1.Damaged PC12 cells in Transwell culture system for promoting the transdifferentiation of bone marrow stromal stem cells into neuron-like cells
Jin ZHOU ; Xiaoguang LUO ; Chaodong ZHANG
Chinese Journal of Tissue Engineering Research 2007;11(37):7481-7484
BACKGROUND: Drug treatment has unsatisfactory effect on Alzheimer disease, while many studies have indicated that the transplantation of bone marrow stromal cells (MSCs) is effective on Parkinson disease, cerebral ischemia, etc., but the mechanism is still unclear.OBJECTIVE: To imitate transplantation environment by co-culture of amyloid β1-40 (Aβ1-40) damaged PC12 cells and MSCs, observe the effect of bi-directional information feedback in the microenvironment on the transdifferentiation of MSCs to nerve cells, and observe its protective effect on the apoptosis of damaged PC12 cells.DESTGN: A comparative observation.SETTING: Department of Neurology, China Medical University.MATERIALS: SD rats of 2-3 weeks after birth either male or female were used. PC12 cell lines were purchased from the Institute of Cell Biology, Chinese Academy of Sciences; neuro-specific enolase (1:50, Boster, Wuhan);Methyl-thiazol-tetrazolium (MTT) 15 μL (terminal concentration of 0.5 g/L).METHODS: The experiment was carried out in the Experimental Center (provincial experimental animal center) of China Medical University from June to July in 2004. Bilateral femurs were aseptically removed from 1 SD rat, and MSCs were identified using CD44 antibody immunofiuorescently. PC12 was used to replace nerve cells. The PC12 cells were stimulated by Aβ1-40 then transferred by Transwell. There were 5 groups: Group A: normally cultured PC12 co-cultured with MSCs; Group B: Aβ1-40 stimulated PC12 co-cultured with MSCs; Group C: normal PC12 supernatant+MSCs; Group D: damaged PC12 supernatant+MSCs; Group E: MSCs cultured with common medium 1640.MAIN OUTCOME MEASURES:Routine immunohistochemical staining was performed. NSE positive cells were observed under inverted fluorescence microscope, 10 visual fields (200×) were randomly selected to count the positive cells. MTT metabolic rate was used to detect the proliferation of MSCs in each group. The differences of measurement data were compared using the one-way analysis of variance.fluorescent and bright fields, NSE positive cells appeared as red fluorescence, MSCs were bipolar, multipolar and cone shapes, and appeared as neuron-like forms with dendrite-like structure, and there were extensive connections among some neuron-like cells. The NSE positive rates was obviously higher in group B than groups A, C, D and E (P < 0.01 ).in groups A, C, D and E (F=9.713, P< 0.01).
2.Annexin A1 increases the sensitivity of K562 cell to imatinib.
Kangning LI ; Jing JIN ; Xiaoguang CHEN
Acta Pharmaceutica Sinica 2013;48(6):866-73
Annexin A1 (ANXA1) is a kind of endogenous scaffold protein. Previous research showed that ANXA1 could increase markedly with multiple increase of drug resistance in K562/imatinib cell lines in vitro. Here the stable transfection cell strains K562-pEGFP-N1 which was the native control and K562-pEGFP-N1-ANXA1 which can stably express ANXA1 were established using the Lipofectamine 2000 in order to find whether ANXA1 involved in the drug resistance. Cell growth inhibition experiment via MTT and cell proliferation experiment via MTS showed that K562-pEGFP-N1-ANXA1 cell strain was more sensitive to imatinib than the K562-pEGFP-N1 cell strain, and however the ability of proliferation of K562-pEGFP-N1-ANXA1 cell strain did not change compared with the negative control. Western blotting results showed that the expression of proteins in Annexin family did not change; drug resistance proteins, Bcr-Abl/p-Bcr-Abl (Tyr245), Src family kinase for example, did not change; proteins related with cell proliferation and cell cycle, such as ERK1/2MAPK, p-38MAPK, CDK1 and Wee 1, did not change either in the K562-pEGFP-N1-ANXA1 cell strain compared with the negative control. The co-immunoprecipitation result showed that the interaction between ANXA1 and beta-actin in the K562-pEGFP-N1-ANXA1 cell strain increased markedly. The deduction was that ANXA1 may make the K562-pEGFP-N1-ANXA1 cell strain more sensitive to imatinib due to the increased uptake of imatinib via the increase of ANXA1 and the interaction between ANXA1 and beta-actin in the K562-pEGFP-N1-ANXA1 cell strain in vitro.
