1.Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect.
Zhen WANG ; Da-chuan LIANG ; Jie-yu BAI ; Ning KANG ; Jun-yu FENG ; Zi-quan YANG
China Journal of Orthopaedics and Traumatology 2015;28(5):433-440
OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.
METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.
RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.
CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering
2.Mechanical trauma induces increase of sensitivity of myocardium to ischemia/reperfusion injury in rats
Yongjin WEN ; Zi YAN ; Jue TIAN ; Jie WANG ; Chengzhang CAO ; Feng LIANG
Chinese Journal of Trauma 2011;27(11):1033-1037
Objective To observe the sensitivity of myocardium to ischemia/reperfusion (I/R) injury in mechanical trauma rats.Methods The mechanical trauma was established by Noble-Collip drum in the rats that were completely randomized into five groups:sham trauma group,trauma group,sham trauma and sham surgery group,sham trauma and I/R group,trauma and I/R group.The rats were subjected to 30 minutes of ischemia and one hour of reperfusion one week after trauma.The left ventricular systolic pressure (LVSP) and the left ventricular maximum rate of pressure rise and fall ( ± dp/dtmax ) were recorded with BL-410 biological signal recording and analysis system.The levels of serum creatine kinase isoenzyme MB (CK-MB) and cardiac troponinⅠ (cTnI) were detected by double antibody sandwich ABC-ELISA technique.At the end of reperfusion,the heart was excised and stained with Evan' s blue dye and triphenyltetrazolium chloride (TTC) to measure the infarct region with Image-Pro Plus 6.0 image analysis software.Results Compared with the sham traumatic I/R group,the cardiac function in vivo was significantly decreased in the traumatic I/R group ( P < 0.01 ).While the serum CK-MB [ (4 960 ± 588 ) ng/ml:(2 925 ± 426) ng/ml,P < 0.01 ],cTnI [ ( 18.10 ± 3.06 ) ng/ml:( 6.67 ±1.57 ) ng/ml,P < 0.01 ] levels and myocardial infarct size [ ( 36.70 ± 7.42 ) %:( 22.27 ± 4.54 ) %,P<0.01] were obviously higher in the traumatic I/R group compared with the sham traumatic I/R group.Conclusion Mechanical trauma increases the sensitivity of myocardium to I/R injury in rats.
3.SAS Application in Studies on Medium Optimizationof Lactic Acid Fennenlation
Zi-Jun XU ; Jian LI ; Feng-Lai LIANG ; Jian-Fang MA ; Ru-Lin LIU ;
Microbiology 1992;0(03):-
The optimum medium for Lactobacillus M7 was systematically studied with SAS system. Firstly, the prime factors affecting lactic acid yield were selected by means of Plackett-Burman design; secondly, the pnme facias were optimized by response rurface analysis. Under the optimum level determined, the yield is increased by 15%.
4.Sky bone expander kyphoplasty for osteoporotic vertebral body compression fractures
Wei-Guo LIANG ; Zi-Qiang ZHOU ; Jing-Feng WU ; Shao-Hui YE ; Wei-Xiong YE ;
Chinese Journal of Trauma 2003;0(08):-
Objective To investigate surgical technique and clinical efficacy of Sky bone ex- pander kyphoplasty in the treatment of osteoporotic vertebral body compression fractures.Methods Eighteen cases with osteoporotic vertebral body compression fractures were treated with Sky bone expander kyphoplasty from August 2004 to November 2005.Under the local anesthesia,3.5-5ml of bone cements were injected into each pathologic vertebral body through unipedicle approach after reduction procedure was done with Sky bone expander.Results The postoperative follow-up ranged from 3 to 11 months, with an average of 4.5 months.Back pain was effectively relieved after the operation in all cases.No complications occurred.Conclusion The Sky bone expander kyphoplasty has the advantages of safe- ty,easy operation,minimal invasion,effective restoration of the vertebral body height and fast relief of pain.
5.Molecular Cloning and Sequencing of HCV E2 Gene
Dong-wei, ZHANG ; Bu-feng, LIANG ; Zi-bai, QI ; Shi-Gan, LING
Virologica Sinica 2001;16(1):40-44
HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.
6.Observation on the efficacy of Conbercept for chronic central serous chorioretinopathy
Liang, YAO ; Sha-Sha, LÜ ; Zi-Yao, LIU ; Hai-Xiao, FENG ; Yu-Ping, ZHENG ; Jian-Ming, WANG ; Feng, WANG
International Eye Science 2017;17(6):1139-1142
AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P<0.01) between those of before and 2mo and 6mo after.The mean BCVA of the prior treatment was 0.53±0.32, the after treatment was 0.65±0.20, there was no different between the two(P>0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.
7.Spectrum-effect relationship of reducing phlegm effect of Peucedanum harrysmithii var. subglabrum.
Jian-di LIANG ; Liang-gong ZHAO ; Xiao-hua LIU ; Wen LI ; Zi-long DANG ; Jin LIANG ; Shi-lan FENG
China Journal of Chinese Materia Medica 2012;37(19):2894-2897
OBJECTIVETo analyze the relationship between high-performance liquid chromatography (HPLC) fingerprints of the chloroform extract fractions of Peucedanum harrysmithii var. subglabrum (PHS) and its phlegm-reducing effect, in order to establish "active component group for reducing phlegm".
