1.Growth of Schwann cells in silk fibroin scaffolds with different pore sizes
Wei JIAO ; Xia ZHAO ; Yan LU ; Yang WANG ; Jianchuan WEN ; Zhengzhong SHAO
Chinese Journal of Tissue Engineering Research 2011;15(25):4607-4610
BACKGROUND: The growth behaviors of cells on the biomaterials scaffold may be affected by the topography, pore size, wettability, porosity and other factors.OBJECTIVE: This research is aimed to observe the growth and proliferation of Schwann cells in silk fibroin scaffolds with different pore sizes. METHODS: Two kinds of silk fibroin scaffolds with different pore sizes were prepared, including a large pore size scaffold (pore size 50-60 μm) and a small pore size scaffold (pore size 10-20 μm). Schwann cells (R3 [33-10ras3]) served as seed cells and incubated in 37 ℃, 5% CO2 incubation box. When cells filled up the culture bottle bottom and formed a dense monolayer, they were digested and the cell concentration adjusted, then Schwann cells were seeded onto the surface of the porous silk fibroin scaffolds with different pore sizes. After seven days of co-culture, the growth and proliferation of Schwann cells were observed under scanning electron microscope.RESULTS AND CONCLUSION: The growth of Schwann cells on the surface of silk fibroin scaffolds with different pore sizes was varied. On the surface of small pore size scaffold (10-20 μm), the cell density was low, while the phenotype of cells was bipolar, cells arranged in parallel or linked as the cell chains. On the surface of large pore size scaffold (50-60 μm), more cells could be seen, but most of the cells were in the shape of single sphere, cells clustered on the surface of the porous scaffold or aggregated as a bunch of grape at the bottom of pores. Only few cells were bipolar and lied on the ridge between the pores. The result showed that the pore size of porous silk fibroin scaffolds is an influential factor for the growth and adhesion of Schwann cells. Schwann cells are conducive to grow on the scaffolds with pore size larger than cell body diameter.
2.Chronic hepatic injury modeling in mice induced by D-galactosamine and lipopolysaccharide combination
Yanan ZHAI ; Jingjing WANG ; Meng LI ; Yafei CHI ; Xia MENG ; Boya ENG ; Kun JIAO ; Jing LU
Chinese Journal of Comparative Medicine 2014;(5):62-65
Objective To research the method of Chronic hepatic injury modeling in mice induced by D -galactosamine and lipopolysaccharide combination . Methods Injected D-galactosamine ( 30 mg/mL ) and lipopolysaccharide ( 2μg/mL ) combination by intraperitoneal injection , two days at a time for 8 weeks .Monitored variation of diet and weight; detected serum level of alanine aminotransferase ( ALT ) and aspartate aminotransferase (AST), been put to death in mice and removed the liver tissue .strained hepatic tissue by the HE and Masoon dye to observe Liver tissue structure and cellular morphology and the degree of fibrosis .Results Lipopolysaccharide and D-galactosamine combination resulted in ALT rise , hepatocyte degeneration and necrosis ,collagen fiber hyperplasia obviously . Conclusion D-galactosamine and Lipopolysaccharide combination could induce mice chronic hepatic injury modeling .
3.Cloning and expression of recombinant BmkNaTx12, a new voltage-gated sodium channel from scorpion Buthus martensii Karsch
Yang XIA ; Lingling HU ; Jiao CHEN ; Wuguang LU ; Huaijiang YAN ; Jiege HUO ; Peng CAO
Journal of China Pharmaceutical University 2017;48(2):220-226
A cDNA library was constructed from glands of wild Buthus martensii Karsch scorpion.A new sodium channel long-chain toxin BmkNaTx12 was identified by sequence alignment,and its full-length cDNA sequence was cloned.Sequence alignment showed that the sequence of BmkNaTx12 had high structural similarity with the β-toxin Cn12 from Mexican Centruroides noxius scorpion.The predicted structure of BmkNaTx12 was obtained by homologous model construction and the highest scoring model was selected.The protein structure was found to be stable at 300 ns by molecular dynamics optimization,and fluctuations at amino acids 20,35,and 52 are most like-ly due to the location of these amino acids in the loop region.Then,an expression system of recombinant BmkNaTx12-Fc fusion protein was constructed.The optimal expression time of recombinant protein was determined by Western blot and the fusion protein was successfully expressed in HEK293 cells.The purity of recombinant fusion protein which was obtained by protein A affinity chromatography was up to 95% by SDS-PAGE gel electrophoresis and HPLC.In addition,the whole-cell patch-clamp assay results suggest that 1 μmoL/L BmkNaTx12-Fc can increase 20% Nav1.7 peak current.These results laid a foundation for the further study of biological function.
