1.Effects of stromal cells derived factor-1/CXCR4 on migration of human bone marrow mesenchymal stem cells toward ischemic brain region
Jie ZHU ; Zhujuan ZHOU ; Zili GONG ; Jian ZHENG
Chinese Journal of Tissue Engineering Research 2007;0(19):-
BACKGROUND:Recent research has shown that transplanted bone marrow mesenchymal stem cells(BMSCs) migrate to the injured regions and exert their therapeutic effects in cases of intracranial trauma, stroke, inflammation and degenerative disease.The specific mechanisms involved in their migration to lesions are still to be fully elucidated.OBJECTIVE:To explore the effects of stromal cells derived factor-1(SDF-1) and its receptor CXCR4 on the migration of transplanted BMSCs to ischemic brain lesions.DESIGN, TIME AND SETTING:The cytological in vivo study was performed at the Central Laboratory, Xinqiao Hospital, Third Military Medical University of Chinese PLA from February 2008 to February 2009.MATERIALS:Bone marrow samples were obtained from normal or primary affection non-involved bone marrow patients aged 15-40 years at the Department of Hematology, Xinqiao Hospital, Third Military Medical University of Chinese PLA.A total of 72 healthy male Sprague Dawley rats aged 3-4 months were supplied by the Experimental Animal Center, Research Institute of Surgery, Third Military Medical University of Chinese PLA.METHODS:Human BMSCs were isolated by combination of gradient centrifugation and different adherent time method.The transient middle cerebral artery occlusion(MCAO) was induced using intraluminal vascular occlusion in 54 rats, based on the method described by Nagasawa et al.The remaining 18 rats served as sham operation group, only inserted with thread for 10 mm depth.At 2, 4 and 8 days after cerebral ischemia, the expression of SDF-1 in the ischemic brain was determined by real time RT-PCR and immunohistochemistry in 9 rats from either group.The remaining 36 rat models of cerebral ischemia/reperfusion were equally and randomly assigned into a cell transplantation and solution control groups.1 mL human BMSCs(2?109/L cells) or 1 mL phosphate buffered saline were slowly infused through the caudal vein at 24 hours following reperfusion.MAIN OUTCOME MEASURES:CXCR4 mRNA and protein expression in human BMSCs was determined.SDF-1 mRNA and protein expression following ischemia/reperfusion were detected.Migration of transplanted human BMSCs into the damaged region was observed through immunohistochemistry.RESULTS:RT-PCR showed that human BMSCs were positive for CXCR4 mRNA.Immunocytochemistry revealed that CXCR4 mainly expressed in cell membrane and cytoplasm of human BMSCs.At 2, 4 and 8 days following cerebral ischemia/reperfusion, SDF-1 mRNA levels showed an increased tendency, and showed significant difference compared with the sham operation group(P
2.Effects of hypoxia on chemokine receptor CXCR4 and CX_3CR1 expression in human bone marrow mesenchymal stem cells
Jie ZHU ; Zhujuan ZHOU ; Zili GONG ; Jian ZHENG
Chinese Journal of Tissue Engineering Research 2007;0(23):-
BACKGROUND: Transplanted bone marrow mesenchymal stem cells (BMSCs) migrate to the injured regions and exert their therapeutic effects. The specific mechanisms involved in their directional migration to lesions remain unclear. OBJECTIVE: To investigate the chemokine receptor CXCR4 and CX3CR1 expression of human BMSCs in hypoxia culture. DESIGN, TIME AND SETTING: The cytology in vitro study was performed at the Central Laboratory, Xingqiao Hospital, Third Military Medical University of Chinese PLA from February 2008 to February 2009. MATERIALS: Cells harvested from the iliac heparinized bone marrow were obtained by iliac crest aspiration from healthy adult volunteers, aged 15 to 40 years old, at