1.Mechanism of protection of motor neurons in spinal cord anterior horn of SNI rats by acellular nerve allografts via the Bcl-2/Cyt-C/Apaf-1 signalling pathway
Mengyuan Zheng ; Zitong Hao ; Qinghua Zhu ; Zhuangzhuang Tian ; Xingda Guo ; Yuhe Zheng ; Cheng Li ; Xiumei Fu
Acta Universitatis Medicinalis Anhui 2025;60(11):2035-2042
Objective:
To investigate the protective effects and mechanisms of acellular nerve allografts (ANA) on motor neurons in the spinal cord anterior horn of sciatic nerve injury ( SNI) rats .
Methods:
SPF grade male SD rats were randomly divided into normal , model , ANA-bridged (bridge group) , and autologous nerve transplantation groups (autograft group) , with 6 rats in each group . The SNI rat model was established using the right sciatic nerve clamp method for 10 mm . In the bridge group , the ANA was bridged to the two severed ends of the injured sciatic nerve , and in the autograft group , the autologous nerves were flipped head to tail and then bridged to the two se- vered ends . A spectrophotometer was applied to determine the DNA content in normal nerves and ANA . The foot- print test was used to determine the sciatic nerve function index (SFI) of the rats in each group , the wet weight ra- tio of the anterior tibialis muscle was calculated . The morphology and structure of the anterior horn motor neurons of the spinal cord of each group were observed by HE staining. The immunofluorescence and Western blot were used to detect Apaf-1 , Caspase-3 , Bcl-2 , Bax , and Cyt-C proteins expression in the L4-6 segment of the spinal cord .
Results:
The DNA content in the ANA prepared in this study was significantly lower than that in normal nerves (P < 0. 05) . Compared with the normal group , the SFI and wet weight ratio of the anterior tibialis muscle were re- duced in the model group (P < 0. 001) ; compared with the model group , both SFI and wet weight ratio of the ante- rior tibialis muscle significantly increased in the bridge group and the autografts group ( P < 0. 05 , P < 0. 001) , and the SFI and wet weight ratio of the anterior tibialis muscle in the autograft group were higher than those in the bridge group (P < 0. 001 , P < 0. 01) . The results of HE staining showed that the motor neurons in the anterior horn of the spinal cord of the normal group were structurally intact and had clear cytosolic boundaries; the neurons in the model group were lysed and necrotic , with blurred cytosolic boundaries; the neurons in the bridge group were less lysed and necrotic , but the nuclear translocation phenomenon could still be seen; the neurons in the autograft group were morphologically and structurally intact with clear cytosolic boundaries . Compared with the normal group , the expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins significantly increased in the model group (P < 0. 001 , P < 0. 01 , P < 0. 01 , P < 0. 05) . Compared with the model group , the expression of Apaf-1 , Caspase- 3 , Bax , and Cyt-C proteins significantly decreased (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) ; but the expres- sion of Bcl-2 protein significantly increased in the bridge group and the autograft group (P < 0. 05) . The expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins in the autografts group was lower than that in the bridge group (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) .
Conclusion
ANA can exert a protective effect on motor neurons in the anterior horn of the spinal cord of SNI rats by improving the morphology and structure of neurons , increasing the ex- pression of Bcl-2 protein , but decreasing the expression of Cyt-C , Bax , Caspase-3 , and Apaf-1 proteins in the spi- nal cord . The mechanism of ANA may be related to the Bcl-2/Cyt-C/Apaf-1-mediated mitochondrial apoptosis sig- naling pathway .
2.Applications of 3D printing technology in the treatment of mitral valve disease
LUO Xingda ; LI Xiaohui ; LIAO Shengjie ; LUO Dezhi ; YAN Xiaohui ; ZHANG Xiaoshen
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(5):509-513
Mitral valve disease is the most common cardiac valve disease. The main treatment of mitral valve disease is surgery or interventional therapy. However, as the anatomy of mitral valve is complicated, the operation is particularly difficult. As a result, it requires sophisticated experiences for surgeons. Three-dimensional (3D) printing technology can transform two-dimensional medical images into 3D solid models. So it can provide clear spatial anatomical information and offer safe and personalized treatment for the patients by simulating surgery process. This article reviews the applications of 3D printing technology in the treatment of mitral valve disease.
