1.Target gene study in gene transfer technology of cartilage tissue engineering
Liang YANG ; Jianxin LI ; Wenliang WANG
Chinese Journal of Tissue Engineering Research 2010;14(7):1278-1281
OBJECTIVE: To elaborate cartilage tissue engineering in the gene transfer technology and its application, in addition, to make a prospects for its further application.METHODS: The database of Science Direct database (2003-01/2009-04) and CNKI (2003-01/2009-04) were retrieved with key words of "cartilage tissue engineering, gene transfer". The literature was limited to English and Chinese languages. Literatures concerning cartilage tissue engineering in the gene transfer technology were selected, including clinical research and basic research. Other unrelated papers were excluded. Chondrocyte differentiation and gene expression were observed.RESULTS: A total of 90 literatures were searched by computer, according to inclusive and exclusive criteria, the papers regarding cartilage tissue engineering in the gene transfection and gene types and options were analyzed. Gene transfer technology in the field of cartilage tissue engineering has broad application prospects. How to select genes associated with cartilage repair as the transfacted gene need urgent solution. Currently, the used target gene can be divided into following categories, including stimulated cartilage cell proliferation and differentiation, matrix formation, inhibit chondrocyta hypertrophy and osteoblast differentiation, anti-inflammatory response, inhibit senescence and inhibit apoptosis.CONCLUSION: It has a special significance to select the appropriate target genes, and to use a safe gene transfer method to repair cartilage. The clinical application of gene transfer technology is depended on the construction of safe and effective carriers,target genes, as well as transfection systems.
2.Application of adipose tissue-derived stem cells in tissue engineering
Jianxin LI ; Liang YANG ; Wenliang WANG
Chinese Journal of Tissue Engineering Research 2010;14(7):1274-1277
BACKGROUND: Adipose-derived mesenchymal stem cells have similar morphological and biological characteristics to bone marrow-derived mesenchymal stem cells, which can be used as sources of seed cells for tissue engineering.OBJECTIVE: To understand the biological characteristics of adipose-derived mesenchymal stem cells, and to explore its clinical application and prospects in tissue engineering field.METHODS: The databases of PubMed and CNKI were searched with key words of "adipose tissue-derived stem cell, tissue engineering, and stem cells". Literature search was limited to English and Chinese languages. The ossification potential of adipose-derived mesenchymal stern calls and the outcomes combined adipose-derived mesenchymal stem cells with gene transfection to treat diseases were served as evaluative indicators. The in vitro study of comparison between bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stern cells in ossification was included, and irrelative or repetitive papers were excluded.RESULTS AND CONCLUSION: Adipose tissue-derived stem cells can be obtained in large numbers from adipose tissue, and stably proliferate and differentiated in vitro, which possess the similar characteristics to bone marrow-derived menchymal stem call in morphology and biology. Under certain induction, the adipose tissue-derived stem cells can directional differentiated into all three germ layers of cells. Combined adipose tissue-derived stem cells with tissue engineering scaffold could be used to repair bone and articular cartilage defects, but the quality and the surrounding cartilage connecting cartilage, bio-mechanical strength, and future normal cartilage degradation have a certain gap to normal cartilage. With the understanding of adult mesenchymal stem cell research, it found that the self-amplification and differentiation potential of mesenchymal stem cells can effective disused the import of "exogenous gene", thus, it is easy to in vitro genetic modification. Therefore, the adipose-derived stem calls can be combined with genetic engineering, and applied to gene therapy. However, in the present research, the remaining potential carcinogenicity in the gene vector and the negative impact after transfection has not been clarified.
