PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distribu...PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distributed specifically in testis and is highly immunogenic, function of which relates to energy metabolism and capacitation of the sperm. Therefore, in this study, we observed the effect of purified native PH-20 and LDH-C4 antigens on sperm function and morphology in dog. Native PH-20 (46, 32 kDa) and LDH-C4 (36, 30, 28) sub units, purified from dog sperm SDS- extracts were used to test as immunocontraceptive candidates in stray dog. Dogs 1, 2, 3, 4, 5/6 and 7/8 were immunized with 46, 32, 36, 30/28, 46/32, 36/30/28 kDa sub units respectively by i/m route and semen was analysed at weekly intervals. High titre of 3200 - 6400 was achieved in all dogs after three boosters. Immune response in treated dogs was confirmed by increase in Ig level in blood serum and reaction of partially purified serum Ig of immunized dogs with respective antigens in double immune diffusion and immunoblots. Immunization of dogs either with individual PH-20/ LDHC sub units or pooled PH-20/LDHC4 sub units showed a significant effect on sperm count, percent motility, viability, HOST and sperm morphology. All the sperm parameters declined to a significant level (p < 0.05) between 45 - 115 DPI and remained low than that of the recommended values for fertile dog semen till 175 DPI. The incubation of spermatozoa in HOS solution in the presence of anti-PH-20 and anti-LDHC Ig significantly reduced the percent HOS positive spermatozoa. A significant decline (p ≤ 0.05) in acrylamide penetration assay was also observed in the presence of anti 46 and 30/28 kDa Ig. Since percent motility, sperm count, HOST and percent normal spermatozoa in immunized dogs were significantly less than that of recommended fertile dog semen, it can be concluded that PH-20 (46/32 kDa) and LDHC4 (36/30/28 kDa) sub units had an immunocontraceptive potential in dog.展开更多
One of the distinct hallmarks of cancer cells is aerobic glycolysis(Warburg effect). Lactate dehydrogenase A(LDHA) is thought to play a key role in aerobic glycolysis and has been extensively studied, while lactate de...One of the distinct hallmarks of cancer cells is aerobic glycolysis(Warburg effect). Lactate dehydrogenase A(LDHA) is thought to play a key role in aerobic glycolysis and has been extensively studied, while lactate dehydrogenase C(LDHC), an isoform of LDHA, has received much less attention.Here we showed that human LDHC was significantly expressed in lung cancer tissues, overexpression of Ldhc in mice could promote tumor growth, and knock-down of LDHC could inhibit the proliferation of lung cancer A549 cells. We solved the first crystal structure of human LDHC4 and found that the activesite loop of LDHC4 adopted a distinct conformation compared to LDHA4 and lactate dehydrogenase B4(LDHB4). Moreover, we found that(ethylamino)(oxo)acetic acid shows about 10 times selective inhibition against LDHC4 over LDHA4 and LDHB4. Our studies suggest that LDHC4 is a potential target for anticancer drug discovery and(ethylamino)(oxo)acetic acid provides a good start to develop lead compounds for selective drugs targeting LDHC4.展开更多
文摘PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distributed specifically in testis and is highly immunogenic, function of which relates to energy metabolism and capacitation of the sperm. Therefore, in this study, we observed the effect of purified native PH-20 and LDH-C4 antigens on sperm function and morphology in dog. Native PH-20 (46, 32 kDa) and LDH-C4 (36, 30, 28) sub units, purified from dog sperm SDS- extracts were used to test as immunocontraceptive candidates in stray dog. Dogs 1, 2, 3, 4, 5/6 and 7/8 were immunized with 46, 32, 36, 30/28, 46/32, 36/30/28 kDa sub units respectively by i/m route and semen was analysed at weekly intervals. High titre of 3200 - 6400 was achieved in all dogs after three boosters. Immune response in treated dogs was confirmed by increase in Ig level in blood serum and reaction of partially purified serum Ig of immunized dogs with respective antigens in double immune diffusion and immunoblots. Immunization of dogs either with individual PH-20/ LDHC sub units or pooled PH-20/LDHC4 sub units showed a significant effect on sperm count, percent motility, viability, HOST and sperm morphology. All the sperm parameters declined to a significant level (p < 0.05) between 45 - 115 DPI and remained low than that of the recommended values for fertile dog semen till 175 DPI. The incubation of spermatozoa in HOS solution in the presence of anti-PH-20 and anti-LDHC Ig significantly reduced the percent HOS positive spermatozoa. A significant decline (p ≤ 0.05) in acrylamide penetration assay was also observed in the presence of anti 46 and 30/28 kDa Ig. Since percent motility, sperm count, HOST and percent normal spermatozoa in immunized dogs were significantly less than that of recommended fertile dog semen, it can be concluded that PH-20 (46/32 kDa) and LDHC4 (36/30/28 kDa) sub units had an immunocontraceptive potential in dog.
基金National Natural Science Foundation of China (81602653)the Fundamental Research Funds for the Central Universities,Southwest Minzu University (2020NYBPY13,China)。
文摘One of the distinct hallmarks of cancer cells is aerobic glycolysis(Warburg effect). Lactate dehydrogenase A(LDHA) is thought to play a key role in aerobic glycolysis and has been extensively studied, while lactate dehydrogenase C(LDHC), an isoform of LDHA, has received much less attention.Here we showed that human LDHC was significantly expressed in lung cancer tissues, overexpression of Ldhc in mice could promote tumor growth, and knock-down of LDHC could inhibit the proliferation of lung cancer A549 cells. We solved the first crystal structure of human LDHC4 and found that the activesite loop of LDHC4 adopted a distinct conformation compared to LDHA4 and lactate dehydrogenase B4(LDHB4). Moreover, we found that(ethylamino)(oxo)acetic acid shows about 10 times selective inhibition against LDHC4 over LDHA4 and LDHB4. Our studies suggest that LDHC4 is a potential target for anticancer drug discovery and(ethylamino)(oxo)acetic acid provides a good start to develop lead compounds for selective drugs targeting LDHC4.
