2.Pleiotrophin and neural repair after peripheral nerve injury
Jin LI ; Jianghai CHEN ; Hao KANG
International Journal of Surgery 2008;35(8):552-556
Pleiotrophin, discovered in 1989, was thought as a muhi-funetional growth factor; It plays an important role in tumor occurrence and central neural system. The latest research showed pleiotrophin signal pathway probably takes part in neural repair after peripheral nerve injury, especially in the followed crisis point, such as the protection of spinal cord neuron, the promotion of the speed of neuron axon regeneration, the guidance of neuron axon regeneration, skeleton muscle reirmervation. It potentially makes a key role in the guidance of neural axon regeneration in peripheral neural system and muscle reianervation. Along with the related researches go deep, pleiotrophin gene might become a controllable target for promoting the result of peripheral neural repair and reconstructing the neuromuscular junction.
3.Single nucleotide polymorphisms in cell cycle regulator p21 and p27 genes are associated with susceptibility to epithelial ovarian cancer
Xia JIN ; Shan KANG ; Na WANG ; Yanping XING ; Yan LI
Chinese Journal of Obstetrics and Gynecology 2008;43(3):209-212
Objective To investigate the association of single nucleotide polymorphisms(SNP)in p21and p27 genes with the risk of epithelial ovarian cancer(EOC).Methods Genotypes were analyzed by polymerase chain reaction-restrictive fragment length polymorphism(PCR-RFLP)method in 234 patients with EOC and 284 control women in China.Results (1)The frequencies of the p21 in healthy controls were 34.2%.49.6%and 16.2%,while the distribution of the C and T allele was 59.0%and 41.0%,respectively.The p21 C/C(28.2%),C/T(53.0%),T/T(18.8%)distribution in ovarian cancer patients was not significantly different from that in healthy controls(P>0.05).There was no statistic difference in allele distribution between ovarian cancer patients and healthy controls(P>0.05)either.The stratification analysis by tumor histological type did show that the genotype distribution in four types of ovarian cancer patients was significantly different from that in healthy controls(P=0.02).The C/C genotype was likely to reduce the risk of epithelial endometrial cancer.and the adjusted odds ratio was 0.56(95%CI:0.32-0.98).(2)The genotype frequencies of the p27 in healthy controls were 88.4%,10.9%and 0.7%.while the distribution of the V and G allele was 93.8%and 6.2%.respectively.The V/V(93.6%),V/G(5.1%)and G/G(1.3%)distribution in ovarian cancer patients was significantly different from that in healthy controls(P=0.04).There was no statistic difference in allele distributionbetween ovarian cancer patients and healthy controls(P>0.05).Compared with the V/G and G/G genotypes,the V/V genotype increased the risk of EOC,the adjusted odds ratio was 1.92(95%CI:1.02-3.63).Conclusion The C/C genotype of p21 may reduce the risk of epithelial endometrial cancer,and the genotype of p27 V/V may be a potential risk factor for susceptibility to EOC.
4.Clinical study on repairing huge soft tissue defect in foot with a extensive reverse peroneal artery flap
Jin LI ; Hao KANG ; Yanhua CHEN ; Fabin WANG ; Guangxiang HONG
Chinese Journal of Microsurgery 2009;32(3):184-186,illust 2
Objective To introduce the clinical application of a extensive revense peroneal artery flap in repairing big soft tissue defect in foot, especially in repairing a big defect in distal foot and the exposure of one or more metatarsophalangeal joint. Methods The skin flaps were used in 11 cases to repair the big soft tissue defect in foot, including the distal part of foot. The flap upper boundary may surpass the capitulum fibulae; The lower boundary may reach the back line between external malleolus and internal malleolus; Lat-eral boundary could overrun anterior margin of fibula about 2 cm; Inboard boundary could get to medial mar-gin of gastrocnemius muscle. In attention: the two branch vessels from peroneal vessel in the site of 11-13 cm and of 5-7 cm above the outer anker would be included in the flap. Results Ten of 11 skin flaps survived satisfactory. A small, marginal portion of one flap in one case presented necrosis. Conclusion The reverse peroneal artery flap is easy to elevate and safety, which is an ideal flap for a huge soft tissue defect in foot, especial in the distal part of foot.