METHODHPLC was adopted to determine and analyze HPLC fingerprints of chloroform extract fractions of PHS. Phenol red expectorant experiment was used to observe the phlegm-reducing effect in mice. Mice were administered intragastrically with chloroform extract fractions for 6 days (1.4 g x kg(-1)), with acute bronchitis syrup as the positive control drug (12 mL x kg(-1)). The phenol red secretion in mice was determined by spectrophotometer. Then the grey relational analysis was used to study the spectrum-effect relationship.
RESULTThe phlegm-reducing effect of the chloroform extract fractions of PHS were resulted from the combined effect of all of its chemical components. Its various characteristic peaks represented different chemical components, and the order of their contributions to the phlegm-reducing effect was (number of peaks) 13 > 12 > 16 > 18 > 19 > 6 > 20 > 14 > 1 > 11 > 15 > 10 > 17 > 2 > 5 > 4 > 7 > 3 > 8 > 9, in No. 1, 3, 4, 10, 13 and 16 characteristic peaks were identified as marmesin, psoralen, xanthotoxin, Pd-Ib, pteryxin and peuformosin.
CONCLUSIONThe chloroform extract fractions of PHS show strongly phlegm-reducing effect. There may be certain relationship between their HPLC fingerprint and phlegm-reducing effect.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Ferns ; chemistry ; Mucus ; drug effects
8.The effect of inhibiting EOLA1 expression in ECV304 cells.
Zi-wen LIANG ; Zong-cheng YANG ; Jian CHEN ; Xiao-feng LUO ; Xing-ming WANG
Chinese Journal of Medical Genetics 2007;24(3):293-297
OBJECTIVETo study the effect of inhibiting the expression of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) on proliferation of human umbilical vein endothelial cell line ECV304.
METHODSAfter constructing and transfecting EGFP-EOLA1 fusion protein expressive vector into ECV304 cells, the transfected cells was cultured in M199 containing G418 for 5 weeks to screen the cell line stable expression EGFP-EOLA1 fusion protein. Oligonucleotides targeting EOLA1 at different sites were synthesized and inserted into pSinencer3.1/H1 vector. Then, the recombinant vector was transfected into the cultured ECV304 cells and the inhibiting effect to target gene EOLA1 was investigated by observing the green fluorescence in transfected cells under inverted fluorescent microscope and by Western blot assay. The proliferation of ECV304 cells was numbered when the expression of EOLA1 in ECV304 cells was inhibited by RNA interference.
RESULTSThe ECV304 cell line stably expressing EGFP-EOLA1 fusion protein was constructed and the siEOLA1 interfere vectors can knock down EOLA1 gene expression specially. When blocking the expression of EOLA1 in ECV304 cells,the proliferation of cells slowed down.
CONCLUSIONEOLA1 maybe has a role on the proliferation of cells.
Blotting, Western ; Cell Line ; Cell Proliferation ; Down-Regulation ; genetics ; Humans ; Membrane Proteins ; genetics ; Phenotype ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Recombinant Proteins ; genetics
9.Differential display of homeobox gene expressions in the normal, wounded human fetal and adult skins by DNA microarray.
Hui-Feng SONG ; Jia-Ke CHAI ; Min-Liang CHEN ; Zi-Hao LIN
Chinese Journal of Plastic Surgery 2005;21(5):372-374
OBJECTIVETo explore the differential expression of homeobox genes in the normal, wounded human fetal and adult skins and its significance in fetal scarless healing.
METHODSGene chips containing 14 000 human genes were used to investigate homeobox gene expressions of the normal, wounded human fetal and adult skins.
RESULTSThere were significant differences between the expression of homeobox genes, especially for PRX-2, HOXB13, HOXB6 and HOXB7.
CONCLUSIONSThe homeobox gene is in close relation to developmental biology. The different expressions and changes of homeobox genes in the normal, wounded human fetal and adult skin may be a primary cause of different wound healing between fetal and adult skin.
Adolescent ; Adult ; Animals ; Cells, Cultured ; Child ; Female ; Fetus ; Gene Expression ; Gene Expression Profiling ; Genes, Homeobox ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Oligonucleotide Array Sequence Analysis ; Skin ; Wound Healing ; genetics ; Young Adult
10.Scene-trait coping style of military rescuers in Wenchuan earthquake.
Li-Yi ZHANG ; Xin-Yang SUN ; Ling-Ming KONG ; De-Hua TU ; Liang ZHANG ; Guang-Yao LI ; Zi-Xiang SONG ; Chun-Xia CHEN ; Gao-Feng YAO ; Jie SHI
Chinese Journal of Applied Physiology 2014;30(6):538-543
OBJECTIVEThis study aimed to explore the scene-trait coping style of military rescuers in Wenchuan earthquake in an effort to provide scientific evidences for mental intervention program for Chinese military personnel.
METHODSBy cluster sampling, a total of 151 military rescuers and 331 control servicemen were administered the military personnel scene-trait coping style scale (MPSTCSS).
RESULTSAll active coping factor scores, and passive coping factor scores of affection, health and economy in the rescuer group were significantly higher than those in control group (P < 0.05). The 21-above age subgroup, the 3-year plus service subgroup, and the officer subgroup had significantly higher active coping factor scores on military tasks, military experience and personal development than those of the 21-below age subgroup, 3-year minus service subgroup and the soldier subgroup, respectively (P < 0.05 or P < 0.01).
CONCLUSIONThe earthquake relief servicemen can cope with stressful situations better than control group by taking active coping style. The officers, servicemen older than 21 years, and servicemen with more service duration than 3 years could usually take active coping style.
Adaptation, Psychological ; Disasters ; Earthquakes ; Humans ; Military Personnel ; psychology ; Stress, Psychological