the Department of Hematology, Xinqiao Hospital, Third Military Medical University of Chinese PLA. METHODS: Bone marrow was obtained by puncture. Human BMSCs were harvested by combination of density and gradient centrifugation and different adherent method. Cells at passage 3 were incubated in a 25 cm2 flask. When 70%-80% confluence was found, cells were incubated at 37 ℃ and saturated humidity in an incubator containing 3% O2, 5% CO2, 92% N2 for 48 hours, and those incubated under normal oxygen as controls. MAIN OUTCOME MEASURES: Morphology was examined by phase contrast microscopy. Cell surface markers were tested by flow cytometer. The CXCR4 and CX3CR1 mRNA expression were detected by real-time PCR. The CXCR4 and CX3CR1 protein expression were determined by Western blot assay. RESULTS: All of the cells had a fibroblast-like morphology cultured in vitro and reached 90% confluence at 12-14 days, with the presence of polarity arrangement and whirlpool-shape. Cells were uniformly positive for CD105 (99.38%) and CD29 (99.13%), but negative for CD14 and CD45. Exposure of BMSCs to 3% O2 increased expression of the CXCR4 mRNA and CX3CR1 mRNA, which were respectively 2.130 times and 2.361 times of normal culture; expression of the CXCR4 protein and CX3CR1 protein was respectively 1.69 times and 1.93 times of normal culture. CXCR4 and CX3CR1 mainly expressed in membrane and cytoplasm of human BMSCs. CONCLUSION: Hypoxia (3% O2) can upregulate the expression of CXCR4 and CX3CR1 in human BMSCs, which might be one of the machenisms underlying the migration of BMSCs.
3.A study on the cause of injury and prognostic factors for visual acuity in patients with traumatic optic neu- ropathy
Jiao ZHENG ; Ping WANG ; Qinguo YANG ; Jin GONG ; Yan JIAN ; Junling WANG ; Linglin WANG
Chinese Journal of Nervous and Mental Diseases 2014;45(2):71-74
Objective To analysis the cause of injury of traumatic optic neuropathy and explore the main factors affecting the prognosis of visual acuity. Methods We retrospectively collected clinical data of 104 cases (108) of traumat-ic optic neuropathy from January 2007 to December 2012 in the Renhe Hospital of Three Gorges University. We then ex-amined the cause of injury and analyzed risk factors for poor prognosis of visual acuity in traumatic optic neuropathy us-ing multivariate Logistic regression analysis. Results The mean age of patients was 33.6 ± 12.8 years and 95 cases (91.3%) were males. The injury was mainly caused by non-motor vehicle and motor vehicle accidents (84 cases, 80.8%). Most of the patients came to the hospital within 3 days after the injury (81 cases, 77.9%). Overall efficacy rate of treat-ment was 45.4%(49/108). The effective rate was 29.9%(20/67) for visual acuity without light perception and 70.7%(29/41) for visual acuity with light perception, respectively. Logistic regression analysis showed that the time to treatment≥24h, orbital hemorrhage or orbital fracture, ethmoid or sphenoid sinus bleeding and no light perception vision were an in-dependent prognostic factors. Conclusion Time to treatment time and severity of injury are the independent risk factors for poor visual prognosis of traumatic optic neuropathy whereas controlling these risk factors has important clinical signifi-cance to the treatment and prognosis of traumatic optic neuropathy.
4.Comparison of biological characteristics of mesenchymal stem cells derived from bone marrow, peripheral blood and cord blood
Youzhang HUANG ; Jianliang SHEN ; Lizhong GONG ; Wenjie YIN ; Yi LIU ; Hai CHENG ; Peihao ZHENG ; Jian CEN
Chinese Journal of Tissue Engineering Research 2009;13(45):8966-8970
BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P < 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.