5.Establishment and optimization of two-dimensional electrophoresis for myocardial proteomes of mice
Ming LIU ; Huijun WANG ; Xuefeng LI ; He JIN ; Wei KANG
Chinese Journal of Tissue Engineering Research 2006;10(28):174-176,封三
BACKGROUND: Two-dimensional electrophoresis (2-DE) is precise and effective for the isolation and analysis of complex protein mixtures, has become one of valuable key techniques for proteome. Mice models were as research subjects to model human disease status, one of important methods for cardiovascular disease. The establishment and optimization of 2-DE for proteomes of mice myocardium will provide basis for heart disease.OBJECTIVE: To establish and optimize a stable technique of 2-DE for proteomic analysis of mice myocardium.DESIGN: Mice were used as research subjects, observational comparative study.SETTING: Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University.PARTICIPANTS: The experiment was performed at the Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University between February and September 2004. Totally 12 male BABL/c mice of 4-5 weeks without special-pathogen free with the body mass of 12-15 g were selected, which was provided by the Experimental Animal Center of Southern Medical University.METHODS: The mice were put to death by dislocation of cervical vertebra under drugged state to obtain heart. Various protein extraction method,loading sampling buffer, strip range of IPG, sampling volume of protein,staining method and so on were compared, analyzed, scanned and digilized,and then performing image analysis. Each link in the study was established and optimized.MAIN OUTCOME MEASURES: Resolution of protein and reproducibility of the trial.RESULTS: A method that was optimal for cardiac tissue can isolate about a total of 910 protein spots on pH 4-7 gel strips. 200 μg was fit for silver stain while 1000 μg was for Coomassie stains. It was found that in the myocardium of normal mice about (920±30) protein spots were obtained in 17 em pH 3-10 L gel strip, while around (880±30) were gained in the pH 4-7 L gel strip. The mean matching rate achieves to 86. 9%.CONCLUSION: The findings of the trial showed that 2-DE technique can be effectively applied for proteomics of myocardium of mice to regulate and optimize, and relatively stable separation method of myocardial protein was established.
6.Establishment of leukocyte-associated immunoglobulin like-receptor 2 (CD306) eukaryotic expression vectors and purification and identification of fusion protein
Chunyan WANG ; Zhenhua KANG ; Xin XIE ; Yan LI ; Boquan JIN
Chinese Journal of Tissue Engineering Research 2009;13(50):9928-9932
BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.
7.The effects of basic characteristics of patients on non-motor symptoms and the relationship between motor symptoms and non-motor symptoms in patients with Parkinson's disease
Jun LI ; Miao JIN ; Xiaoyun ZHENG ; Kang WANG
Chinese Journal of Geriatrics 2017;36(6):618-621
Objective To investigate whether sex,age-of-onset,education and asymmetry affect non-motor symptoms (NMS)in Parkinson disease,and to analyze the relationship between motor symptoms and NMS in Parkinson's disease.Methods The detailed clinical information of 157 patients with idiopathic Parkinson's disease(PD)was recorded and followed up.The data was calculated with SPSS statistic software.Sex had an impact on sleeping disorder (P< 0.05).Sleep disorder in female was more prominent.Cognitive disorder was affected by age-of-onset and education duration.The pain and sensory disorder were affected by age-of-onset (P < 0.05).Differences in the effects of asymmetry on NMS were not statistically significant (P> 0.05).The higher the scores of postural instability and gait difficulty (PIGD)were,the lower the Mini-Mental State Examination (MMSE)score was(β=-1.837,P =0.004).The higher the tremor score was,the higher the Hamilton Depression Scale(HAMD) score was(β=3.460,P =0.001).The higher the rigidity akinesia score was,the higher the autonomic dysfunction score was(β=0.104,P=0.006).Conclusions The non-motor symptoms of Parkinson's disease are affected by the age,sex and cultural level.Symmetryor-not does not affect Parkinson's disease.Non-motor symptoms and motor symptoms are closely linked.The encountered posture and gait abnormalities of patients should trigger physician alerts to focus on patients' cognitive function.The encountered tremor of patients should trigger physician alerts to focus on their emotional changes.Facing to the patient with rigidity and less moving,physician should alert to the occurrence of autonomic symptoms.
8.Mechanism of apoptosis of nasopharyngeal carcinoma cells induced by polysaccharides extracts from Hedyotic diffusa.
JING YAN ; KANG MIN ; LIU JIN ; LI JINGYU ; TANG ANZHOU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(7):641-644
OBJECTIVE:
To explore the proliferation inhibition and apoptosis of polysaccharides extracts from polysaccharides extracts from Hedyotic diffusa (PEHD) on Human Nasopharyngeal Carcinoma (NPC)cell line CNE2 cells in vitro.
METHOD:
CNE2 cells treated with various concentrations of PEHD were detected by MTT assay at 24 h, 48 h, and 72 h. The apoptotic cells were analyzed by flow cytometry with Annexin V/PI staining. The expression levels of Bax, Bcl-2 and caspase-3 protein were examined by Western blotting method.
RESULT:
The growth of CNE2 cells were suppressed after treatment with PEHD (P < 0.05), MTT assay showed that the highest cell inhibition rate reached to 76.5%, the inhibition in the doses from 2 to 6 mg/ml showed dose-and-time-dependent. The percent of apoptosis in 4 and 6 mg/ml PEHD treatment groups for 48 h were 31.32%, 46.28%, respectively, and significantly higher than that in control groups, 4.86% (P < 0.01). After the cells being treated with PEHD for 48 h, the expression of Bax and caspase-3 protein increased, and the expression of Bcl-2 protein decreased gradually.