3.Influence of macro-pore bone block on osteoblast behavior in vitro
Tenglong HU ; Xiaojie LI ; Xiong ZHAO ; Xingda LU ; Xuguang HAO ; Bin ZHANG ; Liu YANG ; Qiang JIE
Chinese Journal of Orthopaedics 2016;(3):168-176
Objective To observe and identify the impact of a type of macro?pore bone block bioactive glass on osteo?blast in vitro. Methods Extract fluid of new bioactive glass was prepared withα?MEM culture medium as the bioactive medium group. And the concentrations of different ions were detected with Inductively Coupled Plasma?Atomic Emission Spectrometry in bioactive medium group andα?MEM medium group. MC3T3?E1 cells cultured in bioactive medium group were considered as ex?perimental group and cells cultured inα?MEM medium as control group. Giemsa and immunofluorescence staining was performed to detect the cell numbers, the karyoplasmic ratio and the average fluorescence intensity per cell. Cell proliferation and viability in different groups were detected by cell cycle analysis, MTT assay and BrdU assay, respectively. Total RNAs of cells in different groups were extracted and the expressions of ALP, OCN and collagenⅠwere measured by quantitative real time PCR. ALP stain?ing and alizarin red staining were performed to assess the differentiation and mineralization of MC3TC?E1 cells in different groups. Results The concentrations of Si and F were 40.02 ± 0.67 mg/L and 0.02 ± 0.001 mg/L in bioactive medium group, higher than 2.02±0.01 mg/L and 0.00 mg/L inα?MEM solution, and the concentration of Ca was lower than that inα?MEM solution. The con?centration of P and Na had no difference. In Giemsa staining, the cell number in 400 times field under a microscope was 106.0 ± 6.025 in bioactive medium group and 40.20 ± 3.639 inα?MEM medium group. In the immunofluorescence of vinculin, the karyo?plasmic ratio and the expression of vinculin were higher in bioactive medium group (40.85±5.720, 0.050 88±0.021 78) than inα? MEM medium group (21.93 ± 4.137, 0.023 60 ± 0.003 18). In cell cycle analysis, the proportion of cells retained in S and G2/M phase in the bioactive medium group was more than that in theα?MEM medium group after 72 hours of cell culture. In the BrdU and MTT assay, MC3T3?E1 cells in bioactive medium group both showed a higher proliferation rate with statistical significance. In MC3T3?E1 cells cultured with the bioactive medium, the expressions of osteogenesis?related genes were higher than those cultured with ordinaryα?MEM solution;in the ALP staining and alizarin red staining, the expression of ALP and the mineralization rate were higher in bioactive medium group (1.328%±0.015 36%, 2.953%±0.536 3%) than inα?MEM medium group (0.979%±0.030 59%, 1.000%±0.208 1%). Conclusion The bioactive medium promotes cell proliferation and osteoblastic differentiation of MC3T3?E1 cells, and has much more Si ions, which indicates that macro?pore bone block bioactive glass can promote cell proliferation and dif?ferentiation and has promising bioactivity.
4.Clinical Observation of Edaravone Combined with Butyl Phthalide in the Treatment of Acute Ischemic Stroke
Lian YI ; Xingda LI ; Jianxiu WANG ; Zhongling ZHANG
China Pharmacy 2016;27(29):4130-4132
OBJECTIVE:To observe clinical efficacy and safety of edaravone combined with butyl phthalide in the treatment of acute ischemic stroke (AIS). METHODS:258 AIS patients were randomly divided into observation group and control group, with 129 cases in each group. Both groups were given routine treatment as antiplatelet,improving microcirculation,controlling blood pressure,lowering blood glucose,regulating blood lipid,keeping plaque stable,nourishing brain cells. Control group was additionally given Butyl phthalide capsules orally,200 mg,tid. Observation group was additionally given Edaravone injection 30 mg added into Sodium chloride injection 100 ml,ivgtt,bid,on the basis of control group. Both groups continuously received 14 days of treatment. The serum inflammatory factor,miR-222 and neurologic impairment score of 2 groups were observed before treatment,7,14 d after treatment. Clinical efficacies and the occurrence of ADR were compared between 2 groups. RESULTS:There was no statistical significance in the serum inflammatory factor,miR-222 and neurologic impairment score between 2 groups before treatment(P>0.05). The serum inflammatory factor and neurologic impairment score of 2 groups were decreased significant-ly 7,14 d after treatment,while serum levels of miR-222 were increased significantly;the observation group was significantly bet-ter than the control group,with statistical significance(P<0.05). Total effective rate of observation group was 92.2%,which was significantly higher than that of control group (69.8%),with statistical significance (P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:Edaravone combined with butyl phthalide is effective in the treatment of AIS,and can significantly de-crease serum inflammatory factor level,promote the expression of miR-222 and improve neurologic function with good safety.


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