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中国生物医学工程学会
北京玛格泰克科技发展有限公司
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2023 Vol. 32, No. 1
Published: 2023-03-30
Research papers
Research papers
1
Phospholipase C-Zeta Reveals the Underlying Pathological Influence after Density Gradient Centrifugation, Microfluidic Sorting, and Cryopreservation of Human Sperm
CAI Jing-jing
Objective:
Sperm preparation techniques and cryopreservation are widely used in assisted reproductive techniques (ART). How to improve the quality of sperm management is a matter of great concern. Phospholipase C-zeta (PLCζ) is considered a sperm-specific agent that activates oocyte activation and thus playing a crucial role in male fertility. However, the potential mechanisms by which semen processing and cryopreservation on PLCζ contribute to keyhole have not been addressed.
Methods:
In this study, semen samples were taken from have not been addressed 10 normozoospermic men. Each semen sample was assigned to the following groups: density gradient centrifugation (DGC) as control, microfluidic sorting, and cryopreservation. Sperm parameters of molity, viability, membrane integrity, and intracellular ROS were evaluated during sperm preparation and cryopreservation. The expression of PLCζ in human sperm was determined by immunofluorescence and western blotting.
Results:
The results showed that molity, viability, and membrane integrity decreased in cryopreservation group. Intracellular ROS were also significantly increased compared to the the control group. There was no significant difference between DGC and microfluidic sorting group. Our investigation revealed that total levels of PLCζ were comparable between DGC and microfluidic sorting, but there were significantly reduced levels of PLCζ after cryopreservation as quantified by both immunofluorescenceand immunoblotting. PLCζ immunofluorescence in sperm revealed different PLCζ localization patterns around the acrosomal(Ac), equatorial (Eq), post-acrosomal (PA) areas of sperm heads, and their combination. The predominant patterns of PLCζ localization in DGC were similar to that of microfluidic sorting, with strong, with staining. In contrast, PLCζ staining in freeze-thawed sperm was considerably weaker fluorescence intensity.
Conclusion:
This study clarified the mechanism of sperm preparation and cryopreservation underlying effect on sperm characteristic, accompanied with PLCζ expresion. We demonstrated that microfluidic sorting provides a highly efficient preparation method for clinical selection of PLCζ-expressing sperm comparable to DGC gene expression. It is suggested that the cryopreservation of sperm has a significant detrimental effect on PLCζ.
2023 Vol. 32 (1): 1-10 [
Abstract
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31
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11
Effect of Pressure-Controlled Inverse Ratio Ventilation on WBC, PCT and CRP in Rats Undergoing Radical Gastrectomy and Related Mechanism
LI Chun-yang, PENG Xiao-hui, WAN Hui-lai, ZHENG Yue
Objective:
To explore the effect of pressure-controlled inverse ratio ventilation on partial pressure of oxygen (PaO
2
), partial pressure of carbon dioxide (PaCO
2
), dynamic lung compliance (Cdyn), white blood cell (WBC), procalcitonin (PCT), C-reactive protein (CRP), and other indicators.
Methods:
A total of 100 healthy Wistar rats, aged 6-8 weeks and weighing 300-356 g, were used to establish gastric cancer models with 1-methyl-3-nitro-1-nitrosoguanidine (MNNG). 60 gastric cancer rats were randomly divided into three groups and treated with laparoscopic radical gastrectomy after anesthesia. Rats in the VCV group were given volume control ventilation, rats in the PCV group were given pressure control ventilation, and rats in the PC-IRV group were given pressure control inverse ratio ventilation. The levels of WBC, PCT and CRP in the three groups were observed and compared on the 1st, 3rd and 5th day after operation.
Results:
There were significant differences in PaO
2
levels at 15 min after ventilation mode change and at the time of pneumoperitoneum closure, and in PaCO
2
and Cdyn levels at 10 min after induction and at 15 min after ventilation mode change among the three groups (
P
<0.05). The PaO
2
levels in PC-IRV group were higher than those in VCV group and PCV group at 15 min after the ventilation mode was changed and at the time of closing the pneumoperitoneum. The level of PaCO
2
in PC-IRV group was lower than that in VCV group and PCV group 10 min after induction and 15 min after ventilation mode change, and the level of PaCO
2
in PC-IRV group was lower than that in VCV group when pneumoperitoneum was turned off. The level of Cdyn in PC-IRV group was higher than that in VCV group and PCV group at 10 min after induction and 15 min after ventilation mode change. The level of Cdyn in PC-IRV group was higher than that in PCV group at the time of closing pneumoperitoneum. The Cdyn level in PCV group was lower than that in VCV group at 15 min after ventilation mode change. There were significant differences in WBC levels among the three groups on the 1st day after operation (
P
<0.05), and there were no significant differences in PCT and CRP levels among the three groups on the 1st day after operation (
P
>0.05).There were significant differences in WBC, PCT and CRP levels in the three groups of rats on the 3rd and 5th days after surgery (
P
<0.05). Except for PCT and CRP on the 1st day after operation, the WBC on the 1st, 3rd and 5th days after operation, PCT and CRP on the 3rd and 5th days after operation in PCV group and PC-IRV group were lower than those in VCV group (
P
<0.05). Except for WBC, PCT and CRP on 1st day after operation, the levels of WBC, PCT and CRP on the 3rd and 5th days after operation in PC-IRV group were lower than those in PCV group (
P
<0.05).