4.Changes of glucagon and somatostatin levels during oral glucose tolerance test in patients with type 2 diabetes mellitus
Limin MA ; Fangzhen XIA ; Hui ZHU ; Yi CHEN ; Jiao YU ; Ying LI ; Yanxiang LI ; Huixin ZHANG ; Jie QIAO ; Yingli LU
Chinese Journal of Endocrinology and Metabolism 2011;27(9):749-751
The fasting and 2 h levels of glucagon, somatostatin ( SS), and C-peptide during 75 g oral glucose tolerance test in 60 patients with type 2 diabetes and 34 normal subjects were determined. Compared with control group, the fasting levels of glucagon and SS and 2 h levels of SS after glucose loading significantly decreased,while the fasting and 2 h levels of C-peptide increased in diabetes group. The 2 h levels of these hormones were significantly higher than the fasting levels in two groups. Compared with control group, the increased folds of glucagon ( 1.40±0.48 vs 1.20±0. 30, P<0. 05 ) and SS( 2.79±2. 17 vs 1.14±0. 22, P<0. 01 ) levels after glucose loading were higher and that of C-peptide level ( 3.58 ±3. 10 vs 8. 33 ± 6. 99, P<0. 01 ) was lower in diabetes group. The levels of fasting glucagon were positively correlated with that of fasting SS in two groups( both P<0. 01 ). These results suggest that disturbance exists in hormones from α and δ cells besides the dysfunction of β cells in patients with type 2 diabetes.
5.Biomechanical study on helical long PHILOS plate fixation for proximal metaphyseal-diaphyseal humeral shaft fractures
Lei WANG ; Yan-hao CHEN ; Chun-xia LU
Journal of Medical Biomechanics 2015;30(5):E463-E467
Objective To compare biomechanical properties of the helical and straight long PHILOS (proximal humerus internal locking system) plates (Synthes Inc., Switzerland), so as to provide some biomechanical evidence for treating proximal metaphyseal-diaphyseal humeral shaft fractures in clinic. Methods Twelve Synbone artificial bones of right humerus (SYNBONE Inc., Switzerland) were divided into two groups. In control group (n=6), the humerus was fixed with the 10 hole long straight PHILOS plate, while in experimental group (n=6), the humerus was fixed with the same long PHILOS plate which was precontoured for moulding (i.e. helical PHILOS plate). After the proximal metaphyseal-diaphyseal humeral shaft fractures were made in all artificial bones, the biomechanical properties of the specimens in two groups under 6 loading modes (i.e., axial tension and compression, torsion in the same and reverse direction, medial-lateral and anterior-posterior three-point bending) were tested en bloc and compared. ResultsCompare with control group, under 100-500 N tensile and compressive loads, the axial displacement at the fractured end in experimental group increased by about 95% and 58%, respectively. Under 0.6-3 N•m torsional moment in reversed direction, the tensional angle in experimental group was obviously smaller than that in control group, with a decrease of 55%-64%. Under medial-lateral bending moment of 1.5 and 3 N•m, no significant difference was found in deflection of the experiment and control group, while under medial-lateral bending moment of 4.5, 6 and 7.5 N•m, the deflection in experimental group decreased by 20%-30% as compared to control group. Under 0.6-3 N•m torsional moment in the same direction and 1.5-7.5 N•m anterior-posterior bending moment, both the torsional angle and the deflection in experimental group were larger than those in control group, with a significant difference (P<0.05). Compared with control group, the tensile stiffness and compressive stiffness decreased by 49% and 36%, the torsional stiffness in the same direction decreased by 19% and that in reversed direction increased by 150%, three-point bending stiffness in medial lateral direction increased by 18% and that in anterior posterior direction decreased by 70% in experimental group, all with a significant difference (P<0.05). ConclusionsCompared with the long straight PHILO plate, the long helical PHILOS plate has better biomechanical properties, which can meet the clinical need of proximal metaphyseal-diaphyseal humeral shaft fracture fixation and postoperative rehabilitation. This surgical technique is expected to be widely applied in clinic, especially with the advantage of minimal invasive surgery.