3.The co-culture system of MSCs and injured PC12 in vitro could inhibit the apoptosis of PC12
Jin ZHOU ; Xiaoguang LUO ; Chaodong ZHANG
Chinese Journal of Neurology 2005;0(07):-
Objective To establish the co-culture system of marrow stromal cells (MSCs)and A?1-40 injured PC12 in vitro and to evaluate the effect and mechanisms of the system inhibiting apoptosis of PC12 induced by A?1-40.Methods MSCs and PC12 were cultured in vitro and identified by CD44 immunofluorescent staining;PC12 were damaged by A?1-40,and transferred by transwell followed by the classification into 5 groups. PI and Annexin-V co-fluorescent staining was performed,then PC12 apoptosis were detected by flow cytometry and EM;Supernatant was analyzed by enzyme-linked immunoadsordent assay (ELISA) to detected TGF-?,NGF,BDNF,and bFGF. Results About 96% MSCs showed CD44 positive cells. Co-culture group had the lowest rate of PC12 apoptosis(46.17%?8.28%,F=61.637,P0.05). Conclusion The co-culture system of MSCs and A?1-40 injured PC12 in vitro could inhibit apoptosis of PC12 induced by A?1-40. Thus grafted MSCs have the possibility to inhibit neuronal apoptosis by A?in the diseased brain.
4.Application of Nutritional Risk Screening 2002 and nutrition support in critically ill patients in a Beijing-based hospital
Sanli JIN ; Xiaoguang LI ; Qian LU
Chinese Journal of Clinical Nutrition 2012;(6):351-354
Objective TO investigate the application of Nutritional Risk Screening 2002 (NRS 2002) and nutrition support in critically ill patients.Methods Totally 172 adult critically ill patients with a hospital stay equal to or above 3 days were consecutively enrolled from a Beijing-hospital from September 2011 to December 2011.Nutrition support was evaluated on a daily basis.NRS 2002,body mass index (BMI),and Acute Physiology and Chronic Health Evalnation Ⅱ (APACHE Ⅱ) were applied to assess the nutritional risks and nutrition status within 24 hours after admission.Results Among all the patients,the rates of nutrition support were 39.5% and 67.9% on the 1st and 7th day respectively.The rate of parenteral nutrition (33.6% ~ 39.5%) was higher than that of enteral nutrition (4.1% ~ 16.0%).NRS 2002,APACHE Ⅱ score,and BMI showed that 160 patients (93.0%) required nutrition support,but only 103 patients (64.4%) actually received nutrition support,among whom 92 (89.3%) were supplied with nutrition support within 3 days after admission.Conclusions The application of nutrition support in critically ill patients is insufficient in this hospital.Nutrition support is typically provided within 3 days after admission.Parenteral nutrition remains the most common type of nutrition support.
5.Feasibility of nanoparticle of Fe2O3 combination with microwave for ablation of renal VX2 tumor
Ziyi GUO ; Xiaoguang LI ; Zhengyu JIN
Chinese Journal of Radiology 2016;50(6):464-468
Objective To evaluate the application of embolic material, iron oxide nanoparticles?lipiodol?gelatin in the improvement of therapeutic effect of microwave ablation in vitrogelatin phantom model and in vivo VX2 renal tumors model. Methods After preparing new embolic material containing nano Fe2O3, in vitro model experiment was firstly conducted to verify the temperature distribution of the microwave ablation. Further verification was performed in bilateral renal VX2 tumors in rabbit model. A total of 10 (20 kidneys) New Zealand White rabbits bearing renal VX2 tumors were treated by renal artery embolization and CT guided microwave ablation, in which left kidneys were treated by material containing nano Fe2O3 in iodized oil and right kidneys were treated by renal embolization with super liquid iodized oil as a control. The same microwave ablation time and power were used for both kidneys. CT perfusion and histopathological examinations were performed to determine the renal perfusion parameters, tumor histopathology and tumor?related angiogenesis. Results In vitro experiments, when nano Fe2O3 was present in the local area at 1cm from the microwave needle at 20 and 60, the temperature reached(59.1 ± 1.7)° C, (95.5 ± 4.5)° C vs.(31.1 ± 2.5)and(45.6 ± 3.5)° C in the controls. Our present results strongly suggested that local nano Fe2O3 particles absorbed microwave and shielded microwave energy transferring to the surrounding tissue. Before and immediately after renal VX2 tumor embolization, blood flow significantly reduced from (31.4 ± 10.6) to (6.1 ± 9.9) ml · 100 ml-1 · min-1. Our pathology findings indicated nano Fe2O3?lipiodol-gelatin embolic agent stored in endovascular regions, while no penetration into the renal parenchyma happened. Conclusions This in vitro model and in vivo experiment suggest that the new embolic material containing nano Fe2O3 may help to improve heat efficiency of MWA microwave, and it is feasible to administer it through transcatheter arterial embolization approach for the microwave ablation therapy of kidney cancer.