Conclusion:
During laparoscopic radical gastrectomy for gastric cancer, pressure-controlled inverse ratio ventilation can help rats maintain good respiratory status and lung compliance during the operation, which can effectively alleviate the postoperative inflammatory response.
2023 Vol. 32 (1): 11-18 [
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13
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19
Study on Basic Biomarkers of Alzheimer's Disease with Depression
SHI Lei, LIN Duo-duo, WANG Wei-bing, WU Hai-long
Objective:
To establish a rat model of Alzheimer's disease with depression, and to explore the common serum indicators that can be used to assess the occurrence of depression in Alzheimer's disease patients by routine blood testing.
Methods:
120 healthy male SD rats were randomly divided into healthy group, control group, dementia group and dementia with depression group, with 30 rats in each group. The rats in the three groups were tested for the basic biological markers by means of routine serum monitoring.
Results:
30 rats in each healthy group and control group, 28 rats in dementia group and 27 rats in dementia with depression group completed the experiment. There were statistical differences in the levels of each biomarker in the four groups of rats (
P
<0.05); there was no statistical difference in the levels of each biomarker between the healthy group and the control group (
P
>0.05); The levels of serum IL-1β, IL-6, TNF-α, CRP, and Hcy in the dementia group were higher than those in the healthy group and the control group, and the 5-HT levels were lower than those in the healthy group and the control group (
P
<0.05); The levels of IL-1β, IL-6, TNF-α, CRP, and Hcy in the depression group were higher than those in the other three groups, and the levels of 5-HT were lower than those in the other three groups (
P
>0.05).
Conclusion:
The occurrence of depressive symptoms in Alzheimer's rats can be predicted to some extent by routine serum IL-1β, IL-6, TNF-α, CRP, Hcy and 5-HT levels, and it is inferred that the above indicators can be used as important biological markers to assess the occurrence of depression in Alzheimer's patients.
2023 Vol. 32 (1): 19-24 [
Abstract
] (
15
)
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25
Study on the Relationship Between the Growth and Development Speed and the Expression of GH and IGF-1 in Children
QUE Yong-xiang, ZHANG Ling-zhao, ZHANG Sheng-can, YOU Qiu-xia
Objective:
To investigate the relationship between the growth and development speed and the expression of growth hormone(GH) and insulin-like growth factor-1(IGF-1) in children.
Methods:
A total of 78 healthy children who received physical examination in our hospital from December 2020 to December 2022 were randomly selected by cluster sampling, which were divided into three groups according to the standard deviation unit curve of height and weight, namely, the delayed group (n=11), the normal group (n=51) and the tachycardia group. Fasting venous blood was collected in the early morning to detect the levels of GH and IGF-1 from the three groups of children according to the relevant operation standards. Pearson correlation analysis was performed between Δheight, Δweight and GH, IGF-1 in all children.
Results:
Overall analysis showed that there were significant differences in height and weight among the three groups of children on the day of enrollment, at 3, 6, 9 and 12 months after enrollment (
P
<0.05). Intra-group comparison showed that the height and weight levels of the children in the three groups at 12 months were significantly higher than those on the day of enrollment, and the difference was statistically significant (
P
<0.05). The height and weight of children in normal group and tachycardia group were significantly higher than those in tardy group on the day of enrollment, at 3, 6, 9 and 12 months after enrollment (
P
<0.05). The height and weight of the children in the tachycardia group were higher than those in the normal group on the day of admission, at 3, 6, 9 and 12 months after admission (
P
<0.05). There were significant differences in Δheight and Δweight among the three groups (
P
<0.05). The Δheight and Δweight of children in normal group and tachycardia group were higher than those in delayed group, and the Δheight and Δweight of children in tachycardia group were higher than those in normal group (
P
<0.05). There were significant differences in the levels of GH and IGF-1 among the three groups on the day of enrollment and 12 months after enrollment (
P
<0.05). The levels of GH and IGF-1 in normal group and tachycardia group were higher than those in delayed group on the same day and 12 months after treatment (
P
<0.05), and the levels of GH and IGF-1 in tachycardia group were higher than those in normal group on the same day and 12 months after treatment (
P
<0.05). The changes of GH and IGF0-1 in the three groups were compared, and there was no significant difference in ΔGH and ΔIGF-1 among the three groups (
P
<0.05). Pearson correlation analysis showed that Δheight and Δweight were positively correlated with GH and IGF-1 (R=0.715-0.886,
P
<0.05).