3.Ethanol-induced apoptosis of hepatoma cell line HCC-9204 and its relationship to Bax and Bcl-2 proteins
Lianjun YANG ; Wenliang WANG ; Xiaohui SI
Journal of Cellular and Molecular Immunology 2001;17(4):315-317,358
Aim To explore the ethanol-induced apoptosis effect on hepatocellular carcinoma(HCC) cells and its relationship to the expression of apoptosis associated genes, bax and bcl-2. Methods The cytotoxic effect of 20~ 100 mL/ L ethanol on HCC cell line HCC-9204 was tested by thiazolyl-blue (MTT) assay. Then apoptosis of HCC-9204 cells was induced with 60 mL/ L of ethanol for 6 h. The morphological change, DNA breakage and the change of DNA content of different cell cycles of the apoptotic cells were detected by May-Grunwald Giemsa(MGG) staining, and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and flow cytometer respectively. The changes of expression level of Bax and Bcl-2 proteins were detected by immunocytochemical staining and image analysis. Results The higher the concentration of ethanol was, the stronger the cytotoxic effect on HCC-9204 cells was. 60 mL/ L of ethanol could lead to obviously morphological apoptotic changes of HCC-9204 cells, and majority of the cells were TUNEL positive by TUNEL labeling assay. Typical apoptotic sub G1 peak was observed by flow cytometer. The level of Bax protein expression increased significantly after induced with 60 mL/ L of ethanol for 6 h, no expression of Bcl 2 were found before and after induced with ethanol. Conclusion Low dose of ethanol can induce apoptosis of HCC-9204 cells obviously, and occurance of the apoptosis is related to the increase of the level of Bax protein expression.
4.Effect of surgical trauma on Toll-like receptor 4 expression in hippocampus of aged mice
Wenliang LU ; Xianhui YANG ; Tieli DONG
Chinese Journal of Anesthesiology 2015;(12):1439-1442
Objective To investigate the effects of surgical trauma on Toll?like receptor 4 ( TLR4) expression in the hippocampus of aged mice. Methods Ninety male Kunming mice, aged 16-18 months, weighing 30-40 g, were randomly divided into 3 groups ( n=30 each) using a random number table:control group ( group C), anesthesia group ( group A), and partial hepatectomy group ( group PH). Normal saline 0.1 ml∕10 g was injected intraperitoneally in group C. In group A, fentanyl 0.2 mg∕kg and droperidol 5 mg∕kg were injected intraperitoneally. In group PH, fentanyl 0. 2 mg∕kg and droperidol 5 mg∕kg were injected intraperitoneally, and the mice underwent partial hepatectomy. Cognitive function was assessed using Morris water maze test at 1, 3, and 7 days after anesthesia or surgery. After the end of the test, the hippocampus was immediately harvested for determination of the TLR4, tumor necrosis factor?alpha ( TNF?α) and interleukin?1 beta ( IL?1β) protein and mRNA expression by Western blot and real?time reverse transcriptase?polymerase chain reaction, respectively. Results Compared with group C, no significant changes were found in group PH in the escape latency, percentage of swimming distance in the target quadrant, and TLR4, TNF?α and IL?1β protein and mRNA expression at each time point after anesthesia in group A (P>0.05), and the escape latency was significantly prolonged, the percentage of swimming distance in the target quadrant was decreased, and the expression of TLR4, TNF?α and IL?1βprotein and mRNA was up?regulated at 1 and 3 days after surgery in group PH ( P<0. 05 or 0. 01 ) . Compared with group A, the escape latency was significantly prolonged, the percentage of swimming distance in the target quadrant was decreased, and the expression of TLR4, TNF?αand IL?1βprotein and mRNA was up?regulated at 1 and 3 days after surgery in group PH (P<0.05 or 0.01). Conclusion Surgical trauma can up?regulate the expression of TLR4 in the hippocampus of aged mice, which may be involved in the mechanism of surgical trauma?induced postoperative cognitive dysfunction.