CONCLUSION
PEHD could inhibited the growth of CNE2 cells and was dose-and-time-dependent, the mechanism may involve induction of cell apoptosis, which was associated with the activation of Bax and caspase-3 protein and the down-regulation of Bcl-2 protein expression.
Apoptosis
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Carcinoma
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Caspase 3
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metabolism
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Cell Line, Tumor
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drug effects
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Cell Proliferation
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Down-Regulation
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Hedyotis
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chemistry
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Humans
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Nasopharyngeal Carcinoma
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Nasopharyngeal Neoplasms
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pathology
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Plant Extracts
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pharmacology
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Polysaccharides
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pharmacology
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Proto-Oncogene Proteins c-bcl-2
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metabolism
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bcl-2-Associated X Protein
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metabolism
9.Preoperative full reset combined with minimally invasive treatment of extreme distal pilon fractures
Jin KANG ; Yongle LI ; Xiaowei LIU ; Tiejun LI ; Ming ZHAO ; Chao LI ; Xiandong FAN
Chinese Journal of Trauma 2016;32(10):915-920
Objective To investigate the clinical efficacy of preoperative full reset combined with minimally invasive treatment of extreme distal pilon fractures.Methods A retrospective analysis was made on 34 patients (35 ankles) with tibial fractures extremely close to the distal articular surface treated surgically between January 2011 and January 2015.There were 21 mnales and 13 females,aged 20-71 years (mean,36.2 years).Injury resulted from traffic accidents in 32 patients and high falls in two.Using the AO/OTA fracture classification system,type 43-B3 was noted in three patients,43-C1 in five patients,43-C2 in 18 patients and 43-C3 in eight patients.Calcaneal traction combined with manipulative reduction was used to correct fracture displacement preoperatively.All fractures were stabilized by minimally invasive percutaneous plate osteosynthesis (MIPPO) through single or combined medial,anteromedial and anterolateral approaches while minimizing damage to bone attachment and continuity of soft tissue,after soft tissue swelling subsided.For the patients with articular surface collapsing with severe comminution,a series of procedures were done under direct vision including using the talus articular surface as a mold,stable fixation with fine Kirschner (1-1.5 mm) and thin screws (2.1-2.7 mm series) and impaction bone grafting below subchondral bone.Thereafter,distal tibia anatomical short multi-directional locking plate fixation,distal nail support and early ankle joint functional exercise were done.Burwell-Charnley radiological evaluation system was used for radiological assessment,and TeenyWiss scoring system for ankle clinical symptoms and function.Postoperative complications were recorded.Results Follow-up lasted for 11-38 months (mean,16.6 months).No infection,wound disunion,or plate exposure occurred.Burwell-Charnley radiological evaluation system showed anatomic reduction in 32 patients,unsatisfactory reduction in one,and poor reduction in one.According to the Teeny-Wiss scoring system,the results were excellent in 31 patients,good in two and poor in one,with the excellentgood rate of 97%.Three patients suffered traumatic arthritis after operation and alleviated after oral administration of painkiller.Conclusion With use of full reset combined with manipulative reduction to correct fracture displacement,minimally invasive locking plate,distal row of nails,impaction bone grafting and limited fixation,the patients with extremely distal tibial pilon fractures achieve satisfactory reduction,stable fixation,and early functional exercise.
10.Expression of Heat Shock Protein-70 in Paraquat-induced Pulmonary Inflammation Response
Jingdong LI ; Kang JIN ; Yunhe YANG ; Shan YU ; Junping SUN ; Tian JIN
Journal of China Medical University 2010;(3):212-214,227
Objective To investigate the expression of heat shock protein-70(HSP70)in acute lung injury induced by paraquat poisoning.Methods A total of 72 Wistar rats were randomly divided into 3 groups(n=24 each group)and intraperitoneally injected with normal saline in group A,20 mg/kg paraquat in group B,and 40 mg/kg paraquat in group C.The lungs of rats were excised 1,3,6,12,and 24 hours after the injection,respectively.The expression of HSP70 in the lung tissue was detected by immunohistochemistry and Western blot.Results Paraquat induced acute lung injury in groups B and C.In group B,the expression of HSP70 increased at 2 hours,reached the peak at 6 hours,decreased at 12 hours,and returned to the baseline level at 24 hours.In group C,the expression of HSP70 increased at 1 hour,reached the peak at 3 hours,and returned to the baseline level at 24 hours.Conclusion Paraquat-induced acute lung injury may contribute to the enhanced expression of HSP70 in the bronchus,bronchiole,and alveolar epithelial cells,which indicates that HSP70 has a protective effect on lung injury.