1.In vitro morphological observation on the effect of exogenous collagen on the construction of tissue-engineered tendon
Yue FANG ; Zhou XIANG ; Zhiming YANG
Chinese Journal of Tissue Engineering Research 2006;10(29):169-172
BACKGROUND: Collagen type Ⅰ possesses low antigenicity and provides special functions of scaffold structure for cell migration, proliferation and secretion.OBJECTIVE: To observe the morphological change after combination of transformed human embryonic tendon cells and artificial materials with collagen type Ⅰ, so as to provide the experimental basis for constructing tissue-engineered muscle tendon in vitro.DESIGN: Randomized and controlled observation.SETTING: Department of Orthopaedics, West China Hospital, Sichuan University.MATERIALS: This experiment was carried out at the Department of Transplantation Immunology, West China University of Medical Sciences from July to September 1998. Human hair, carbon fiber and polyglycolic acid were combined with collagen and then set corresponding groups respectively. Simply hair, carbon fiber and polyglycolic acid control groups were set respectively. Artificial materials: Healthy adult hair (30 cm in length, 80 g in weight), carbon fiber knitted strip (Lanzhou carbon factory )and polyglyolic acid fasciculi (Johnson , American) were used in this experiment.METHODS: ①In the group of human hair combined with collagen,nine healthy adult hairs were chosen . Three hairs as one bundle were knitted into tendon with 0.8 cm in length and 0.4 cm in diameter and a knot was tied respectively at the two ends. ②In the carbon fiber combined with collagen group, the purchased carbon fiber knitted strip was separated and made into artificial materials with the same diameter and length as those of human hair tendon. Two ends of the artificial materials were packed with 5/0 silk thread. ③In the polyglycolic acid combined with collagen group, three polyglycolic acid fasciculi were weaved into bundle with the same diameter and length as those of human tendon,and two ends were packed with 5/0 silk thread. ④The process of knitting scaffold materials was the same in control groups as in the corresponding experimental groups. ⑤ Five pieces were prepared in each scaffold material. After sterilized by 60Co, the scaffold materials were put in collagen type Ⅰ solution for 0.5 hour, then taken out and dried at roomtemperature for use in the experimental groups. ⑥transformed human embryonic tendon cell, density for 3×106/mm3, was used for combined culture in each group, and cell number and morphology were observed at different time points after culture under an inverted microscope and scanning electron microscope.MAIN OUTCOME MEASURES: ① Observation of cell growth and adherence under an inverted microscope ② Observation of cell morphology and material adherence under scanning electron microscope RESULTS: ① Observation of cell growth and adherenceunder an inverted microscope :In each experimental group, at two hours after culture, cells gathered around the materials, and some cells adhered to materials presented round; At three days after culture, the number of cells adhered to materials began to increase, morphology changed from round to oval, and the co-existence of round and oval cells was seen;While in each control group, at two and three hours after culture, few cells adhered to the three materials. On day five after culture, in each experimental group, cell morphology turned into fusiform, the number of cells increased further and a great many fusiform cells grew among carbon fiber materials; while in the control group, most cells presented round and ellipse, and the number of cells was less as compared with the corresponding experimental group. ② Cell morphology and material adherence under scanning electron microscope: On day 5 after in vitro culture, most tendon cells on the surface of materials presented spindle-shape in each experimental group. Apophysis stretched out from the tendon cells, grew along longitudinal axon of the materials. Silk and piece-like substance appeared among the apophysis and weaved each other. While there were fewer cells adhered in control group than in corresponding experimental group, cells also presented fusiform or round, but no silk or piece-like substance was found, also no apophysis appeared.CONCLUSION: Human hair, carbon fiber and polyglycolic acid combined with collagen type Ⅰ will facilitate cell adherence. Cells increase around the materials and co-existence of round and oval cells could be seen, and cells turned into fusiform, suggesting that exogenous collagen facilitate the cells to adhere onto the materials and proliferate.
