1.Impact of Vascular Injury on Neointimal Hyperplasia in Experimental Rabbit Model
Jixiang YUAN ; Liang ZHANG ; Shujun WEI
Chinese Circulation Journal 2016;31(12):1210-1214
Objective: To study the impact of vascular injury on neointimal hyperplasia and the expressions of nuclear transcription factor-κb (NF-κb), tissue factor (TF), endothelin-1 (ET-1) and matrix metalloproteinase-3 (MMP-3) in experimental rabbit model.
Methods:A total of 32 male New Zealand big-eared white rabbits were used to establish vascular injury model by femoral artery puncture, balloon was sent to abdominal aorta via the wire followed by balloon dilatation to strain abdominal aorta. Blood sample was taken from femoral artery sheath 1 minute after operation, and the rabbits were killed at 4 weeks after operation, meanwhile blood sample was taken from the heart. Injured arteries were isolated, ifxed and embedded; slices were stained by HE, basic fuchsine and NF-κb immunohistochemical methods for light microscope observation; slices were also stained by lead and uranium for transmission electron microscope observation. Neointimal thickness was measured by computer analysis, vascular injury integral and NF-κb positive cell rate were determined, blood levels of TF, ET-1 and MMP-3 were examined by ELISA. The relationship between vascular injury integral and the contents of TF, ET-1, MMP-3 and NF-κb positive cell rate were analyzed by SPSS statistical software.
Results: The optimal iftting curve between vascular injury integral and neointimal thickness was S-shaped cubic curve. NF-κb was mainly expressed in neointima, vascular injury integral was positively related to NF-κb positive cell percentage, Pearson correlation coeffcient was 0.916,P<0.05. Vascular injury integral was positively related to the contents of 1 min post-operative TF; 1 min and 4 weeks post-operative ET-1; 1 min and 4 weeks post-operative MMP-3; Pearson correlation coeffcients were 0.925, 0.957, 0.947, 0.914 and 0.940 respectively, allP<0.05.
Conclusion: Vascular injury may activate NF-κb pathway, promote ET-1 and MMP-3 expression, therefore accelerating neointimal hyperplasia, leading negative vascular remodeling, TF was an initiating factor for neointimal hyperplasia. Internal elastic lamina was the key structure reflecting vascular injury, it is the only barrier hindering neointimal hyperplasia in experimental rabbit model.
2.Intracranial aneurysm isolation combined with extra-intracranial bypass in treatment of middle cerebral artery aneurysm
Li LI ; Desheng WEI ; Shujun LI
Journal of Regional Anatomy and Operative Surgery 2016;25(11):801-803
Objective To investigate the effect of intracranial aneurysm isolation combined with extra-intracranial bypass in treatment of middle cerebral artery aneurysm.Methods From January 2013 to March 2015,there were 32 patients with complex middle cerebral artery aneurysm in our hospital,who were treated by intracranial aneurysm isolation combined with extra-intracranial bypass.The clinical symptoms and image examination after surgery were retrospectively analyzed,and the hemodynamic index of middle cerebral artery before and after treat-ment was statistically analyzed.Results The postoperative clinical symptoms of 25 patients were relieved,the mRS score was 0 point.Seven patients received surgery,1 week later,the muscle force of involved side was grade 2 to 3,the mRS score was 2 to 3 point,the blood flowing smoothly in target vessel,with no original aneurysm,no new ischemic stroke or infarct.After intervention,systolic and diastolic blood flow ve-locity were faster than those before the intervention(P <0.05),blood flow resistance were lower than those before intervention(P <0.05). Conclusion The intracranial aneurysm isolation combined with extra-intracranial bypass can effectively improve the clinical symptoms of pa-tients with complex intracranial aneurysms,with high graft patency rate and reliable effect.
3.Preparing acellular nerve allografts by combined freeze-thaw and chemical methods
Shujun GUAN ; Wei WANG ; Yan LI
Chinese Journal of Tissue Engineering Research 2015;(12):1914-1918
BACKGROUND:Host immune rejection is the main problem for nerve alograft in the repair of nerve defects. Therefore, how to avoid and minimize the immune rejection is the key to the success of nerve alografting. OBJECTIVE: To develop a new nerve pretreatment method by which Schwann cels and myelin can be removed from the peripheral nerve of dogs while the basilar membrane can be reserved integraly in order to obtain acelular nerve alografts. METHODS:Bilateral sciatic nerves from healthy adult dogs were taken and pretreated with the combined freeze-thaw and chemical methods folowed by microscopic observation of ultrastructural features, histological staining and western blot analysis of its ingredients. RESULTS AND CONCLUSION:Pretreated acelular nerves with good ductility and excelent epineurium toughness were empty basal lamina tubes with no Schwann cels, myelin and fragments that were al removed thoroughly, but the basilar membrane was fuly retained. These findings indicate that the optimized combination of freeze-thaw and chemical methods can efficiently clear Schwann cels and myelin which are the major antigenic components in the peripheral nerve, while preserve the basilar membrane to promote nerve regeneration. Therefore, this method can be an ideal method for preparation of tissue engineered nerves.
