1.Studies on in vitro capacitation of goat spermatozoa by heparin treatment.
Jia-Bo ZHOU ; Yan-Guang WU ; Li-Qing LIU ; Ming-Jiu LUO ; Zhong-Le CHANG ; Xiu-Wen TAN ; Na LIU ; Jing-He TAN
Chinese Journal of Biotechnology 2004;20(2):252-256
Systematical studies are lacking on the influencing factors and mechanisms of the heparin enhanced sperm capacitation, although many studies have shown that heparin enhanced sperm capacitation. The effect of heparin concentration and exposure time, incubation temperature and co-culture with oviductal epithelial cells or cumulus cells on goat sperm capacitation were investigated in this study. The motility, membrane and acrosome integrity and capacitated percentage of goat spermatozoa were assessed after different heparin treatments, and rates of fertilization and embryo cleavage were compared after in vitro insemination of oocytes with spermatozoa capacitated by different heparin treatments. The major results are summarized as follows: 1) When spermatozoa were capacitated with heparin at 5, 10, 25, 50 and 100 microg/mL for 45 min, 50 and 100 microg/mL heparin treatments produced the highest capacitated percentages of 55% and 56%, respectively, but the percentage of spermatozoa with intact acrosomes in the 100 microg/mL heparin treatment decreased significantly (P < 0.05) in comparison with that in the control group, indicating that the optimal heparin concentration for goat sperm capacitation would be 50 microg/mL. 2) Capacitated percentage of spermatozoa increased with extension of treatment time when goat sperm were treated with 50 microg/mL heparin for 0, 10, 20, 30, 45, 60 or 120 min. Although heparin treatments for 45 to 120 min did not differ significantly (P > 0.05) in capacitated sperm percentages, sperm motility and membrane integrity decreased significantly when treated with heparin for 120 min. This suggested that the optimal exposure time of heparin at 50 microg/mL for goat sperm capacitation would be 45 to 60 min. 3) Significantly higher capacitated percentages of spermatozoa were obtained when goat sperm were treated at 42 and 38.5 degrees C than at 15 and 37 degrees C, but sperm motility and acrosome integrity were significantly lower when spermatozoa were treated at 42 degrees C than they were treated at other temperatures. Temperature of 38.5 degrees C would, therefore, be the optimal temperature for goat sperm capacitation. 4) The capacitated percentage of spermatozoa was significantly higher when goat sperm were co-cultured with oviductal epithelial cells than when treated with heparin alone or co-cultured with cumulus cells, but sperm motility and membrane and acrosome integrity did not differ significantly among the three treatments. Rates of fertilization (91.3%) and cleavage (72.2%) were significantly higher in the oviductal epithelial cell co-culture group than those in the heparin alone group. This indicated that co-culture with oviductal epithelial cells significantly enhanced goat sperm capacitation by heparin treatment.
Acrosome Reaction
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drug effects
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physiology
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Animals
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Coculture Techniques
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Epithelial Cells
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cytology
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Fallopian Tubes
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cytology
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Female
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Fertilization in Vitro
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Goats
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Heparin
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pharmacology
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Male
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Sperm Capacitation
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drug effects
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physiology
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Sperm Motility
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Spermatozoa
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cytology
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physiology
2.Effect of minimally invasive Ivor-Lewis esophagectomy on acute phase responses in patients with esophageal carcinoma.
Qiang FANG ; Yong-tao HAN ; Guang-guo REN ; Chang-ning YE ; Tai-chang TAN ; Lin PENG ; Bo XIAO ; Wen-guang XIAO
Chinese Journal of Oncology 2010;32(11):868-871
OBJECTIVETo explore the effect of minimally invasive Ivor-Lewis esophagectomy on acute phase responses in patients with esophageal carcinoma.
METHODSForty-eight patients with middle or low thoracic esophageal carcinoma underwent Ivor-Lewis esophagectomy. The patients were divided into small incision group (n = 25) and conventional group (n = 23) according to the patients' will. Serum levels of acute phase proteins C reactive protein (CRP), haptoglobin (HPT), α₁-acid glycoprotein (α₁-AG), ceruloplasmin (CER), transferrin (TRF), β₂-microglobulin (β₂-MG), album protein (ALB), interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were measured and compared on 1st day before operation, at 18 hours as well as 3rd and 7th day after operation.
RESULTSThere was no significant difference in all the acute phase proteins indicators and IL-6 between the small incision and conventional groups at each time points after operation (P > 0.05). In both groups the levels of CRP, α₁-AG and HPT were significantly higher after operation than before operation (P < 0.05). The levels of ALB and TRF were significantly lower after operation than before operation (P < 0.05). The levels of CER and β₂-MG were not significantly different during perioperative period (P > 0.05). The level of TNF-α was significantly higher in the small incision group than that in the conventional group at the 18 hours postoperationally (P < 0.05), and were not significantly different on the other time points between the two groups (P > 0.05).
CONCLUSIONCompared with conventional operation, the small incision Ivor-Lewis esophagectomy do not significantly alleviate the stress of the surgical trauma in patients. Unchanging the essence of operation, if one is trying to minimize the stress caused by surgery on patients, the key factor is not the size of incision. An effective approach should be found in other operation-related factors.
Acute-Phase Proteins ; metabolism ; Aged ; C-Reactive Protein ; metabolism ; Carcinoma, Squamous Cell ; blood ; surgery ; Ceruloplasmin ; metabolism ; Esophageal Neoplasms ; blood ; surgery ; Esophagectomy ; methods ; Female ; Haptoglobins ; metabolism ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Orosomucoid ; metabolism ; Perioperative Period ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Transferrin ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; beta 2-Microglobulin ; blood
3. Polarization of M1 tumor associated macrophage promoted by the activation of TLR3 signal pathway
Bo LIU ; Xia WANG ; Tai-Zhong CHEN ; Guang-Liang LI ; Chang-Chun TAN ; Yong CHEN ; Shao-Qiang DUAN
Asian Pacific Journal of Tropical Medicine 2016;9(5):484-488
Objective To investigate the correlation between activation of toll-like receptors 3 (TLR3) signaling pathway and tumor-associated macrophage and its effect on the tumor growth. Methods The mice Lewis lung cancer cell lines 3LL and melanoma B16H10 were used to construct the subcutaneous transplantation tumor models and then they were treated with Poly-ICLC. The curative effect was observed and then the T cell and macrophage phenotypes infiltrated in local tumor were detected by flow cytometry. After the in vitro culture of mouse bone marrow-derived macrophage, the real-time PCR and western blot were applied to detect the expression of macrophage activation markers and the activation of intracellular signaling pathways. Results The survival time of mice with brown tumor treated with Poly-ICLC significantly increased and the tumor growth was inhibited. The ratio of local tumor-infiltrated Treg decreased, while the ratio of CD8