2.Expression and Clinical Significance of p27~(KIP1) and p57~(KIP2) in Thyroid Carcinoma
Jierong TANG ; Zhiming WANG ; Xiang CHENG
Journal of Chinese Physician 2000;0(11):-
Objective To explore the relationship between the expression level of p27 KIP1 and p57 KIP2 and the development and prognosis of thyroid carcinoma. Methods p27 KIP1 and p57 KIP2 expression in 6 nodular goitres, 10 thyroid adenomas and 35 thyroid carcinoma were detected by immunohistochemical method, and the relationship between p27 KIP1 and p57 KIP2 expression and clinicopathological characteristics were analyzed. Results The positive rate of nuclear expression of p27 KIP1 and p57 KIP2 in nodular goitres and thyroid adenomas was obviously higher than that in thyroid carcinomas, in papillary carcinoma and follicular carcinoma was significantly higher than that in medullary carcinoma and undifferentiated carcinoma, and in the thyroid carcinomas of stages I and II was also significantly higher that in the thyroid carcinomas of stages III and IV. The frequency of cytoplasmic expression of both p27 KIP1 and p57 KIP2 in thyroid carcinoma with lymph node metastasis was obviously higher than that in ones without lymph node metastasis. Conclusion The nuclear expression of both p27 KIP1 and p57 KIP2 in thyroid carcinoma were significantly associated with clinical stage, lymph node metastasis and local invasion. The low nuclear expression of p27 KIP1 and p57 KIP2 may indicate the poor prognosis of the patients, and the cytoplasmic expression may indicate the lymph node metastasis.
3.Delayed onset muscle soreness and muscle satellite cells:repair of skeletal muscle injury
Xiang ZHANG ; Zhiming CHAI ; Li ZHAO
Chinese Journal of Tissue Engineering Research 2015;(37):6031-6036
BACKGROUND:Skeletal muscle fiber damage that is induced by prolonged or high-intensity exercise directly cause muscle injury. OBJECTIVE:To propose the existence of a relationship between delayed onset muscle soreness and muscle satel ite cel s after skeletal muscle injury from the objective reality. METHODS:A retrieval of CNKI and PubMed databases was done for relevant literature published from February 1961 to February 2015 using the keywords of“delayed onset muscle soreness;skeletal muscle injury;satel ite cel s, skeletal muscle;myogenic regulatory factors”in Chinese and English, respectively. Final y, 59 articles were included to explore the mechanism of skeletal muscle injury accompanied by delayed onset muscle pain. RESULTS AND CONCLUSION:Skeletal muscle micro-injury mainly refers to ultrastructural destroy and metabolic disorders, involving muscle cel membrane, cytoskeleton, sarcomere and mitochondria, thereby leading to skeletal muscle contraction dysfunction that is often accompanied with delayed onset muscle soreness. Eccentric exercise can cause the severest microdamage to the skeletal muscle fibers. The occurrence of delayed onset muscle soreness can stimulate the secretion of growth factors from skeletal muscle satel ite cel s to a certain extent, and the time series of delayed onset muscle soreness triggered by sustained high-intensity eccentric exercise has a certain correlation with the time series of the proliferation of skeletal muscle satel ite cel s.
4.Comparison of complete-block radical gastrectomy and traditional radical gastrectomy
Fenghua GUO ; Fen LUO ; Xiang MAO ; Jun WANG ; Zhiming WANG
Fudan University Journal of Medical Sciences 2010;37(1):103-105,123
Objective To introduce the techniques and specification of complete-block radical gastrectomy, and to compare with traditional radical gastrectomy in the operative time, surgical costs and postoperative complications. Methods Thirty gastric cancer patients meet the criteria were randomly divided into 2 groups. Group A: complete-block radical gastrectomy;group B: radical gastrectomy. Analysis and comparison were made on tumor site, time consuming of block, the proportion of gastrointestinal reconstruction, operative time, cost of block, pathological TNM stage, length of stay, postoperative complications and postoperative chemotherapy. Results Group A has longer operative time than group B (P<0.05). However, there was no significant statistical difference in the other indicators between these 2 groups. Conclusions Complete-block technology can separate cancer more completely during operation with the advantage of simplicity, low-cost, little effect on the subsequent operation. The incidence of postoperative complications and length of stay with the traditional surgery there was no significant difference.