4.Estrogen and Ability of Learning and Memory:A Review
Wei DONG ; Wei NING ; Cuizhen ZHANG ; Shujun TIAN
Chinese Journal of Rehabilitation Theory and Practice 2008;14(9):814-816
Estrogen is an important active endogenous substance which is needed by female to sustain normal psychological and physiological function.Studies have demonstrated that estrogen has an extensive effect on the central nervous system.This review explained the relation between estrogen and learning on a few aspects such as neurobiochemistry,neurophysiology,neuroanatomy,signal transduction and so on.
5.Effect of mechanical lesion in medial region of nucleus retrofacialis on respiratory rhythm
Min HE ; Li LI ; Jun LI ; Shujun WEI
Chinese Journal of Tissue Engineering Research 2005;9(21):219-221
BACKGROUND: Until now, the definite source of respiratory rhythm generation and the ontogenic mechanism are still in controversy. Previous our investigating indicated that respiration would stop due to the gentle press on the hypoglossal root under the surface of ventral medulla of rabbit. OBJECTIVE: To observe the influence of mechanical lesion in medial re gion of nucleus retrofacialis on respiratory rhythm of a living rabbit model.DESIGN: Randomized controlled study.SETTING: At the Physiology Teaching Room of Kunming Medical College.MATERIALS: This study was carried out at the Physiology Teaching Room of Kunming Medical College from February 2002 to December 2002. Totally 54 healthy rabbits were randomly selected, amongst which 22 rabbits were submitted to the mechanical damage of the medial region of bilateral nucleus retrofacialis by the dorsal processes, and 18 received the same damage by the ventral process, 14 rabbits used for observing the changes of blood pressure andheart rate due to the mechanical damage.METHODS: The ventral and dorsal medullar of rabbits were exposed and the bilateral vagotomy at the cervical level were cut. Discharge of the phrenic nerve was used as the respiratory index. Stainless pipe of 0. 8 mm or 0.4 mm diameter was used to make mechanical damage at the medial region in order to investigate the changes of respiration. Fourteen rabbits were selected and submitted to the bilateral mechanical damage, the changes of blood pressure and heart rate were observed at post-traumatic 1 -3 minutes.MAIN OUTCOME MEASURES: ① The changes of respiratory rhythm induced by the mechanical damage of the medial region of bilateral nucleus retrofacialis. ② The number of phrenic nerve discharge and respiratory frequency. ③ The changes of blood pressure and heart rate at post-lesion 1-3minutes.RESULTS: Data of 54 rabbits were analyzed. ① Irreversible disappearance of respiratory rhythm was found in 12 out of 18 rabbits (the positive rate was 67% ) due to the mechanical damage by the ventral processes, comparing to 3 / 14 (14%) by the dorsal process. ② Rhythmic discharge of phrenic nerve became scarce and irregular in all rabbits who has irreversible disappearance of respiratory rhythm, the inspiratory and expiratory discharging phase were significantly prolonged, moreover respiratory rhythm would disappear if expiratory phase was extremely extended, respiration stopped at expiratory phase. ③ When the medial region of bilateral nucleus retrofacialis was partially destroyed, the still existed respiration of 27 rabbits was found immediately inhibited, phrenic nerve discharge frequency obviously decreased [ (43.5 ± 6.4)%, P < 0. 001 ], and its discharge number was significantly decreased [ (42.0 ± 3.7 )%, P < 0. 001 ], moreover the inspiratory and expiratory discharging phase were found obviously extended, with expiratory phase was in particular. ④ From the results of histological examination on the above mechanical lesion. Respiration stop or obvious attenuation was found mostly due to the damage at the medial region of nucleus retrofacialis.⑤ The blood pressure and heart rate changed slightly by 5% -7% at post-traumatic 1 -3 minutes, there was not significantly different from the normal value ( P > 0. 05).CONCLUSION: The medial region of nucleus retrofacialis might be the main source of respiratory rhythm generation, and the related neuron within it would be the important component involving in the generation of respiratory rhythm.
