared tenotomy with tenodesis was performed using MEDLINE, and Embase until August 2014. A total of 7 studies reporting data on 622 subjects were included. Study quality was evaluated using the PEDro critical appraisal tool and the NO quality assessment tool.

Results

Data synthesis showed higher functional outcomes, a lower complication rate, and longer surgical time in patients managed with tenodesis compared to tenotomy (Constant score, P = 0.02; Popeye sign, P < 0.001; cramp pain, P = 0.04; surgical time, P < 0.001, respectively).

Conclusion

This meta-analysis indicates that tenodesis results in better arm function and lower incidences of cramp pain and Popeye sign in LHBT lesions, while the procedure required longer surgical time compared to tenotomy. More sufficiently powered studies would be required to further determine the optimal strategy.     

Efficacy of a Parainfluenza Virus 5 (PIV5)-Based H7N9 Vaccine in Mice and Guinea Pigs: Antibody Titer towards HA Was Not a Good Indicator for Protection

H7N9 has caused fatal infections in humans. A safe and effective vaccine is the best way to prevent large-scale outbreaks in the human population. Parainfluenza virus 5 (PIV5), an avirulent paramyxovirus, is a promising vaccine vector. In this work, we generated a recombinant PIV5 expressing the HA gene of H7N9 (PIV5-H7) and tested its efficacy against infection with influenza virus A/Anhui/1/2013 (H7N9) in mice and guinea pigs. PIV5-H7 protected the mice against lethal H7N9 challenge. Interestingly, the protection did not require antibody since PIV5-H7 protected JhD mice that do not produce antibody against lethal H7N9 challenge. Furthermore, transfer of anti-H7 serum did not protect mice against H7N9 challenge. PIV5-H7 generated high HAI titers in guinea pigs, however it did not protect against H7N9 infection or transmission. Intriguingly, immunization of guinea pigs with PIV5-H7 and PIV5 expressing NP of influenza A virus H5N1 (PIV5-NP) conferred protection against H7N9 infection and transmission. Thus, we have obtained a H7N9 vaccine that protected both mice and guinea pigs against lethal H7N9 challenge and infection respectively.     

小鼠胚胎体外发育培养基中氨基酸含量变化

通过检测哺乳动物早期胚胎体外发育过程中可以消耗或生成某些氨基酸的含量,可以了解胚胎的发育潜能。利用反相高效液相色谱法(RP-HPLC)检测KSOMaa培养基中17种氨基酸含量的变化,了解昆明小白鼠(Mus musculus)植入前胚胎体外培养过程中氨基酸含量的变化,旨在寻找一种能有效支持昆明小鼠胚胎体外发育的培养基氨基酸组成,优化小鼠胚胎体外培养体系。将180枚原核胚分为9组,体外培养至囊胚,分别于胚胎发育不同时期取样做高效液相色谱分析。这些氨基酸在胚胎发育不同时期的培养基中含量变化可分为5种类型:①在2细胞期增加但在4细胞期、8～16细胞期减少,囊胚期又增加的氨基酸(甘氨酸、亮氨酸、苏氨酸、缬氨酸、苯丙氨酸、酪氨酸);②在胚胎发育各个时期均下降(谷氨酸、甲硫氨酸、精氨酸、组氨酸);③在胚胎发育各个时期均增加(丝氨酸、赖氨酸、丙氨酸);④2细胞期含量减少而在其他时期持续增加(天冬氨酸、脯氨酸、色氨酸);⑤囊胚期减少,其他时期都有增加(异亮氨酸)。     

Receptor‐like kinase OsSIK1 improves drought and salt stress tolerance in rice (Oryza sativa) plants

Receptor‐like kinases (RLKs) play essential roles in plant growth, development and responses to environmental stresses. A putative RLK gene, OsSIK1, with extracellular leucine‐rich repeats was cloned and characterized in rice (Oryza sativa). OsSIK1 exhibits kinase activity in the presence of Mn²⁺, and the OsSIK1 kinase domain has the ability to autophosphorylate and phosphorylate myelin basic protein (MBP). OsSIK1 promoter‐GUS analysis revealed that OsSIK1 is expressed mainly in the stem and spikelet in rice. The expression of OsSIK1 is mainly induced by salt, drought and H₂O₂ treatments. Transgenic rice plants with overexpression of OsSIK1 show higher tolerance to salt and drought stresses than control plants. On the contrary, the knock‐out mutants sik1‐1 and sik1‐2, as well as RNA interference (RNAi) plants, are sensitive to drought and salt stresses. The activities of peroxidase, superoxide dismutase and catalase are enhanced significantly in OsSIK1‐overexpressing plants. Also, the accumulation of H₂O₂ in leaves of OsSIK1‐overexpressing plants is much less than that of the mutants, RNAi plants and control plants, as measured by 3,3′‐diamino benzidine (DAB) staining. We also show that OsSIK1 affects stomatal density in the abaxial and adaxial leaf epidermis of rice. These results indicate that OsSIK1 plays important roles in salt and drought stress tolerance in rice, through the activation of the antioxidative system.     