5.Clinical application of CT-guided ~(125)I seed interstitial implantation for recurrent rectum carcinoma
Zhangmin WANG ; Gang HUANG ; Kemin CHEN ; Jian LU ; Ju GONG ; Yunfeng ZHENG ; Liyun ZHANG
Journal of Interventional Radiology 2009;18(9):681-684
Objective to explore the safety and clinical efficacy of CT-guided radioactive seed implantation in treating recurrent rectum carcinoma. Methods CT-guided ~(125)I radioactive seed implantation was carried out in 20 patients with recurrent rectal carcinoma. Treatment planning system was used preoperatively to reconstruct three dimensional image of the tumor and to calculate the estimated seed number and distribution. The tumor matched peripheral dose (MPD) of the radioactive seeds was 80-130 Gy. The radioactivity of the seeds was 0.5-0.8 mCi/seed and the median implanted seeds was 48 (range 25-95) in number. CT scan was made immediately after the implantation to check the quality of the seeds. Change of pain score, tumor size and complications were recorded during the follow-up period. Results Twenty cases composed of 12 males and 8 females, aged 38 to 78 years (median age of 62 years). The follow-up period lasted 2-28 months. On an average, 3 to 7 days after the procedure patients experienced significant pain relief. CT scan performed 2 months after the procedure revealed that complete relief (CR) of the tumor was seen in 2 cases, partial relief (PR) in 13 cases, no change (NC) in 3 cases and progression (PD) in 2 cases. The total effective rate (CR + PR) was 75%. The median survival time was 18.8 months. The survival rate of 1 and 2 years was 75% and 25% respectively. Two cases died of tumor deterioration and 3 cases died of extensive metastases. No complications such as frequent micturation, pain on urination and hematuria occurred during the follow-up period. Conclusion CT-guided ~(125)I radioactive seed implantation is a safe and effective interventional treatment for recurrent rectal carcinoma with reliable short-term efficacy and excellent anti-pain effect.
6.Clinical efficacy of CT-guided ~(125)I seed implantation therapy for advanced pancreatic cancer
Zhongmin WANG ; Gang HUANG ; Kemin CHEN ; Jian LU ; Ju GONG ; Yunfeng ZHENG ; Liyun ZHANG
Journal of Interventional Radiology 2009;18(9):668-672
Objective To discuss the clinical efficacy of CT-guided radioactive ~(125)I seed implantation treatment for unresectable pancreatic cancer. Methods Forty patients with inoperable pancreatic cancer were enrolled in this study, including 25 males and 15 females with an median age of 69 years (38-89 years). Treatment planning system (TPS) was used to reconstruct 3-dimensional images of pancreatic tumor and to define the quantity and distribution of ~(125)I seeds. The radioactivity of ~(125)I seeds was 0.5-0.8 mCi/seed. The seeds were implanted into pancreatic tumor under CT guidance at intervals of 1 cm and were kept away from vessels, pancreatic duct and other adjacent important organs. The tumor matched peripheral dose (MPD) was 60-140 Gy. The median amount of implanted ~(125)I seeds was 36 (18-68) in number. CT scan was performed immediately after the procedure to check the quality of the seeds. In addition, 10 patients received concurrent chemotherapy with arterial infusion of gemcitabin and 5-fluororacil (5-Fu) for 3 to 4 therapeutic courses. Results The median diameter of the tumors was 4.9 cm. The follow-up period was 2 to 28 months. After the treatment the refractory pain was significantly relieved (P < 0.05), and Karnofsky score was dramatically increased (P < 0.05). Most patients experienced relief of pain within 2-5 days after implantation. Two months after treatment, on CT scans the tumors showed completed relief (CR) in 3 cases, partial relief (PR) in 20 cases, no change (NC) in 14 cases and progression (PD) in 3 cases. The overall effective rate (CR+PR) was 57.5%. The median survival time for all patients was 10.2 months, while it was 14.7 months, 10.9 months and 7.1 months for patients in stage Ⅱ, stage Ⅲ and stage IV respectively. For patients in stage Ⅱ, stage Ⅲ and stage Ⅳ, the 6-month cumulative survival rate was 100%, 88% and 62% respectively, while the 12-month cumulative survival rate was 70% , 41% and 0% respectively. After the therapy, liver metastasis occurred in 5 cases and chemoembolization was employed. In three patients, immigration of four radioactive seeds to the liver was found. No serious complications, such as upper GI bleeding, pancreatitis, pancreatic fistula formation and radiation colitis, occurred during the follow-up period. Conclusion CT-guided radioactive ~(125)I seed implantation is a safe, effective and minimally-invasive brachytherapy for unresectable pancreatic cancer with reliable short-term efficacy. It has an excellent anti-pain effect. The curative results can be further improved when chemotherapy is employed together. However, its long-term efficacy needs to be observed.
