BACKGROUND: The studies in recent years proved that the inflammatory reaction is of the main reasons in the damage of cerebral ischemia reperfusion. The nuclear factor κB (NF-κB), as a kind of transcription factor, plays an important role in regulating the expressions of various inflammatory cell factors in the inflammatory reaction of cerebral ischemia reperfusion. The previous experiments show that puerarin functions to resist the oxidated free radicals and the apoptosis of nerve cells. In case it has the functions of anti-inflammation, its brain protection can be explained further.OBJECTIVE: To study the effect of puerarin on NF-κB for the rats with the damage of ischemia reperfusion.DESIGN: A random parallel controlled study.SETTING: The Emergency Department of Beijing Hospital, Emergency Department of Tongji Hospital, Pathology Department and Experimental Animal Center of Tongji Medical College, and Health Statistics Department of Public Health College of Huazhong University of Science and Technology.MATERIALS: The experiment was started on April 12, 2003 in the Pathology Department of Tongji Medical College. The 75 healthy and clean Wistar rats were randomized into 3 groups with 25 in each, Sham operation group, cerebral ischemia reperfusion group, and puerarin group. Each group was reperfused at 2, 6, 12, 24, and 48 hours after ischemia and 5rats were used at each time point.METHODS: [1] Sham operation group: Without electric coagulation of bilateral vertebral arteries, without blockage of bilateral common carotid arteries, without medicinal administration. [2] Cerebral ischemia reperfusion group: Ten minutes after the blockage of bilateral common carotid arteries with non-invasive artery clamp, the reperfusion was given. At the beginning of reperfusion, the abdominal injection of normal saline 1 mL was applied and later every 6 hours the injection was repeated once. [3] Puerarin group:The procedure was the same as for the reperfusion group, only with normal saline changed to puerarin 100 mg/kg.MAIN OUTCOME MEASURES: At the time points of 2, 6, 12, 24, and 48 hours after reperfusion, the activity of NF-κB and inhibitory protein κB(IP-κB) in the hippocampus CA1 region was examined with immunohistochemical method; the expression of tumor necrosis factor-α (TNF-α) mRNA was measured with in situ hybridization method; and the number of surviving neurons was detected with hematoxylin-eosin (HE) staining.RESULTS: After supplement, 75 rats entered the result analysis. [1] Activity of NF-κB: In the ischemia reperfusion group, it was obviously increased at 2 hours after reperfusion, to the highest at 6 hours, and still higher than that of the sham operation group, (P ＜ 0.01). In the puerarin group, it was lower at each time point than that of the ischemia reperfusion group (P ＜ 0.01). [2] Expression of TNF-α mRNA: In the ischemia reperfusion group, it was obviously increased at 2 hours after reperfusion, to the highest at 12 hours, and still higher than that of the sham operation group at 48 hours (P ＜ 0.01). In the puerarin group, it was lower than that of the ischemia reperfusion group at 6-48 hours (P ＜ 0.01). [3] Activity of IP-κB:In the ischemia reperfusion group, it was obviously decreased at 2 hours after reperfusion, to the lowest at 6 hours, and then gradually increased to the level of 12 hours. In the puerarin group, it was higher than that of the ischemia reperfusion group at each time point (P ＜ 0.01 or 0.05). [4] Number of surviving neurons: In the ischemia reperfusion group, it was decreased gradually with the time prolonging after reperfusion (P ＜ 0.01). In the puerarin group, at each time point, it was higher than that of the ischemia reperfusion group (P ＜ 0.05 or 0.01).CONCLUSION: In the cerebral ischemia reperfusion, puerarin can protect the brain through decreasing the degradation of IP-κB, the activity of NF-κB, the expression of TNF-α mRNA, and the inflammatory reaction.