6.Efficacy of quadriceps femoris fasciculation induced by low-current nerve stimulation when used to assist ultrasound-guided lumbar plexus block
Ke SUN ; Mei JIN ; Xiaoguang ZHANG
Chinese Journal of Anesthesiology 2017;37(6):700-703
Objective To evaluate the efficacy of quadriceps femoris fasciculation induced by lowcurrent nerve stimulation when used to assist ultrasound-guided lumbar plexus block.Methods One hundred patients of both sexes,aged 18-45 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,weighing 50-85 kg,scheduled for elective unilateral knee arthroscopy,were selected and randomly divided into 2 groups (n =50 each) using a random number table:ultrasound assisted by nerve stimulator group (group SU) and ultrasound group (group U).The shamrock method was used to perform the ultrasound-guided lumbar plexus block in two groups.In group SU,the nerve stimulator with current 0.35 mA and frequency 1 Hz was used in the process of puncture,and 0.5% ropivacaine 0.4 ml/kg was administrated when quadriceps femoris fasciculation was induced.In group U,when the tip of the nerve stimulating needle was located around the lumbar plexus,which was confirmed by ultrasound,0.5% ropivacaine 0.4 ml/kg was administrated.The time of puncture,depth of needle insertion,onset time of block and effective block were recorded.Motor block was assessed using the modified knee score,and the development of complications was recorded within 24 h after block.Results Compared with group U,the onset time of block was significantly shortened,the rate of effective block was increased,the degree of motor block was aggravated (P<0.05),and no significant change was found in the time of puncture or depth of needle insertion in group SU (P>0.05).No complications were observed in two groups.Conclusion Low-current (0.35 mA) nerve stimulation-induced quadriceps femoris fasciculation when used to assist location can improve the efficacy of ultrasound-guided lumbar plexus block.
7.Low Tube Voltage Multi-slice Spiral CT in Evaluation of Malignant Obstruction in Metal Biliary Stents
Ziyi GUO ; Xiaoguang LI ; Zhengyu JIN
Chinese Journal of Medical Imaging 2015;(11):801-803,807
Purpose In order to find out a means that can both meet clinical requirements and reduce radiation dosage in the diagnosis of malignant biliary obstruction, the study aims to evaluate the image quality and the radiation dose of low tube voltage enhanced CT (100 kVp) in displaying malignant biliary obstruction in metal stents. Materials and Methods Between January 2012 and March 2014, 9 patients with highly suspected recurrent occlusion after biliary stent procedure were admitted in Peking Union Medical College Hospital and underwent high resolution three-phase dynamic enhanced CT scan. The CT images on portal veinal phase were reconstructed through the curved planar reformation and the CT values of 184 slices were evaluated along the pathway of the biliary stent. The reconstructed image quality was analyzed when the tube voltage was 140 kVp and 100 kVp respectively, and the different tumor-to-liver contrast-to-noise ratio (CNR) for each lesion, mean image noise, and figure of merit (FOM) between the two data sets were also compared. Results As the tube voltage decreased from 140 to 100 kVp, image noise increased from (19.88±3.62) HU to (24.22±4.24) HU (P<0.001), CNR increased by 15% and FOM improved by 50%. Conclusion A low tube voltage CT technique (100 kVp) improves tumor-to-liver CNR and significantly reduces radiation dose, therefore it is valuable in the manifestation of malignant obstruction in metal stents.