5.Short-term follow-up effect of total hip replacement in fresh and old femoral neck fractures of middle-aged and old patients
Liu YANG ; Lin GUO ; Wenliang WANG
Chinese Journal of Trauma 2003;0(11):-
Objective To compare the short-term follow-up cu ra tive effect of primary total hip replacement (THR) in treating fresh and old fem oral neck fractures. Methods Double follow-ups with an in terval of 13 months were performed in patients with femoral neck fractures under going THR in our department from January 1997 to October 1999. Results After a follow-up for 37-62 months (mean 44 months), we obtained f ull data of 47 patients [41-77 years old, (61.8?5.22) years old] with preoperative Harris score of 28.40?10.14 and postoperative score of 85.20?7. 91. The Harris score of patients with fresh femoral neck fractures (FFNF) incr eased from 21.60?8.73 preoperatively to 82.50?9.81 postoperatively〔at age of (63.50?5.53) years〕 and that of patients with old femoral neck fracture (O FNF) from 37.70?13.50 to 90.63?5.79, [at age of (58.90?7.81) years]. Of 47 patients, only one case had dislocation once within six weeks postoperative ly during function exercise and recovered after manual reduction. Another one wi th excellent function evaluation was given again internal fixation three years a fter THR because of femoral shaft fracture and obtained satisfactory recovery. N o complications were found in other patients. The mean Harris score in patients with OFNF was significantly higher than those with FFNF ( P
6.Early rehabilitation on 180 patients with severe brain injury
Hao ZHOU ; Zhi YANG ; Wenliang ZHAO
Chinese Journal of Rehabilitation Theory and Practice 2003;9(8):465-466
ObjectiveTo observe the effect of early rehabilitation on patients with acute severe brain injury.Methods180 cases with acute severe brain injury were tr eated with early rehabilitation measurements consisted of breath rehabilitation,passive activity of joints in range of joint motion, body posture change,preve ntion of the lower urinary tract infection, and rehabilitation with Traditional Chinese Medicine,etc.Results18 patients (8.3%) co mplicated infection of lungs during the acute stage, no patient died of complica tions.Conclusions The early rehabilitation can pre vent patients with severe brain injury from complications and improve the effica cy of the treatment.
7.Expressions of p75NTR, Bax and Bcl-2 and cell apoptosis in rat cortical neurons following mechanical injury
Haichun LIU ; Tao XIN ; Kaiyun YANG ; Wenliang WU ; Yunzhen CHEN
Chinese Journal of Trauma 2012;28(2):179-183
ObjectiveTo study the expressions of p75NTR,Bax and Bcl-2 and cell apoptosis in rat cortical neurons following mechanical injury and discuss the mechanism and mutual action way during the apoptosis of rat neurons after mechanical injury.Methods Cortical neuron cultures were prepared from the brain tissues of day 17 rat embryos and were exposed to mechanical injury seven days after seeding.After the traumatic neuron injury models were created,the apoptosis ratio of neurons was tested at several time points.The models were divided into the minor,moderate,severe injury groups according to the injury severity and the control group.The expressions of Bax and Bcl-2 in each group were detected by immunohistochemistry method and that of p75NTR by Western-blot.Combining with cellular apoptosis ratio in each group shown after FCM analysis,the correlation between the expressions of p75NTR,Bax and Bcl-2 and the apoptosis in rat neurons after mechanical injury could be analyzed.ResultsThe apoptosis ratio of the neurons in all the injury groups was obviously higher than that in the control group,with significantly higher apoptosis ratio of the neurons in the severe injury group than the minor and moderate injury groups (P <0.05).P75NTR,Bax and Bcl-2 were all expressed in all the injury groups,with statistical differences between groups.The expression of Bax in the severe injury group was significantly higher than that in the minor and moderate injury groups(P < 0.05 ). Conclusions p75NTR expression and Bax/Bcl-2 ratio are closely correlated with neuron apoptosis.The early expression of p75NTR may be one of mechnisms for neuronal apoptosis after neuron injury,when Bax and Bcl-2 may be involved.