6.Screening report on endemic arsenism and high content of arsenic in Xiantao City,Hubei Province
Shu-jun, ZHAO ; Gan-lu, LIU ; Bao-xia, YANG ; Zhong-jun, LUO ; Ye-xin, LU ; Xiang-hong, LI ; Fang, WANG ; Shou-jiao, TAN
Chinese Journal of Endemiology 2009;28(1):71-74
Objective To investigate the basic distribution of endemic areas in the type of drinking water arsenism and in Xiantao City,Hubei Province,and to offer a scientific basis for control and prevention.Methods According to"the Chinese Scheme of Implementing Surveillance of Distribution of Endemic Arsenism",considering with the special geography feature of Xiantao,both sampling and overall survey were used in 7 towns chosen.The water arsenic content was determined by half quantitative fast reagent-box method.We began to search for clues and patients according to the endemic areas and the families with high arsenic wells.Results High endemic arsenic water sources were distributed in 7 the towns(districts or farms).In 81 villages of Xiantao City,35 villages had drinking water arsenic content exceeding 0.05 mg/L,accounting for 43.21%(35/81).In 4020 screened wells,269 had arsenic content higher than the national standard,the detective rate of high arsenic wells(more than 0.05 mg/L)was 6.69%(269/4020),with the highest rate in Shahu Seed Plant being 13.56%(115/848).The population exposed to high arsenic was 1091,in a rate of 5.75%(1091/18 975),in which 281 children were exposed in a rate of 5.82%(281/4826).In Shahu Seed Plant,467 people including 129 children were exposed to high arsenic, accounting for 13.26%(467/3522)and 12.91%(129/999),respectively.Conclusions High arsenic sources widely exist in Xiantao City,especially in Shahu Seed Plant,where arsenic content,the exposed rate of population and children are high.Therefore,prevention and control should be carried out in the southeast as soon as possible,as well as in other places where situation is less serious.
7.Expression of COX-2 mRNA in rat aortic endothelial cells in high glucose environment
Yingli LU ; Tingting YE ; Hua YANG ; Yi CHEN ; Jiao YU ; Lijuan ZHAO ; Fangzhen XIA ; Ningjian WANG ; Chao SHI ; Jie QIAO ; Lin YE ; Dongping LIN ; Wanling WU
Chinese Journal of Endocrinology and Metabolism 2009;25(4):382-383
ed in vitro and in vivo,which was inhibited by aspirin treatment.
8.Salidroside protects cultured rat subventricular zone neural stem cells against hypoxia injury by inhibiting Bax, Bcl-2 and caspase-3 expressions.
Cunfang QI ; Junfeng ZHANG ; Xinlin CHEN ; JiansHui ZHANG ; Pengbo YANG ; Qian JIAO ; Pengbo ZHANG ; Hai-Xia LU ; Yong LIU
Journal of Southern Medical University 2013;33(7):962-966
OBJECTIVETo explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury.
METHODSPrimarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells.
RESULTSSalidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (P<0.05). Salidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (P<0.05).
CONCLUSIONSalidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.
Animals ; Caspase 3 ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Flow Cytometry ; Glucosides ; pharmacology ; Neural Stem Cells ; drug effects ; Phenols ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism
9.Histologic subtyping and prognosis of thymoma: a study of 108 cases.
Xia JIAO ; Hong-Lin YIN ; Zhen-Feng LU ; You-Cai ZHAO ; Xiao-Jun ZHOU
Chinese Journal of Pathology 2008;37(7):445-449
OBJECTIVETo study the prognostic and clinical relevance of histologic subtyping of thymoma according to the World Health Organization (WHO) classification.
