1.Co-expression of human bone morphogenetic protein-2 and vascular endothelial growth factor-165 in human bone marrow mesenchymal stem cells
Guoping WU ; Li TENG ; Lai GUI ; Li GUO ; Kai YANG
Chinese Journal of Tissue Engineering Research 2007;11(3):587-591
BACKGROUND: Bone morphogenetic proteins (BMPs) are involved in the formation of various tissues including bone, cartilage, tendon, and ligament. Vascular endothelial growth factor (VEGF) promotes angiogenesis by increasing the permeability and migration of endothelial cells.OBJECTIVE: To construct a co-expressing vector of human bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165), and observe the expression of BMP2 and VEGF165 in human bone marrow mesenchymal stem cells (hBMSCs).DESIGN: Observation control trail.SETTING: Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College and Plastic Surgery Hospital of Peking Union Medical College, Chinese Academy of Medical Sciences.MATERIALS: The MSCs derived from the healthy adult volunteers of marrow donors. pIRES-EGFP-hVEGF165 containing total length of cDNA sequence of human VEGF165 gene was provided by Dr. Cheng Ting from Plastic Surgery Hospital of Peking Union Medical College. pSP65-hBMP2 containing total length of cDNA sequence of human BMP2 gene was provided by Dr. Guo Ximin from the Academy of Military Medical Sciences. Enkaryotic expression vector pIRESneo (Clontech Company) Pyrobest DNA Polymerae, restriction enzyme, DNA ligase and plasmid extraction kit, DNA Fragment Purification Kit (TaKaRa Company), LiorfectamineTM liposome transfection kit, DMEM medium, trypase, TRIzoIRNA extraction kit (Gibco BRL), Omniscript RT kit (Qiagen), TaqplusDNA polymerase (Promega), PMSF, leupeptin, aprotinin, chymostatin (Sigma), protease inhibitor, PVDF membrane (Amersham-Pharmacia Biotech), rabbit anti-human BMP2 antibody and VEGF monoclonal antibody (Santa Cruz Company), goat anti-rabbit lgG-peroxydase (Wuhan Boster), G418 (Ameresco Company in U.S).METHODS: The experiment was conducted in the Affiliated Hospital of Luzhou Medical College and the Plastic Surgery Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences between June 2005 and April 2006.hBMP2 and hVEGF165 cDNA were directional cloned into multiple clone sites of the eukaryotic expression vector pIRESneo. The recombinant plasmid was identified by restrictive enzyme Xho Ⅰ/Bgl Ⅱ digestion analysis and DNA sequencing. Liposome-mediated gene transfer method was used to transfect hBMSCs. For observation, the transfected cells were divided into IRES-hBMP2-VEGF165 group, pIRES-hBMP2 group, pIRES-VEGF165 group and empty vector group, which were transfected with pIRES-hBMP2-VEGF165, pIRES-hBMP2, pIRES-VEGF165 and pIRES-neo. Meanwhile, the same number nontransfected cells were selected as blank control group. The reverse transcription polymerase chain reaction (RT-PCR) and Weszern blot were employed to observe the expression and secretion of hBMP2 and hVEGF165 gene and protein.expression vector hBMP2 and hVEGF165 gene sequence were the same as reported after restrictive enzyme EcoRI and Bgl Ⅱ digestion analysis and pIRES-BMP2 gene sequencing, which showed that the recombinant plasmid pIRES-hBMP2-VEGF165 highly expressed hBMP2-mRNA and VEGF165-mRNA, but the non-transfected or transfected with pIRES-hBMP2-VEGF165 or pIRES-hBMP2 secreted a great quantity of hBMP2, but that non-transfected or transfected with pIRES-VEGF165 or empty vector secreted only little.CONCLUSION: The co-expressing vector of hBMP2 and hVEGF165 can be expressed stably in hBMSCs.
2.Effect of socioenvironmental therapy on improving emotional distress for heroin addicts accepting compulsory isolation and labour camp rehabilitation
Shishi YU ; Li GUO ; Qinglang ZENG ; Chunqiang LI ; Kai HUANG
Chinese Journal of Behavioral Medicine and Brain Science 2012;(12):1122-1124
Objective To evaluate the psychological intervention ettectiveness ot socioenvironmental therapyfor heroin addicts accepting compulsory isolation.Methods Totally 100 male heroin addicts accepting compulsory isolation and Labour Camp Rehabilitation were selected and randomly assigned to socioenvironmental therapeutic group (ST group,50 cases) and control group (CT group,50 cases).A two-month socioenvironmental therapy was selectively proceeding in ST group by a same psychological team,while CT group still received conventional rehabilitation.During the 6-month follow-up,self-rating anxiety scale(SAS),self-rating depression scal (SDS) and self-made questionnaires were used every 3 months,to statistically estimate psychological status of subjects.Results Before treatment,no statistically significant difference between the two groups;after treatment,SDS score of CT group was 56.16 ± 9.12,and was lower than that in ST group(48.28-± 9.55) (P < 0.01) ; SAS score of ST group was 46.28 ± 7.78,and was lower than that in CT group (50.72 ± 8.32) (P < 0.01).In the 3-month follow-up evaluation,depression,anxiety scores in CT and ST group were significantly lower,the difference was statistically significant (P < 0.01) ; 6 months after treatment,depression,anxiety score showed was significant difference between two groups(P< 0.01).Conclusion Group psychotherapy especially socioenvironmental therapy is quick and long-acting in improving the psychological status of heroin addicts in aspects of depression and anxiety levels,which is distinctly evidenced by the short-term and long-term curative results.
