Loganin alleviates testicular damage and germ cell apoptosis induced by AGEs upon diabetes mellitus by suppressing the RAGE/p38MAPK/NF-κB pathway

Diabetes mellitus (DM) damages male reproduction at multiple levels, such as endocrine secretion, spermatogenesis and penile erection. We herein investigated the protective effects and mechanism of loganin targeting the advanced glycation end products (AGEs)/receptor for AGEs (RAGE)/p38 mitogen-activated protein kinase (p38MAPK)/NF-κB signalling pathway. Loganin relieved the general DM symptoms and decreased the blood glucose level of KK-Ay DM mice. Haematoxylin-eosin staining demonstrated that loganin ameliorated testicular histology and function and enhanced the activities of testis-specific markers lactate dehydrogenase (LDH), acid phosphatase (ACP) and gamma-glutamyl transferase (γ-GT). Loganin also showed evident anti-oxidative stress, anti-apoptotic and anti-inflammatory effects on DM-induced reproductive damage by restoring glutathione (GSH) level and superoxide dismutase (SOD) activity, as well as reducing reactive oxygen species (ROS) level and Bax/Bcl-2 ratio in vivo and in vitro. Western blotting exhibited that loganin significantly inhibited the AGEs/RAGE/p38MAPK/NF-κB signalling pathway. Acridine orange and ethidium bromide staining (AOEB) and Western blotting showed that loganin in combination with inhibitors of RAGE, p38MAPK and NF-κB exerted stronger anti-apoptotic effects on AGE-induced GC-2 cell damage compared with loganin alone. In conclusion, loganin can protect against DM-induced reproductive damage, probably by suppressing the AGEs/RAGE/p38MAPK/NF-κB pathway.     

Correction to: Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

Virologica Sinica - The corrected legend is given below.     

Single-tube detection of nine bacterial antibiotic-resistance genes by a 2-dimensional multiplex qPCR assay based on fluorescence and melting temperature

Molecular Biology Reports - Simple, multiplex qPCR methods are advantages for rapid molecular diagnosis of multiple antibiotics-resistant genes simultaneously. However, the number of genes can be...     

Silencing of cyt-c4 led to decrease of biofilm formation in Aeromonas hydrophila

Aquaculture suffers from a number of diseases caused by Aeromonas hydrophila. Biofilm can protect bacteria from antibiotic therapy. To identify the genes those play crucial roles in A. hydrophila biofilm formation, a library of mini-Tn10 transposon insertion mutants of A. hydrophila B11 has been constructed, and 10 mutants were subjected to biofilm formation assay. The biofilm formation ability of mutant (B188) was significantly decreased compared with B11. The DNA sequence flanking the mini-Tn10 transposon inserted showed that an ORF of approximately 576 bp of the mutant strain B188 was inserted. This ORF putatively displays the highest identity (92%) with the cytochrome c4 gene (cyt-c4) of A. hydrophila subsp. hydrophila ATCC 7966. Silencing cyt-c4 led to deficiencies in biofilm formation, adhesion, drug resistance and pathogenicity of A. hydrophila, which suggests that cyt-c4 plays crucial role in the biofilm formation and virulence mechanisms of A. hydrophila.

ABBREVIATIONS: GEN: gentamycin; SDZ: sulfadiazine; AK: amikacin; P: penicillin; CFP: cefoperazone; LEV: levofloxacin; MH: minocycline; FFC: florfenicol; TE: tetracycline; AMP: ampicillin; KAN: kanamycin; STR: streptomycin; SXT: sulfamethoxazole/trimethoprim; DO: doxycycline; OT: Oxytetracycline.     

液相质谱联用的代谢组方法优化及其在蓝细菌分析中的应用

为了准确鉴定光合蓝细菌中的各种代谢物,需要对基于液相色谱–质谱联用仪(LC-MS)的代谢组学分析方法进行有针对性的优化。本研究选取了24种涉及中心碳代谢和能量代谢的代谢物作为LC-MS的检测目标,获得了每个代谢物的最适色谱分离条件和质谱参数;同时以光合蓝细菌Synechocystis sp.PCC6803为主要对象,针对性地优化了样品前处理条件,结果显示适当延长梯度洗脱顺序表的时间并将流速设为0.2 m L/min可以得到最佳的分离效果,同时选择80%(V/V)甲醇(-80?C)作为代谢物萃取剂。分析结果证明这一代谢组分析技术可以成功地应用到光合蓝细菌的研究中。     

蚯蚓对湿地植物光合特性及净化污水能力的影响

以香蒲、芦苇和美人蕉为研究对象,并以土壤+沙子+有机质混合物为供试基质模拟人工湿地处理污水,采用向基质中加入蚯蚓与未加入蚯蚓2种处理。研究加入蚯蚓后,香蒲、芦苇和美人蕉光合速率、蒸腾速率、SPAD值和水分蒸发、蒸腾量的变化及其对净化污水能力的影响。结果表明:与未加入蚯蚓相比,加入蚯蚓后,香蒲、芦苇和美人蕉的净光合速率、蒸腾速率、SPAD值和水分蒸发、蒸腾量均增加,其中芦苇的净光合速率、蒸腾速率和水分蒸发、蒸腾量增加达到显著水平(P <0.05),而香蒲的水分蒸发、蒸腾量增加也达到显著水平(P <0.05);加入蚯蚓后,香蒲、芦苇和美人蕉对COD_Mn、NH₄⁺-N、NO₃^--N、TN和TP的去除率均增加,且香蒲和芦苇对COD_Mn的去除率显著增加 (P <0.05)。加入蚯蚓后,香蒲、芦苇和美人蕉的SPAD值均增加,说明蚯蚓能提高湿地植物对氮的吸收,增加植株中的氮含量,促进湿地植物的光合速率和蒸腾速率从而提高对污水的净化能力。     