7.Research on quality system management of software manufacturers for medical devices.
Chinese Journal of Medical Instrumentation 2005;29(4):292-294
Based on the investigation and analysis of the current situation of medical devices' software manufacturers an exploration of new administrative modalities for the enterprises is presented in the paper.
Equipment and Supplies
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standards
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Industry
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organization & administration
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Quality Control
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Software Design
9.Study of a serum protein fingerprint diagnostic model in patients with hypertensive disorder complicating pregnancy
Li-Yun GONG ; Zhen-Yu ZHANG ; Yan-Hua ZHENG ; Jian-Zhong ZHANG ;
Chinese Journal of Obstetrics and Gynecology 2000;0(12):-
Objective To analyze the alterations of serum protein fingerprint in patients with hypertensive disorder complicating pregnancy(HDCP),screen serum biomarker and establish a diagnostic model of HDCP.Methods Surface-enhanced laser desorption lionization-time of flight-mass spectrometry (SELDI-TOF-MS)technology was used to analyze serum including 25 cases of HDCP patients and 30 cases of age-,gravity-and parity-matched healthy term pregnant women on IMAC3-Cu proteinchip before delivery. Biomarker Wizard and Biomarker Pattern software was used to detect protein peaks significantly different between HDCP and controls,and establish a primary diagnostic model of HDCP.This model was further evaluated by blind test using other 16 parts of serum protein fingerprint.Results Ten protein peaks were significantly different at the molecular range of 2000-50 000(P
10.Early therapy monitoring of 125Ⅰ seed interstitial implant in a pancreatic cancer xenograft by 18FFDG Micro-PET/CT
Zhongmin WANG ; Yu LIU ; Kemin CHEN ; Jian LU ; Ju GONG ; Yunfeng ZHENG ; Liyun ZHANG ; Fenju LIU
Chinese Journal of Radiological Medicine and Protection 2011;31(3):256-259,289
Objective To investigate the application value of early evaluation and monitoring of 125Ⅰ interstitial implantation in a pancreatic cancer xeuograft.Methods Xenograft models were created by subcutaneous injection of Sw 1990 human pancreatic cancer cell suspensions into the right hind limbs of the immunodeficient BABL/c nude mice.The tumors size were about 8-10 mm after two weeks.The mice were randomly divided into 3 groups,including control group (n = 4) ,empty seed implantation group (n = 4)and 125Ⅰ implantation group (n = 4).Before treatment and one week after treatment,18F-FDG Micro-PET/CT scan was performed and then maximum standardized uptake values (SUVmax),mean standardized uptake values (SUVmean),tumor size and necrosis rate were measured.HE staining and TK1 immunohistochemistry examination were carried out in the paraffin-embedded sample.Results Before treatment the SUVmax and SUVmean values of three groups did not reach statistical significance.One week after treatment the SUVmax and SUV values of three groups were 3.53 + 1.20 and 0.57±0.26 vs.3.83±2.13 and0.59 ±0.24vs.0.29±0.23 and0.016±0.001,respectively,with a significant difference (F =7.62,P =0.01 ; F = 10.34,P =0.005).The SUVmax and SUVmean values of 125Ⅰ implant group were significantly lower than empty seed implant group and control group and were significantly lower than before treatment.Before treatment,tumor necrosis rate of three groups were not significantly different.Immunohistochemical staining found the TK1 positive staining index of three groups were respectively (64.25±1.71) % ,(62.25±2.22) % and (38.25±1.71) % with statistically significant difference (F =233.67,P < 0.001).The TK1 positive staining index of 125Ⅰ implant group was significantly lower than empty seed implant group and control group.The SUVmax values had some positive correlation with TK1 positive staining index (r = 0.85,P = 0.001).Conclusions 18F-FDG Micro-PET/CT may be useful as a noninvasive imaging modality to assess early response to 125Ⅰ seed brachytherapy in a pancreatic cancer xenograft.