8.Effect of penehyclidine hydrochloride pretreatment on NF-κB mRNA expression and SOD activity in lung tissues in rats with acute lung injury induced by LPS
Yanqiu AI ; Wenliang LU ; Baofeng YANG ; Wei ZHANG
Chinese Journal of Anesthesiology 2010;30(6):725-727
Objective To investigate the effects of penehyclidine hydrochloride (PHCD) pretreatment on NF-κB mRNA expression and SOD activity in lung tissues in rats with acute lung injury (ALI) induced by LPS.Methods Thirty-two male SD rats, 2 months old, weighing 230-280 g, were randomly divided into 4 groups (n=8 each): control group (group C), ALI group, low dose PHCD group (group LP) and high dose PHCD group (group HP). ALI was induced by intravenous LPS 5 mg/kg via tail vein. Group LP and HP received intraperitoneal PHCD 0. 3 and 1 mg/kg respectively 30 min before LPS administration. The rats were killed at 6 h after LPS administration. The lungs were removed immediately for determination of W/D lung weight ratio, lung water content, NF-κB mRNA expression, TNF-α and MDA content, and SOD activity and microscopic examination. Results NF-κB mRNA expression, TNF-α and MDA content, W/D lung weight ratio and lung water content were significantly higher, while SOD activity was significantly lower in group ALI, LP and HP than in group C (P < 0.05). NF-κB mRNA expression, TNF-α and MDA content, W/D lung weight ratio and lung water content were significantly lower, while SOD activity was significantly higher in group LP and HP than in group ALI and HP than in group LP (P < 0.05). The LPS-induced changes were mitigated by pretreatment with low and high doses of PHCD in group LP and HP.Conclusion Pretreatment with PHCD attenuates LPS-induced ALI by downregulating NF-κB mRNA expression, decreasing local inflammatory response and enhancing anti-oxidant activity.
9.Comparison of Postoperative Efficacy between Cylindrical Abdominoperineal Resection by Laparoscopy and Traditional Abdominoperineal Resection
Hongbin ZHANG ; Zhengqi WEN ; Zhiqiang WANG ; Jun YANG ; Wenliang LI
Journal of Kunming Medical University 2016;37(5):60-63
Objective To evaluate the efficacy between cylindrical abdominoperineal resection(CAPR)by laparoscopy and traditional abdominoperineal resection(APR). Methods A retrospectively analysis was done in the data of patients with low rectal cancer who underwent APR in our department from January 2010 to September 2015. The patients were divided into two groups,including cylindrical abdominoperineal resection by laparoscopy and traditional abdominoperineal resection. The operation time,intraoperative bleeding,the time of intestinal exhaust as well as the postoperative complications of the two groups were compared. Results There were no statistical differences in the intraoperative bleeding,operation time,the time of intestinal exhaust,postoperative intestinal obstruction and pulmonary infection between the two groups(P > 0.05). However,the differences in the rates of perineal incision infection and tumor intestinal perforation between the two groups were statistically significant (P < 0.05). Conclusion CAPR by laparoscopy is safer,more reliable than traditional APR,which can effectively reduce the rates of postoperative incision infection and tumor intestinal perforation of patients.
10.Advance in research on regulation of sRNAs in bacterial biofilm formation
Xiaofang GAO ; Zizhong LIU ; Wenliang LI ; Ruifu YANG ; Yanping HAN
Military Medical Sciences 2017;41(6):530-533,542
Small RNAs(sRNAs) play a significant role in the regulation of bacterial growth.When sensing certain environmental cues such as fluctuation of nutrient concentration, temperature, pH, and osmolarity, sRNAs can influence the expression of target genes.The formation of biofilms is initiated by bacteria transitioning from the planktonic to the surface-associated mode of growth, which is a self-produced extracellular matrix composed of proteins, polysaccharides, and DNA.Recent evidences have shown that small RNA plays an important role in the regulation of bacterial biofilm formation.sRNAs have key roles in biofilm formation process by base pairing with target mRNAs or interaction with modulating proteins.This review discussed the regulation mechanism and pathway of sRNAs in bacterial biofilms formation, and summarized three classical regulatory models of sRNAs in bacterial biofilms formation, this review also gives the research status and development direction of sRNAs in bacterial biofilms formation.