5.Immunological characteristics of Mycobacterium tuberculosis antigen Rv2628.
Yuelan YIN ; Yunfei GAO ; Dan ZHAO ; Kai LIAN ; Xiang CHEN ; Zhengzhong XU ; Zhiming PAN ; Xin'an JIAO
Chinese Journal of Biotechnology 2014;30(2):255-264
Antigen Rv2628 of Mycobacterium tuberculosis is associated with latent tuberculosis infection. In this study, Rv2628 was prokaryotic expressed and purified, its immunological characteristics was evaluated with macrophage cell line RAW264.7 and BALB/c mice. The results show that Rv2628 was mainly expressed in form of inclusion body confirmed by SDS-PAGE, and could react with rabbit anti-H37Rv polyclonal antibody detected by Western blotting assay, indicating that the protein had an effective immunoreactivity. The interactions between Rv2628 and macrophage cell line RAW264.7 confirmed that it could effectively induce cells to produce pro-inflammatory cytokines, the relative expression level of IL-6 mRNA was higher than the control group in 1-12 h. BALB/c mice were subcutaneously immunized with Rv2628 protein, the production of IFN-gamma and IL-4 in the spleen cells was determined by Sandwich ELISA, in the Rv2628 immunized group, the level of IFN-gamma was significantly higher than that of IL-4 (P < 0.000 1). It indicated the protein induced Th1-tendency immune responses. At the same time, Rv2628(11-30) peptide used as coating antigen, the murine serum antibody titer detected by indirect-ELISA was 1:1 600, which demonstrated that Rv2628 could also induce humoral immune responses. In summary, Rv2628 could induce specific pro-inflammatory cytokines, affectively induce strongly Th1-tendency immune response and humoral response, it could be a potential target for developing subunit vaccine against TB. In addition, it laid foundation for probing the cross-talk between M. tb and host.
Animals
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Antibodies, Bacterial
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blood
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Antigens, Bacterial
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immunology
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Cell Line
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Electrophoresis, Polyacrylamide Gel
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Interferon-gamma
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immunology
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Interleukin-4
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immunology
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Interleukin-6
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immunology
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Macrophages
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immunology
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Mice
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Mice, Inbred BALB C
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Mycobacterium tuberculosis
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Th1 Cells
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immunology
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Tuberculosis
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immunology
7.Analysis of clinicopathological characteristics and prognosis for triple negative breast cancer
Xiang LUO ; Yanxia SHI ; Zhiming LI ; Dongsheng ZHANG ; Zhengyan SU ; Wenqi JIANG
China Oncology 2009;19(7):517-522
Background and purpose: Triple negative breast cancer (TNBC) is a high risk breast cancer characterized by the negative expression of estrogen receptor(ER), progesterone receptor (PR) and Her-2 that have no specific therapy. This study was to analyze clinical pathological characteristics, survival, and prognostic factors of patients with TNBC. Methods: Clinical and pathological as well as follow-up data of TNBC, treated at the Cancer Centre of Sun Yat-sen University from Jan. 2000 to Dec. 2003, were collected and analyzed. Results: A total number of 128 women were identified as having triple negative breast cancer. The median age of these patients was 46 years, and 60.9% of them had stage Ⅰ or Ⅱ disease. The majority of pathological types were invasive ductal carcinomas, and 78.1% of tumors were staged T1 or T2. And 48.4% of these patients were involved in lymph node. Event-free survival, local replase-free survival, distant metastasis-free survival and overall survival at five years were 71.1%, 84.3%, 75.8% and 83.6% respectively. Though lymph node metastasis, tumor masses, stage and lymph-vascular invasion were all found to be related to overall survival, however, only lymph node metastasis and tumor masses affected the overall survival as revealed by the Cox proportional hazard model analysis. Conclusion: Triple negative breast cancer has distinct clinical and pathological characteristics. The patients are usually young, with large masses, lymph node metastasis, family history of breast cancer and poor prognosis; lymph node metastasis and tumor mass are important prognostic factors.