8.An experiment on focal cerebral ischemia reperfusion model in mice established with suture method
Xiaoguang WU ; Yong YAN ; Qian XU ; Hongwei JIN
Chinese Journal of Tissue Engineering Research 2005;9(41):128-130
BACKGROUND: Techniques in the focal cerebral ischemia model in mice have been well developed in some foreign countries. However, rats are commonly used in cerebral ischemia studies due to resource availability in China.OBJECTIVE: To establish reliable and reproducible focal cerebral ischemia reperfusion model in mouse to explore the molecular mechanism of cerebral ischemia reperfusion process in genetic level.DESIGN: Randomized controlled study.SETTING: Fundamental Medical Research Institute of Chengde Medical College. MATERIALS: The experiment was carried out in Fundamental Medical Research Institute of Chengde Medical College from September 2002 to May 2003. Totally 30 Kunming mice (25 to 30 g), provided by Experimental Animal Center, Chengde Medical College, were randomly grouped as follows: control group, 6, 3-hour occlusion and 18-hour, 24-hour reperfusion, with 6 mice in each group respectively.INTERVENTIONS: Nylon thread (qb0.128 mm) was soaked in paraffin wax and inserted into internal carotid artery in mouse to reversibly occlude the middle cerebral at the right side after 3 hours, then was removed, and reperfusion was performed in 18, 24 hours, respectively.MAIN OUTCOME MEASURES: The infarct volume was observed and neurological deficit scored was analysis.of mice in operation group was shown as head turning to left, adduction of left forelimb, internal rotation of shoulder, decrease of muscular tension of left forelimb, unable to straighten completely during tail lifted. At independent activity, mice turned to left as the left hidlegs as the center of a circle; under the abdomen, left forelimb was crossed with right forelimb as scissor shape. Palsy of left limbs was observed obviously, especially forelimb.tion was shown clearly with TTC. The infarct volumes were (13±2.3) mm3 and (16±4.5) mm3 for 18 hours and 24 hours reperfusion and 3-hour ischemia, respectively. Pathological changes were observed under light microscope after ischemia. The infarct volume increased as the increasing of reperfusion time, including 3-hour blocking and 18-hour and 24-hour reperfusion.CONCLUSION: Using suture method to establish focal cerebral ischemia reperfusion model in mice, it is less invasion and trauma without requiring skull opening. Ischemia position is relatively fixed, thus, ischemia reperfusion time can be controlled precisely. It is also an ideal animal model to study pathological changes in cerebral vascular diseases and an efficient tool to study the therapeutic effect of medicine after surgery.
9.CT-guided transgluteal approach for drainage of deep pelvic abscesses
Xiaoguang LI ; Zhengyu JIN ; Haifeng SHI ; Yumei LI
Chinese Journal of Radiology 2008;42(7):749-751
Objective To evaluate the safety and efficacy of CT-guided transgluteal approach for drainage of deep pelvic abscesses. Methods From April 2000 to August 2007, 12 patients with deep pelvic abscesses underwent CT-guided transgluteal drainage. The clinical data, number of drainage cathete rsplaced, amount of drain, duration of placement, complications and follow-up were retrospectively analysed.Results One drainage catheter was successfully placed for each patient. The amount of abscess drained was30 to 180 ml (mean 52 ml) and the duration of placement was 6 to 34 days (mean 11 days). No major complications occurred. Conclusion The transgluteal approach to the drainage of deep pelvic abscess under CT guidance was safe and effective.
10.Transcatheter arterial embolization for the treatment of splenic artery aneurysms: clinical results in eight cases
Zhiwei WANG ; Xiaoguang LI ; Zhengyu JIN ; Ning YANG
Journal of Interventional Radiology 2009;18(11):871-873
Objective To assess the feasibility and effectiveness of transcatheter arterial embolization for the treatment of splenic artery aneurysms (SAAs). Methods Transcatheter splenic arterial embolization with coils was performed in eight cases of SAAs. The patients were followed for 3-36 months. The follow-up examinations with CT or color Doppler ultrasonic angiography were made. Results Technical success was achieved in all eight cases with no severe complications. During the follow-up period, one patient died of severe abdominal bleeding 10 months after the operation and the other seven patients remained in good condition with no occurrence of re-canalization of the lesions. Conclusion Transeatheter arterial embolization is a safe and effective treatment for splenic artery aneurysms. It is necessary to make a regular follow-up imaging study.