Optimization of Pulsed-Field Gel Electrophoresis for Legionella pneumophila Subtyping

A total of 32 strains of Legionella pneumophila were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of L. pneumophila. Twenty-six isolates of L. pneumophila with various origins and 11 isolates from five different water systems were used as the panels. For optimization of electrophoretic parameters (EPs) of SfiI PFGE, 26 isolates were analyzed with SfiI digestion, using four EPs yielding the same D value. The EP of a switch time of 5 to 50 s for 21 h had the smallest similarity coefficients and was declared the optimal EP for SfiI PFGE of L. pneumophila. By software analysis and pilot study, AscI was chosen as another PFGE enzyme. AscI PFGE could cluster the isolates from each water system into the same or very similar patterns and had a high degree of typing concordance with other molecular methods. In evaluating the discriminatory power of AscI with the panel of 26 isolates, AscI PFGE gave one single pattern and a D value of 100%. AscI PFGE had a high discriminatory power and a high degree of consistency with epidemiological data and other molecular typing methods for L. pneumophila subtyping, and hence, AscI could be used as a restriction enzyme in PFGE subtyping of L. pneumophila.Legionella pneumophila is an environmental organism that can cause disease in humans and is increasingly recognized as an important pathogen causing nosocomial pneumonia. Potable water systems (14, 26), spa water (28), and cooling towers (7, 13) are among the sources implicated in outbreaks of Legionnaires’ disease. Transmission of bacteria from the environment to humans occurs via inhalation or aspiration of Legionella-containing aerosols (3, 5). Strain differentiation is necessary for the identification of sources of contamination and determination of routes of transmission; this could in turn enable us to more accurately detect outbreaks and limit the spread of L. pneumophila infections. A variety of subtyping techniques have been used to identify and characterize L. pneumophila strains, including monoclonal antibody (MAb) analysis (16, 19), ribotyping (4), amplified fragment length polymorphism (AFLP) analysis (9, 22), PCR-based methods (15, 24), sequence-based typing (SBT) (9, 16), and pulsed-field gel electrophoresis (PFGE) (1, 6).Preliminary reports demonstrated that PFGE is a highly discriminative epidemiological marker for subtyping of L. pneumophila (6, 11, 23, 25), and a number of L. pneumophila PFGE protocols have been described in the literature (1, 2, 4, 14); however, most laboratories that use PFGE to subtype L. pneumophila cannot compare their results because the protocols differ from each other in critical parameters, such as the restriction enzymes and electrophoresis conditions used to generate the DNA fingerprints. To enhance our ability to monitor this pathogen, there is an urgent need for a standardized L. pneumophila PFGE protocol which can readily be implemented in different laboratories for information interpretation.An optimal PFGE protocol produces a suitable number of restriction fragments and gives distinct patterns by agarose gel electrophoresis, with these determined by the restriction enzymes and the electrophoretic parameters (EPs) used. SfiI is the most frequently used enzyme in conventional PFGE protocols for L. pneumophila, and there are several different EPs for SfiI digestion used by investigators for characterization and epidemiological studies. For a certain restriction enzyme, selection of the EP with the smallest similarity coefficients will increase the discriminatory power of PFGE. As the first phase of this study, we compared the similarity coefficients obtained for four EPs with SfiI digestion and determined the one with the maximal discriminatory power.There were some problems found in practical applications of epidemiological investigation of L. pneumophila by PFGE with single SfiI digestion, such as having epidemiologically unrelated strains exhibit the same patterns (30) and the appearance of “ghost” or “phantom” bands. Combination use of two enzymes would give a higher discriminatory power and more accurate results (10, 29). Thus, as the second phase of this study, we selected another suitable enzyme and compared it with SfiI to evaluate the possibility of its use in characterization and epidemiological studies of L. pneumophila.     

The acute phase response stimulates the expression of angiopoietin like protein 4

The acute phase response is characterized by elevations in serum triglyceride levels due to both an increase in hepatic VLDL production and a delay in the clearance of triglyceride rich lipoproteins secondary to a decrease in lipoprotein lipase (LPL) activity. Recently there has been a marked increase in our understanding of factors that regulate LPL activity. GPIHBP1 facilitates the interaction of LPL and lipoproteins thereby allowing lipolysis to occur. Angiopoietin like proteins (ANGPTL) 3 and 4 inhibit LPL activity. In the present study, treatment of mice with LPS, an activator of TLR4 and a model of Gram-negative infections, did not alter the expression of GPIHBP1 in heart or adipose tissue. However, LPS decreased the expression of ANGPTL3 in liver and increased the expression of ANGPTL4 in heart, muscle, and adipose tissue. Serum ANGPTL4 protein levels were markedly increased at 8 and 16 h following LPS treatment. Administration of zymosan, an activator of TLR2 and a model of fungal infections, also increased serum ANGPTL4 protein and mRNA levels in liver, heart, muscle, and adipose tissue. Finally, treatment of 3T3-L1 adipocytes with LPS or cytokines (TNF alpha, IL-1 beta, and interferon gamma) stimulated ANGPTL4 expression. These studies demonstrate that ANGPTL4 is a positive acute phase protein and the increase in ANGPTL4 could contribute to the hypertriglyceridemia that characteristically occurs during the acute phase response by inhibiting LPL activity.     

renalase在人近曲肾小管上皮细胞系的表达与分泌

目的:探讨renalase在人近曲肾小管上皮细胞系(HK-2)的表达与分泌,为进一步研究细胞水平renalase及其通路建立稳定的实验平台。方法:以HK-2细胞系作为研究材料。①应用Westernblot方法检测renalase蛋白的表达。②用real-timePCR方法检测renalasemRNA表达的变化。③用ELISA方法检测细胞上清液中renalase的浓度。结果:在mRNA水平及蛋白水平均检测到renalase表达。结论:首次在mRNA水平及蛋白水平证实了HK-2细胞能够表达renalase,为进一步研究儿茶酚胺或缺血缺氧刺激下细胞renalase的表达奠定了基础。