8.Prediction of CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains in China
Zhiming FANG ; Hui XING ; Zhefeng MENG ; Kunxue HONG ; Lingjie LIAO ; Xiang HE ; Jianxin Lü ; Yiming SHAO
Chinese Journal of Microbiology and Immunology 2010;30(2):135-139
Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.
9.The analysis of quinolone resistance of the avian Escherichia coli and Salmonella isolates from 1993 to 2008
Weijuan PAN ; Xiang CHEN ; Xiaoquan WANG ; Qiuxia CONG ; Zhiming PAN ; Song GAO ; Xinan JIAO
Chinese Journal of Zoonoses 2009;(7):630-635
Quinolones are broad-spectrum antibacterial agents used in human and veterinary medicine, and their extensive use have been associated with a rise of the quinolone resistance. In the present study, the quinolone resistance of avian E.coli and Salmonella isolates was evaluated and compared, in which 344 avian E.coli and 224 Salmonella isolates from 1990s were serogrouped with antisera and thc antimicrobial susceptibility test to 10 quinolones was carried out by using the Kirby-Bauer method recommended by Clinical and Laboratory Standards Institute (CLSI). It was demonstrated that the 344 isolates of avian E.coli distributed in 27 serogroups and 68.90% (237/344) of the isolates belonged to four O-serogroups: i.e. O1, O2, O18, O78, and the 224 isolates of avian Salmonella were all determined to be Salmonella pullorum. The drug-resistance rate of avian E. coli isolates to nalicixic acid from 1993-1999 was more than 60%(64.43%,131/181), whereas those of isolates to 9 antibiotics from 2000-2008 had a drug-resistance rates of more than 60%, namely,nalicixic acid(92.02%), fleroxacin(79.75%), pipemidic acid(79.14%), enrofloxacin(78.53%), enoxacin(76.07%), lomenfloxacin(74.85%), ciprofloxacin(69.33%), norfloxacin(63.80%) and ofloxacin(61.35%). For the 4 O-serogroups of the avian E.coli isolates, the drug-resistance rates of more than 50% to antimicrobials were as follows: O78 isolates to 7 antimicrobials;O18 isolates to 5 antimicrobials, and O1 and O2 isolates just to 3 antimicrobials. The quinolone resistance of Salmonella isolates was much lower than E.coli, in which 101 salmonella isolates from 1993-1999 were all susceptible to quinolones. Nalicixic acid resistance of salmonella isolate firstly appeared in 2000, and the drug-resistance rate of salmonella isolates from 2000-2008 was found to be more than 60% for nalicixic acid(83.74%), but those to other quinolones were comparatively lower. These results indicated that the quinolone resistance of avian E.coli and salmonella were increasing in the past two decads because of the over-use of antibiotics.
10.The changes of VEGF level in patients with hepatocellular carcinoma after argon-helium cryoablation combined with transcatheter arterial chemoembolization treatment
Ding LI ; Hua XIANG ; Zhiming ZHANG ; Jueshi LIU ; Zhiyong FANG ; Lin LONG
Journal of Interventional Radiology 2015;(5):400-403
Objective To investigate the therapeutic effect of argon-helium cryoablation combined with transcatheter arterial chemoembolization (TACE) for primary hepatocellular carcinoma (HCC) and its influence on vascular endothelial growth factor (VEGF) level, and to compare it with simple TACE treatment. Methods During the period from Aug. 2013 to Aug. 2014 a total of 50 patients with primary HCC were admitted to Hunan Provincial People’s Hospital. The patients were randomly and equally divided into group A (TACE group, n=25) and group B (argon-helium cryoablation+TACE group, n=25). VEGF levels in the peripheral blood were determined before and after the treatment, and the results were statistically compared between the two groups. Results After the treatment the VEGF level in group A was significantly increased when compared with preoperative VEGF level (P<0.05). In group B, the VEGF level was decreased after the treatment, which was statistically significant lower than the preoperative VEGF level (P<0.05). The differences in VEGF level between the two groups were statistically significant (P<0.05). Conclusion The statistic analysis of VEGF levels indicates that argon-helium cryoablation combined with TACE is probably superior to simple TACE in the inhibition of tumor angiogenesis.