METHODSThe clinicopathologic features of 108 patients with thymoma removed surgically were retrospectively reviewed. The histologic diagnosis of the tumors was made on the basis of 2004 WHO classification by two experienced pathologists. The correlation between Masaoka tumor stage, WHO histologic subtype, completeness of resection, presence of myasthenia gravis, other clinical parameters (including age, gender and tumor size) and survival was studied.
RESULTSAccording to WHO classification, there were 7 cases (6.5%) of type A thymoma, 19 cases (17.6%) of type AB thymoma, 23 cases (21.3%) of type B1 thymoma, 19 cases (17.6%) of type B2 thymoma, 27 cases (25.0%) of type B3 thymoma and 13 cases (12.0%) of type C thymoma. According to Masaoka tumor staging, 36 cases (33.3%) were in stage I, 34 cases (31.5%) in stage II, 27 cases (25.0%) in stage III and 11 cases (10.2%) in stage IV(a). The association between histologic subtype and Masaoka tumor stage was statistically significant (P = 0.000). The 5-year survival rates of type A, AB, B1, B2 and B3 thymoma cases were 100%, 100%, 93%, 83% and 43%, respectively; while the 10-year survival rates were 100%, 100%, 81%, 70% and 33%, respectively. The median survival time of type C thymoma was 62.5 months. Type B2 and B3 thymoma cases had an intermediate prognostic ranking in comparison with type C thymoma and other groups (P = 0.000). The 5-year survival rates of tumors in stage I, II and III were 100%, 77% and 54%, respectively; while the 10-year survival rates were 100%, 70% and 27%, respectively. The median survival time of patients in stage IV(a) was 14.0 months. Masaoka tumor stage was highly significant in predicting survival of patients (P = 0.000). On multivariate analysis, Masaoka tumor stage was an independent predictive factor for survival (P = 0.027). On the other hand, the WHO subtype (type A to B1 versus type B2 to B3 versus type C) and completeness of resection could predict the tumor-related survival.
CONCLUSIONSThe Masaoka tumor stage is the single most important prognostic factor of thymoma. The WHO histologic subtype and completeness of resection affect mainly the post-operative survival. The classification of thymoma may also reflect the clinical behavior of the tumor. Type A, AB and B1 thymomas belong to the low-risk group, while type B2 and B3 thymomas have an intermediate prognostic ranking. Type C thymoma carries the worst prognosis.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Myasthenia Gravis ; etiology ; Prognosis ; Survival Analysis ; Thymoma ; classification ; complications ; diagnosis ; pathology ; Thymus Neoplasms ; classification ; complications ; diagnosis ; pathology ; World Health Organization
10.Treatment of nonneoplastic epithelial disorders of skin and mucosa of vulva with focused ultrasound
Lu-Xia JIAO ; Li-Na HU ; Zheng-Ai XIONG ; Li-Hua FENG ; Yong ZHANG ; Yun-Bo TIAN ; Wen-Zhi CHEN ; Zhi-Biao WANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(01):-
Objective To explore the clinical efficacy of focused ultrasound for patients with white lesions of the vulva,as well as its safety and feasibility.Methods Clinical data of 941 patients with white lesions of the vulva treated with focused ultrasound from June 2003 to December 2005 were retrospectively reviewed.The mean age of the patients was 40.8 years(18-70 years)and the median course of the disease was 6.2 years(3 months-45 years).Meanwhile,pathological diagnosis was performed in all the patients before treatment,in which 498 cases were squamous hyperplasia,342 cases were lichen sclerosus and 101 cases were lichen sclerosus with squamous hyperplasia.Patients were followed up and therapeutic effects of focused ultrasound was evaluated at 6 and 12 months after the treatment,respectively.The symptoms of pruritus in the vulva and the changes in the color and elasticity of the vulvar lesions were observed.Results Of all the patients,900 were followed up after the treatment,and the ratio of effectiveness was 94.9%.Only 46 patients(5.1%)had no response to the therapy.Of the effective patients,434 cases were completely cured(48.2%),and 420 cases were improved(46.7%).Pruritus of vulva recurred in 101 patients (11.2%)one year after treatment;however,these patients still had a response to the second or third treatment.Conclusions Focused ultrasound therapy is a highly effective instrument in treatment of white lesions of the vulva.It can not only relieve the symptoms of itching,but is also helpful in recovering the color and elasticity of the vulva.