3.Two new sesquiterpenoids from basidiomycete Tyromyces chioneus.
Hua GUO ; Tao FENG ; Zheng-Hui LI ; Ji-Kai LIU
Acta Pharmaceutica Sinica 2014;49(11):1578-1581
Two new sesquiterpenoids, named as tyromols A and B (1 and 2), were isolated from cultures of basidiomycete Tyromyces chioneus, along with two previously reported 15-hydroxy-6 α, 12-epoxy-7β, 10αH, 11βH-spiroax-4-ene (3) and agripilol C (4). Compounds 1-4 were separated and purified by silica gel, RP-18, Sephadex LH-20 column chromatography. Their structures were elucidated on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments.
Basidiomycota
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chemistry
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Magnetic Resonance Spectroscopy
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Sesquiterpenes
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chemistry
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isolation & purification
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Sesterterpenes
4.The effect of cadmium on action potential of ventricular myocytes in guinea pig.
Shun-Kai GONG ; Guo-Bing YU ; Kun LI
Chinese Journal of Applied Physiology 2007;23(3):291-384
Action Potentials
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drug effects
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Animals
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Cadmium
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toxicity
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Guinea Pigs
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Male
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Myocytes, Cardiac
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drug effects
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physiology
5.A correlation study between high resolution CT appearances and expression of transforming growth factor-β,tumor necrosis factor-α in radiation-induced lung injury of rats
Lili GUO ; Guangjun CHENG ; Shaodong LI ; Kai XU
Chinese Journal of Radiology 2008;42(9):984-988
Objective To study the correlation between high resolution computed tomography manifestations and expression of transforming growth factor beta,tumor necrosis factor alpha in radiationinduced lung injury of rats,and to investigate the vslues of cytokine detection and HRCT scanning for the prediction and early diagnosis of radiation-induced lung injury.Methods Forty-eight Sprague-Dawley (SD)rats were randomly divided into eight groups,group A was normal control group.and group BH were irradiated with a single dose of 15 Gy to the lungs.HRCT scanning Wag performed before and 1 week,2,4,8,12,16,24 weeks after radiation in group A-H respectively.The expression of TGF-beta and TNF-alpha were detected with ELISA. All the rats were killed to observe pathological changes of their lungs.HRCT signs,levels of cytokine were simultaneously compared and analyzed.The t-test and Spenrman rank correlation were used for the statistics.Results Four HRCT signs were observed during the 24 weeks after radiation,including ground-glass opacity(1 case),patchy consolidation(8 cases),massive consolidation(7 cases)and fibrosis(3 cases).The average levels of TGF-beta in group B-H[(3.33±0.47),(3.20±0.65),(3.12±0.45),(3.54±0.80),(3.30±1.13),(2.49±0.67),(4.19±0.22)μg/L,respectively]were hisher than the control group[(0.45±0.14)μg/L.P<0.05].At 24 weeks the average level increased to the highest peak[(4.19 4-0.22)μg/L,P<0.05].At 1 week and 2,4,8,12 weeks after radiation,the average level of TNF-alpha in radiation group[(236.52±29.01),(214.91±34.53),(270.97±42.12),(208.83±86.51),(208.83±82.23)ng/L]was hisher than the control group[(31.78±0.92)ng/L,P<0.05].The average level increased to the highest peak [(270.97±42.12)ng/L,P<0.05]at 4 weeks.The average level at 16 and 24 weeks[(60.63±38.49),(32.07±1.05)ng/L]dropped to the level similar to the control group(P>0.05).There were no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha(rs=0.5570 and 0.1013.P>0.05).HRCT signs were correlated with pathological changes.Conciusions The monitoring of TGF-beta and TNF.alpha in the serum after irradiation can predict the development of radiation-induced lung injury.There are no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha.
6.Emergency nursing of the severe complication during the implantation of permanent pacemaker
Fei LI ; Shuang MA ; Lei CHEN ; Kai GUO ; Yaling HAN
Chinese Journal of Practical Nursing 2017;33(1):21-23
Objective To evaluate emergency nursing methods to permanent pacemaker patients with severe complication and acute hemodynamic changes during implantation. Methods The emergency nursing methods were analyzed retrospectively in 9 of 2 027 patients over 6 years, who had serious complication occurred during the pacemaker implantation Results There were 4 patients with pneumothorax, 2 with pericardial tamponed, 1 with ventricular fibrillation and 2 with acute left ventricular failure in 9 patients (0.4%). Observed the changes of symptoms and signs carefully,found the severe complications at the first time and performed the specific nursing based on different kinds of conditions and improved the prognosis of these patients. Conclusions The serious complications during the implantation of permanent pacemaker were life-threatening, and the specific nursing scheme to different types of complications could increase the successful rate of emergency service.