6.Crystallography, morphology, and thermal properties of starch in Fritillaria thunbergii and F. ussurensis as well as comparison with potato starch
Shujun WANG ; Wenyuan GAO ; Wei JIA ; Peigen XIAO
Chinese Traditional and Herbal Drugs 2005;36(8):1216-1220
Objective To fully understand the medicinal plants of Fritillaria L. , the physicochemical properties of starch in two species of Fritillaria L. , F. thunbergii and F. ussurensis. were investigated by means of various analytical methods. Methods The properties of starch in the two different species of Fritillaria L. were compared by X-ray diffraction, scanning electron microscope (SEM) and themogravimetric analysis (TGA). Results The crystal type of starch in the two species of Fritillaria L.was the characteristic B-type which was in consistent with that of potato starch. The degrees of crystallinity of F. thunbergii starch and F. ussurensis starch were about 29.9% and 20.1%, respectively. However,the degree of crystallinity of the potato starch was 44.9%. From the crystallinity degree of the starch in two species of Fritillaria L. , it could be concluded that the content of amylose in F. ussurensis starch was higher than that in F. thunbergii starch. The granule size of the starch in two species of Fritillaria L.ranged from 5 to 40 μm, which were all smaller than that of the potato. The starch granule in two species of Fritillaria L. was in cycloidal or elliptic-shape. It could be concluded that the thermal stability of the starch in two species of Fritillaria L. was different due to the different structures of different starch in various plants by TGA. Conclusion The physicochemical properties of starch in two different species of Fritillaria L. differ a lot due to their geographical origin.
7.Relationship between nitric oxide in cervical microenvironment and different HPV types and effect on cervical cancer cells
Xuemin WEI ; Qing WANG ; Shujun GAO ; Long SUI
Chinese Journal of Obstetrics and Gynecology 2011;46(4):260-265
Objective To study the relationship between nitric oxide within cervical microenvironment and different HPV types as well as the effect of sodium nitroprusside( SNP), a nitric oxide donor, on the proliferation and apoptosis of cervical cancer cell lines. Methods HPV typing test was assessed from 115 women by using high-risk HPV (HR-HPV) 21 typing test and the release of cervical nitric oxide(NO) was assessed as nitrate, nitrite in cervical fluid. Cervical NO was then compared between women showing different HPV types. Proliferation of Caski and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell apoptosis was detected by flow cytometry after 24 hours treated by different final concentration of SNP (0. 125,0. 25,0. 5,1.0 and 2. 0 mmol/L, respectively). The expressions of HPV E6,E7 gene mRNA and p53 protein were detected by SYBR Green Ⅰ quantitative real-time PCR and western blot. Results ( 1 ) The cervical NO release of women with HR-HPV was higher compared to that in HPV negative women [ (47. 6 ± 1.4) μmol/L vs ( 22. 8 ± 0. 3 ) μmol/L; P < 0. 05 ]; but there was no statistical difference between low-risk HPV (LR-HPV) group [ (24. 1 ± 1.2 ) μmol/L] and control group (P >0. 05 ). (2)After 24 hours treated by different final concentration of SNP, the results shown that SNP could inhibited the proliferation and increased apoptosis rate in Caski and HeLa cells, in which the concentration of SNP ≥ 1.0 mmol/L , there were significantly different ( P < 0. 05 ), while when SNP ≥2. 0mmol/L, the proliferation of cells inhibited seriously. Treated by SNP ( 1.0 mmol/L ) 24 hours, the expressions of HPV18 E6, E7 mRNA in HeLa cells were reduced from 27. 362 ±0. 191,22. 962 ±0. 053 to19. 181 ±0. 360, 17. 571 ±0. 010 and the protein expression of p53 increased from 1. 17 ±0. 03 to 0. 23 ±0. 05, there were statistically significant differences between adding SNP group and the control group ( P <0. 05); but there were no statistically significant differences in HPV16 E6, E7 mRNA and that of p53 in Caski cells( P > 0. 05 ). Conclusions The presence of HR-HPV is associated with an increased release of NO in the human uterine cervix; NO could inhibit the growth and proliferation and enhance the apoptosis of cervical cancer cells, inhibit the expression of HPV18 E6, E7 mRNA in HeLa cells and activate the expression of p53 protein, the mechanism may be due to higher sensitivity of HeLa cells (cervical adenocarcinoma cell) to SNP. The increasing release of NO may play a role in regulating the elimination of HPV in cervical microenvironment, which is a part of mucous membrane immunity.
