Shenling Baizhusan Improves Spermatogenesis in Hyperuricemia Oligoasthenospermia Mice by Regulating Nrf2/ARE Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Shenling Baizhusan on the treatment of oligoasthenospermia with hyperuricemia （HUA）. MethodThirty-two male Kunming （KM） mice were randomly divided into blank group （n=6）， model group （n=6）， high-dose Shenling Baizhusan group （n=7）， low-dose Shenling Baizhusan group （n=7）， and febuxostat group （n=6）. Except for the blank group， all other groups received intraperitoneal injection of potassium oxazinate suspension （600 mg·kg-1） for 7 days. After modeling， the high-dose Shenling Baizhusan group and the low-dose Shenling Baizhusan group were orally administered with 20.14 g·kg-1 and 10.07 g·kg-1 of Shenling Baizhusan， respectively. The Febuxostat group was orally administered with 0.25 g·kg-1 of Febuxostat， while the blank group and model group were orally administered with the same volume of physiological saline. Oral administration was performed once a day for 14 consecutive days， after which samples were collected. Biochemical methods were used to measure serum uric acid （UA）， superoxide dismutase （SOD） and malondialdehyde （MDA） in testicular tissue. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in testicular tissue and evaluate the spermatogenesis function. Automated sperm analyzer was used to measure sperm density and motility. Single-cell gel electrophoresis （SCGE） was used to assess sperm DNA integrity. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） was used to detect testicular cell apoptosis rate. Western blot analysis was performed to measure the protein expression levels of Kelch-like ECH-associated protein 1 （Keap1）， nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and Caspase-3 in testicular tissue. Real-time polymerase chain reaction （PCR） was conducted to evaluate the mRNA expression levels of Keap1， Nrf2， and HO-1 in testicular tissue. ResultCompared with the blank group， the model group showed elevated serum UA level （P<0.01）， decreased testicular spermatogenesis function， sperm density， and motility （P<0.01）， and increased sperm trailing rate and testicular cell apoptosis rate （P<0.01）. Compared with the model group， the high-dose Shenling Baizhusan group showed significant improvements in the above-mentioned indicators （P<0.05， P<0.01）. Additionally， the expression levels of Keap1， Bax， and Caspase-3 in testicular tissue were reduced， while the expression levels of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01）. The mRNA level of Keap1 decreased （P<0.05， P<0.01）， while the mRNA levels of Nrf2 and HO-1 increased （P<0.05， P<0.01）. ConclusionShenling Baizhusan can significantly improve HUA oligoasthenospermia， and its mechanism may be related to the Nrf2/antioxidant response element （ARE） signaling pathway.

Identification and verification of key salt-tolerant amino acid sites of banana MaNHX5 / 生物工程学报

In order to improve the salt tolerance of banana NHX genes, we cloned a MaNHX5 gene from Musa acuminata L. AAA group and predicted the key salt-tolerant amino acid sites and mutant protein structure changes of MaNHX5 by using bioinformatics tools. The 276-position serine (S) of MaNHX5 protein was successfully mutated to aspartic acid (D) by site-directed mutagenesis, and the AXT3 salt-sensitive mutant yeast was used for a functional complementation test. The results showed that after the mutated MaNHX5 gene was transferred to AXT3 salt-sensitive mutant yeast, the salt tolerance of the mutant yeast was significantly improved under 200 mmol/L NaCl treatment. It is hypothesized that Ser276 of MaNHX5 protein plays an important role in the transport of Na+ across the tonoplast.

Immunosensor construction of label free detection of CD4+T lymphocytes / 国际检验医学杂志

Objective To construct an immunosensor for detecting CD4+ T lymphocytes without labeling .Methods The staphy-lococcus protein A(SPA) method was adopted to conduct the oriented immobilization of CD4 monoclonal antibodies on the gold in-terdigitated microelectrode surface for capturing CD4+ T lymphocytes .Then cyclic voltammetry(CV) method was used to conduct the representation of modification situation on the gold interdigitated microelectrode surface .Finally the electrochemical impedance spectroscopy(EIS) was used to detect the impedance of CD4+ T lymphocytes captured by the immunosensor .The standard curve was drawn by the impedance values change obtained by the equivalent electric circuit fitting .Results The linear range of this im-munosensor for detecting CD4+ T lymphocytes was (5 × 103 -5 .0 × 106 )/mL ,with lower detection limit of 5 .0 × 102/mL .Conclu-sion The constructed immunosensor has accurate and reliable detection results uhidn is simple to operate accurate ,convenient and cheap ,which might be expected to be used in the real-time detection system ,and offers help for realizing rapid ,accurate and inex-pensive CD4+ T lymphocyte count .

A comparative study on three methods of enzyme linked immunosorbent assay for detecting hepatitis B virus Pre S1 and S2 antigen / 国际检验医学杂志

Objective To compare the three different methods of enzyme linked immunosorbent assay(ELISA),to select the best method for clinical diagnosis and treatment.Methods Addcare ELISA800,TECAN freedom evolyzer and manual ELISA method were used to detect hepatitis B virus Pre S1 antigen(preS1Ag) hepatitis B virus Pre S2 antigen(preS2Ag) in confrontation control product samples and serum specimens from patients with HBV,and the results were analyzed by statistical methods.Results The batch precisions of the three methods to detect pre-S1Ag were 4.73%,5.38%,11.87%,the batch precisions of the three methods to detect pre-S2Ag were 4.91%,5.04%,11.75%.The inter batch precisions of the three methods to detect pre-S1Ag were 6.63%,7.90%,13.26%,the inter batch precisions of the three methods to detect pre-S2Ag were 6.74%,7.81%,12.59%.All the sensitivities were 100.00%.Conclusion All the three methods have good consistency,which could be used in the detection of Pre-S1Ag and Pre-S2Ag.The precision of Addcare ELISA800 is the best,which could further improve the quality of clinical testing.

Recent Progress of Colorimetric Assays Based on Gold Nanoparticles for Biomolecules / 分析化学

Biochemical analysis assays based on colorimetric methods using gold nanoparticles have many advantages including high sensitivity, good selectivity, naked-eyes readout and complex instruments free. These methods have good prospects in applications. The biomolecule assay is highly relative with human health. This review mainly focuses on colorimetric assays applying gold nanoparticles for biomolecules detection.

Methods for Cell Micropatterning on Two-Dimensional Surfaces and Their Applications in Biology / 分析化学

The technologies that we call cell micropatterning allow the control of the shape and size of cell adhesion. Combination of micro/nano technology, surface chemistry, electrochemistry and photochemistry enables us to control the adhesion, migration, differentiation of cells and the interactions between different types of cells. These methodologies bring about a new platform for the studies of cell biology. A number of techniques for cell patterning and compares their advantages and disadvantages were reviewed in this article. The applications of cell micropatterning, including those for fundamental studies in cell biology, tissue engineering and cell-based biosensors were also discussed.