7.Eosinophilic cystitis mimicking muscle-invasive bladder cancer: a case report and literature review
Kai CAO ; Rong YANG ; Xiaogong LI ; Hongqian GUO
Chinese Journal of Urology 2016;37(6):440-442
Objective To evaluate the clinical and pathological features of eosinophilic cystitis (EC),and to discuss its diagnosis and treatment.Methods The clinical and pathological data of one case of eosinophilic cystitis were reported.An 86-year-old male with gross hematuria and urinary urgency for 15 days was admitted on 16 March 2015.A computed tomography scan showed uneven thickening of the bladder wall and a nodular soft tissue on the anterior wall protruding into the bladder cavity.Results Cystoscopic examination showed diffuse thickening of the bladder with polypoid red lesions on the anterior wall.Transurethral biopsies revealed infiltrating eosinophils,which was consistent with the diagnosis of EC.The patient was then treated with anti-inflammatory drugs,corticosteroids and anti-histamine drugs.At 6 months of follow-up,the patient was asymptomatic and had no evidence of local recurrence by ultrasonography.Conclusions Eosinophilic cystitis is a rare disease without specific clinical and imaging features,which can mimic bladder tumor.The final diagnosis of this disease depends on the pathology.Transurethral resection of the lesion along with postoperative hormone and antihistamines administration are the main choices of treatment.
9.Sequence Analysis of 4.0 kb Region Upstream of the Chitinase Gene of Spodoptera litura Nucleopolyhedrovirus
Guo-dong, HU ; Yi, PANG ; Kai, YANG ; Chong-Bi, LI
Virologica Sinica 2001;16(2):161-165
The region of about 4.0 kb upstream of Spodoptera litura nucleopolyhedrovirus (SpltNPV) chiA gene was sequenced, in which six open reading frames(ORF1~6) were found. These ORFs are 156, 297, 540, 369, 1281, and 228 nucleotides long, encoding the proteins of 51, 98, 179, 122, 426, and 75 amino acids with the molecular weight of 6.15 kD, 11.46 kD, 21.70 kD, 14.69 kD, 47.59 kD, and 9.09 kD respectively. One early promoter motif CAGT in ORF1 and ORF3, two early promoter motifs CAGT in ORF2, one late promoter motif TAAG in ORF4 and two late promoter motifs TAAG in ORF5 were found in 5′noncoding regions of these ORFs. The polyadenylation signals, AATAAA, are located downstream of the translation stop codon of ORF1, ORF4 and ORF5. ORF4 is the homologous gene of AcMNPV ORF53, BmNPV ORF42, OpMNPV ORF56 and LdMNPV ORF54. Compared with all genes from baculoviruses, ORF1, ORF2 and ORF6 have no homologous genes. It is suggested that ORF1, ORF2 and ORF6 may be three novel baculovirus genes.
10.Artificial hydrothorax combined with one-lung ventilation to aid percutaneous ablation treatment of liver carcinoma in the hepatic dome
Guanghui GUO ; Erjiao XU ; Rongqin ZHENG ; Qingjin ZENG ; Kai LI
Chinese Journal of Ultrasonography 2015;24(11):959-962
Objective To evaluate the clinical value of artificial hydrothorax combined with one-lung ventilation to aid ablation treatment of liver carcinoma in the hepatic dome.Methods Twenty-one patients with liver carcinoma located in the hepatic dome and affected by the lung gas were enrolled.Double lumen endobronchial intubation anesthesia was used and thoracical tube was used to apply artificial pleural effusion.The lesions'ultrasound image were recorded for the patients without artificial hydrothorax,or with artificial hydrothorax only and artificial hydrothorax combined with one-lung ventilation.Ultrasound images were reviewed and scored as 1-5 according to the clarity and completeness of the lesion.Ablation efficacy and adverse reactions were recorded in the follow-up.Results Twenty-one patients were successfully applied artificial hydrothorax and one lung ventilation.The average normal saline used for artificial hydrothorax were (738± 260)ml.The ultrasound score for not using artificial hydrothorax,using artificial hydrothorax only and artificial hydrothorax combined with one lung ventilation were 1.13 ± 0.35 (1-2),3.00 ± 0.85 (2-5) and 4.53 ± 0.64(3-5),respectively,statistical difference was found between each method(P <0.05).Artificial hydrothorax combined with one-lung ventilation had much higher ultrasound score than the other 2 methods.No complication related to artificial hydrothorax or one-lung ventilation was found.CT/MRI at one month after ablation showed that all the lesions were completely ablated.In the follow-up for 2-14 months on average,1 case of local tumor progression and 1 case of intrahapetic relapse were found.Conclusions Artificial hydrothorax combine one-lung ventilation could effectively enhance ultrasound image for the lesions in the dome whose acoustic window was affected by lung gas.