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1.
National Journal of Andrology ; (12): 584-590, 2016.
Article in Chinese | WPRIM | ID: wpr-262350

ABSTRACT

<p><b>Objective</b>To study the expression of human lysozyme-like protein 6 (LYZL6) in the male reproductive system and its physiological role.</p><p><b>METHODS</b>The recombinant P. pastoris strain was cultured and induced with methanol to express LYZL6, followed by purification using chitin affinity chromatography. The bactericidal activity of the recombinant LYZL6 was observed by bilayer agar plate diffusion assay, and then the recombinant protein was used as an immunogen to generate polyclonal antibodies, whose specificity was examined by ELISA. The distribution of LYZL6 in the human tissue and semen was identified by Western blotting and the subcellular localization in the testis was investigated by immunohistochemistry.</p><p><b>RESULTS</b>At pH 5.6, recombinant LYZL6 exhibited a high bacteriolytic activity against M. lysodeikticus. ELISA analysis showed that the anti-LYZL6 polyclonal antibodies could bind the recombinant protein with a high specificity. Western blot manifested the expression of LYZL6 in the testis and epididymis, higher in the former than in the latter. LYZL6 was also detected in the sperm protein extract, while protein bands were not observed in the seminal plasma. Immunodetection with a specific antiserum localized the LYZL6 protein in the late spermatocytes and round spermatids.</p><p><b>CONCLUSIONS</b>LYZL6 has a higher bacteriolytic activity under low pH condition and is bound to spermatozoa after secreted in the testicular epithelia, suggesting that LYZL6 could act as a potential hydrolase for carbohydrates in zona pellucida penetration.</p>


Subject(s)
Humans , Male , Blotting, Western , Epididymis , Metabolism , Muramidase , Genetics , Metabolism , Pichia , Metabolism , Recombinant Proteins , Genetics , Metabolism , Semen , Metabolism , Spermatozoa , Metabolism , Testis , Metabolism
2.
Acta Physiologica Sinica ; (6): 503-508, 2004.
Article in English | WPRIM | ID: wpr-352743

ABSTRACT

Experiments were performed on male Sprague-Dawley (SD) rats anesthetized with a mixture of urethane and chloralose. A rat model of acute myocardial ischemia (AMI) was made by ligation of the left anterior descending branch of the coronary artery (LAD). After the LAD ligation, the ischemia area of the left ventricular wall became somewhat pale immediately. Under a light microscope, the pathological examination revealed that all the cells were swollen and in red color when the cardiac section was stained with hematoxylin basic fuchsin picric acid (HBFP), which indicated a typical change in the myocardial ischemia. In the AMI model, it was found that cardiac functions were markedly attenuated, such as decreases in the heart rate (HR), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), maximal rate for left ventricular pressure rising and declining (+/-dp/dt(max)), velocity of contractile element (V(CE)) and total area of cardiac force loop (L(0)), and an increase in the left ventricular end diastolic pressure (LVEDP). In such AMI rats, application of electroacupuncture (EA) at "Neiguan" acupoints (Pe 6) for 20 min could obviously improve the above-mentioned cardiac functions. After microinjection of nitro-L-arginine (L-NNA), an inhibitor of nitric oxide synthase (NOS), was made into the rostral ventrolateral medulla (RVLM), the curative effect of EA on myocardial ischemia was reduced significantly or abolished, while after microinjection of normal saline of the same volume was made into the RVLM, the improving effect of EA remained. These results suggest that the effect of EA on myocardial ischemia is possibly mediated by the nitric oxide (NO) in the RVLM.


Subject(s)
Animals , Male , Rats , Acupuncture Points , Electroacupuncture , Medulla Oblongata , Metabolism , Myocardial Ischemia , Metabolism , Nitric Oxide , Physiology , Rats, Sprague-Dawley , Ventricular Function
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