Conclusion:
There is a close relationship between the growth and development speed of children and the expression of GH and IGF-1 in the body, which can be used as a diagnostic basis for the diseases related to abnormal growth and development in children.
2023 Vol. 32 (1): 25-31 [
Abstract
] (
13
)
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32
Study on the Significance of TLR-9 Expression in Rats with Acute Lung Injury Induced by Thoracic Trauma
SHI Yu-zhen, LI Yong-shun, LIN Sheng-yang, CAO Cheng-zhang
Objective:
To investigate the significance of toll receptor-9 (TLR-9) expression in rats with acute lung injury (ALI) induced by thoracic trauma.
Methods:
Fifty healthy male SD rats were selected for the study, and 40 of them were randomly selected to establish the acute lung injury model induced by thoracic trauma, and 8 rats were randomly selected at 5, 24, 48 and 72 h after the impact, which were killed by cervical dislocation after arterial bleeding, and the lung tissue sections were made. Randomly select 8 rats from the remaining 10 rats as the control group, take blood and kill them to make lung tissue sections for reference. The lung dry/wet weight ratio (W/D) of the five groups were compared, and the levels of TLR-9 in bronchoalveolar lavage fluid (BALF) and serum, and the levels of TLR-9 mRNA, IL-6 mRNA, TLR-9 protein and IL-6 protein in lung tissue were detected.
Results:
There was no significant difference in W/D levels among the control group, experimental group 1 and experimental group 4 (
P
>0.05), while the W/D level of experimental group 3 was higher than that of the other four groups, and the W/D level of experimental group 2 was higher than that of the control group (
P
<0.05). The levels of ILR-9 in serum and BALF of rats in experimental group 1, experimental group 2 and experimental group 4 had no significant difference (
P
>0.05), which were higher than those in control group and lower than those in experimental group 3 (
P
<0.05). The levels of TLR-9 mRNA, IL-6 mRNA, TLR-9 protein and IL-6 protein were the highest in the experimental group 3, and the lowest in the control group (
P
<0.05). The levels of TLR-9 mRNA and IL-6 mRNA in lung tissue of rats in experimental group 1, experimental group 2 and experimental group 4 had no significant difference (
P
>0.05). There was no significant difference in the levels of TLR-9 protein and IL-6 protein between experimental group 2 and experimental group 4 (
P
>0.05), which were higher than those in experimental group 1 (
P
<0.05).
Conclusion:
The expression level of TLR-9 is significantly increased in rats after thoracic trauma, and the expression of TLR-9 can reflect the degree of lung injury in rats after trauma to a certain extent, which is inferred to be related to the inflammatory response after lung injury.
2023 Vol. 32 (1): 32-38 [
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39
Effect of Atorvastatin on the Expression of Inflammatory Cytokines in Rats with Klebsiella Pneumonia
QUE Wei-wei, LI You-tang, WENG Duan-li
Objective:
To explore the effect of atorvastatin on the expression of inflammatory cytokines in rats with Klebsiella pneumonia.
Methods:
90 healthy SPF-grade SD rats were randomly divided into three groups: atorvastatin group, model group and blank group (with 10 rats in each group), 30 rats in each treatment period (3,6,9 d). A rat model of Klebsiella pneumonia was constructed, in which the blank group and the model group were given the same volume of saline, while the atorvastatin group was given 10 ml/kg of atorvastatin by intraperitoneal instillation. The rats were killed on the 10th day after administration, and the lung tissue was extracted to detect the pathological results and the expression of inflammatory cytokines was detected in serum.
Results:
Lung histopathology showed that lung histopathology and fibrosis were improved in atorvastatin group, and alveolar structure integrity ≥50% and collagen fiber precipitation ≤10% in atorvastatin group indicated that the model was successful. The expression of inflammatory cytokines showed that the levels of IL-6, TNF-α and TGF-β in the atorvastatin group and the model group were significantly increased compared with the blank group, with statistically significant differences(
P
<0.05). The levels of IL-6, TNF-α and TGF-β in atorvastatin group were lower than those in model group, and the levels of IL-10 in atorvastatin group were higher than those in model group, with statistically significant differences (
P
<0.05). After 9 days, the levels of IL-6, TNF-α, TGF-β and IL-10 in the atorvastatin group and the model group were higher than those in the blank group, with statistically significant differences (
P
<0.05). The results of immunological function study showed that WBC, RBC and PLT in orvastatin group and model group were significantly reduced at 3 d compared with the blank group, with statistically significant differences(
P
<0.05). After 6 and 9 d, WBC, RBC and PLT in atorvastatin group and model group were lower than those in blank group, and WBC, RBC and PLT in atorvastatin group were lower than those in model group, with statistically significant differences(
P
<0.05).
Conclusion:
Atorvastatin can significantly improve the immune dysfunction and the expression of inflammatory cytokines in rats with Klebsiella pneumonia.
2023 Vol. 32 (1): 39-46 [
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12
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