8.The research on the different expression of matrix metalloproteinase-9 in cervical columnar ectopy and cervical intraepithelial neoplasia
Wei YUAN ; Shujun CAO ; Liping GAO ; Daohua YANG ; Chengmin QIU
Chinese Journal of Postgraduates of Medicine 2014;37(24):12-14
Objective To explore the different probability of carcinoma and the necessity of treatment in cervical columnar ectopy (CCE),normal smooth cervix and cervical intraepithelial neoplasia(CIN) by investigating the different expression of matrix metalloproteinase-9(MMP-9).Methods The expression of MMP-9 was detected by using immunohistochemical method in 30 cases of CCE,28 cases of CIN and 30 cases of normal smooth cervix from 2007 to 2012.Results The positive expression of MMP-9protein in CIN was 96.4% (27/28),CCE was 23.3% (7/30) and normal smooth cervix was 20.0% (6/30)respectively.The positive expression of MMP-9 protein was higher in CIN than CCE and normal smooth cervix,the difference has statistical significance (P< 0.01),there was no significant difference between CCE and normal smooth cervix (P > 0.05).Conclusions CCE,which was a physiological performance of cervical erosion,should not be overtreated.But the women of childbearing age should be routinely performed in cervical cancer screening.
9.Effect of ulinastatin on tumor necrosis factor and interleukin-6 in rats with radiation-induced lung injury
Pengtao BAO ; Haowen QI ; Wei GAO ; Shujun LI ; Lijiang ZHANG
Chinese Journal of Radiological Medicine and Protection 2009;29(2):154-157
Objective To observe the effect of ulinastatin on tumor necrosis factor(TNF-α)and interleukin-6(IL-6)in radiation-induced lung injury.Methods Severity-two female SD rats were randomly divided into 3 groups as control group,irradiation group and treatment group(administered with Ulinastatin).Rats in irradiation group and treatment group were irradiated with linear accelerator at a single dose of 25 Gy.After irradiation rats in treatment group were injected daily with ulinastatin at a dose of 100000 U-kg-1·d-1 for 7 days through caudal vein while rats in control group and irradiation group were injected with the same volume of saline.Rats were killed at 2 h,4,8 and 24 weeks.Samples of lung tissues were observed by using HE staining.Expression of TNF-α in lung was determined by Western blot and expression of IL-6 in serum was determined by ELISA.Data were analyzed by SPSS software.Results Expressions of TNF-α in lung and IL-6 in serum increased significantly after irradiated in irradiation group compared with control group,and it reached the peak at 4 weeks(q=5.63、6.21,P<0.01).Though expressions of TNF-α and IL-6 in ffeatment group also increased compared with control group,the difference between irradiation group and treatment group was statistic significantly(q=4.97、7.42,P<0.01).Conclusions TNF-α and IL-6 play an important role in radiation-induced lung injury.Ulinastatin could suppress the inflammatory response and radiation-induced lung injury effectively by decreasing the levels of TNF-α and IL-6.
10.Estrogen Increased ERK1/2 Phosphorylation Levels in Hippocampal CA4 Region of Ovariectomized Rats
Mingchun JIANG ; Ning ZOU ; Wei NING ; Shujun TIAN
Chinese Journal of Rehabilitation Theory and Practice 2007;13(3):229-232,封3
Objective To investigate the effects of estrogen on extracellular signal-regulated kinase-1/2(ERK1/2) phosphorylation levels in hippocampal CA4 region of ovariectomized rats. Methods Experimental female Wistar rats were randomly divided into three groups: intact (INT) group, ovariectomized (OVX) group, and OVX+estrogen (1 mg/kg, weekly) group. Radioimmunoassay (RIA) was used to measure the estrogen content in plasma, and the levels of ERK1/2 phosphorylation were measured with immunohistochemistry. Results Compared with the control animals, the plasma estrogen level was significantly lower in OVX group (P<0.001). However, the plasma estrogen level was higher in the OVX+estrogen group than that of the OVX group (P<0.001). Immunohistochemical staining showed that the phospho-ERK1/2 (p-ERK1/2) substances were present in the nucleus and cytosol of neurons. Compared with the control group, immunostaining positive cells markedly decreased in OVX group, with light nucleus and cytosol staining in the hippocampal CA4 region (P<0.001). The phospho-ERK1/2 positive neurons increased more obviously in OVX+estrogen group than in the OVX group (P<0.001). Conclusion Estrogen can increase ERK1/2 phosphorylation levels in hippocampi of aging female rats, which suggests that the ERK signal transduction pathway may be regulated by estrogen during the process of aging.