Meta-analysis for genome-wide association study identifies multiple variants at the BIN1 locus associated with late-onset Alzheimer's disease

Recent GWAS studies focused on uncovering novel genetic loci related to AD have revealed associations with variants near CLU, CR1, PICALM and BIN1. In this study, we conducted a genome-wide association study in an independent set of 1034 cases and 1186 controls using the Illumina genotyping platforms. By coupling our data with available GWAS datasets from the ADNI and GenADA, we replicated the original associations in both PICALM (rs3851179) and CR1 (rs3818361). The PICALM variant seems to be non-significant after we adjusted for APOE e4 status. We further tested our top markers in 751 independent cases and 751 matched controls. Besides the markers close to the APOE locus, a marker (rs12989701) upstream of BIN1 locus was replicated and the combined analysis reached genome-wide significance level (p = 5E-08). We combined our data with the published Harold et al. study and meta-analysis with all available 6521 cases and 10360 controls at the BIN1 locus revealed two significant variants (rs12989701, p = 1.32E-10 and rs744373, p = 3.16E-10) in limited linkage disequilibrium (r² = 0.05) with each other. The independent contribution of both SNPs was supported by haplotype conditional analysis. We also conducted multivariate analysis in canonical pathways and identified a consistent signal in the downstream pathways targeted by Gleevec (P = 0.004 in Pfizer; P = 0.028 in ADNI and P = 0.04 in GenADA). We further tested variants in CLU, PICALM, BIN1 and CR1 for association with disease progression in 597 AD patients where longitudinal cognitive measures are sufficient. Both the PICALM and CLU variants showed nominal significant association with cognitive decline as measured by change in Clinical Dementia Rating-sum of boxes (CDR-SB) score from the baseline but did not pass multiple-test correction. Future experiments will help us better understand potential roles of these genetic loci in AD pathology.     

A Novel 2-dimensional Multiplex qPCR Assay for Single-Tube Detection of Nine Human Herpesviruses

Human herpesviruses are double-stranded DNA viruses that are classified into nine species. More than 90% of adults are ever infected with one or more herpesviruses. The symptoms of infection with different herpesviruses are diverse ranging from mild or asymptomatic infections to deadly diseases such as aggressive lymphomas and sarcomas. Timely and accurate detection of herpesvirus infection is critical for clinical management and treatment. In this study, we established a single-tube nonuple qPCR assay for detection of all nine herpesviruses using a 2-D multiplex qPCR method with a house-keeping gene as the internal control. The novel assay can detect and distinguish different herpesviruses with 30 to 300 copies per 25 μL single-tube reaction, and does not cross-react with 20 other human viruses, including DNA and RNA viruses. The robustness of the novel assay was evaluated using 170 clinical samples. The novel assay showed a high consistency (100%) with the single qPCR assay for HHVs detection. The features of simple, rapid, high sensitivity, specificity, and low cost make this assay a high potential to be widely used in clinical diagnosis and patient treatment.     

Expression of recombinant anti‐colorectal cancer large single‐chain monoclonal antibody in insect cells

Colorectal cancer is the third most commonly diagnosed cancer in the world. Monoclonal antibody (mAb) CO17‐1A recognizes the tumor‐associated antigen GA733‐2, a cell surface glycoprotein highly expressed in colorectal carcinoma cells which is applicable for preventing and curing colorectal cancer. In this study, we tried to produce a new recombinant anti‐colorectal cancer large single chain (lsc) mAb based on mAb CO17‐1A in the baculovirus‐insect cell protein expression system. Two kinds of recombinant lsc mAbs were generated where variable light chain (VL) and heavy chain (HC) of mAb CO17‐1A were fused together by an interchain linker. The only difference between the two mAbs is based on fusion of an ER retention signal (KDEL) at its C‐terminus of HC. Polymerase chain reaction analysis verified the presence of both recombinant genes in the bacmid for generating viral expression vectors in insect cells. Western blot confirmed the expression of lsc mAbs in baculovirus‐infected insect cells. Cell enzyme linked immunosorbent assay (ELISA) showed that the mAbs from cell lysates bound to SW480 and SW620 human colorectal cancer cells. These results indicate that the baculovirus insect expression system can produce anti‐colorectal lsc mAb recognizing human colorectal cancer cells.     

Monitoring leaf nitrogen accumulation in wheat with hyper-spectral remote sensing

The water-retaining capacity, percolation and evaporation of stony soil in Liupan Mountains, China, were measured in order to understand the effect of rock fragments on soil hydrological processes. The results indicated that the effective water-retaining capacity of soil is positively related with the volumetric content of rock fragments, but there is no relation between saturated water-retaining capacity and rock fragment content. For the soil layers within 0–40 cm, the steady infiltration rate increases with increasing volumetric content of rock fragments until it reaches the range of 15%–20%, and then it decreases when the rock fragment content further increases. For the soil layers below 40 cm, the steady infiltration rate always increases with increasing rock fragment content. The soil evaporation rate decreases with increasing volumetric content of rock fragments when it varies in the range of 0–20%, while the soil evaporation rate keeps basically stable when the rock fragment content is higher than 20%. The soil evaporation rate shows a rising tendency with increasing size of rock fragments.