Background and purpose:Cervical conization, including high frequency loop electrosurgical excision procedure(LEEP) has been widely used in the treatment of cervical diseases, but how to deal with the patients with pathological positive margin is a problem for clinicians.The purpose of this study was to discuss the option of adjuvant treatment after cervical conization with positive margins for patients with cervical neoplasm.Methods:The data of 528 patients who had cervical conization from 1998 to 2008 was reviewed, among which 54 patients with pathological positive margin was retreated and analyzed.Results:Fifty-four patients were divided into observation group and treatment group, 17 cases were in observation group and 37 cases in treatmeat proup.The recurrence / duration / progress rate was 17.6%(3/17), in treatment group it was 2.7%(1/37) in observation.CINⅠ-Ⅱ positive margins in both group had no recurrence;among 14 patients with CINⅢ, 1 lesion persisted, and 1 progressed to cervical squamous cell carcinoma, none in treatment group was recurrent;For those 10 patients with micro-invasive margin-positive cases, 1 progressed to squamous cell carcinoma, the remaining 9 cases were followed up for 26 months without recurrence after operation.One case in invasive cancer group had recurrence.Conclusion:The patients with CINⅢ margin-positive patients after conization should receive individualized treatment.The patient with microinvasive carcinoma should be retreated with either re-conization or hysterectomy;if with margin-positive CINⅢ after conization, or re-conization or directly treated according to guideline addressing for ⅠB, if margin showed microinvasive carcinoma.The patients with margin-positive invasive carcinoma after conization should be treated according to guideline.

To observe the cerebral activating effects of needling at Waiguan (SJ5) versus SJ5 plus Yanglingquan (GB34) points in young healthy volunteers based on the hypothesis of "needling effect of combined acupuncture points relates to the brain activation".

Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).

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Objective To compare two sources of mesenchymal stem cells ( MSCs) from human placenta and umbilical cord, and to optimize a technical solution for bench or clinical studies of MSCs.Methods MSCs were isolated from human placenta and umbilical cord and expanded for analysis.The cell morphology was observed under invert microscope, the immunophenotypic feature of MSCs was analyzed with flow cytometer, the cell proliferation ability was determined by cell cycle assay and cell doubling time, the cell differentiation potential was evaluated by osteogenic and adipogenic induction in vitro as well.Results Both sources of MSCs were adherent cells and exhibited fusiform and fibrous morphology. Furthermore, both MSCs high expressed CD90 and CD105, and were negative for the markers of CD34, CD45 and HLA-DR.The population doubling time of MSCs form human placenta and umbilical cord was 39.5 h and 40.8 h separately, and the results of cell cycle analysis showed that the percent of the two sources of MSCs in G0/G1 phase was 52.12%and 57.50% respectively. The above results demonstrated that both sources of MSCs possessed the similar biological characteristics in morphology, phenotype and as well as proliferation ability.In addition, both of them could be induced into osteoblasts and adipocytes in vitro.Conclusion MSCs from human placenta have the similar biological characteristics to these from human umbilical cord, and both of them are better candidates for bench and clinical research.

BACKGROUND:Human umbilical cord mesenchymal stem cells with capabilities for self-renewal and multi-differentiation have attracted widespread attention. OBJECTIVE:To develop an efficient method for isolation and culture of human umbilical cord mesenchymal stem cells, and to analyze the cellbiological features. METHODS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by improved tissue cultivation. Immunophenotype and cellcycle were analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated by in vitro osteogenic and adipogenic induction as wel . RESULTS AND CONCLUSION:Some fusiform cells crawled out from human umbilical cord tissues after cultivation for 5 days and formed colonies about 10 days later. When the removed tissues were further cultured, more cells appeared again within 2 days and formed colonies after 5 days. The isolated cells exhibited similar morphology of fibroblast-like shape after passage. Furthermore, the cells expressed CD90, CD105, but were negative for the markers of CD34, CD45, HLA-DR. Population doubling time of the cells calculated from the result of MTT was about 50 hours and cellcycle analysis showed that 41.24%cells were in the G 2/S phrase. Therefore, the isolated cells had a high prolification ability. In addition, the isolated cells could be induced into osteoblasts and adipocytes in vitro. In a word, the results of this study demonstrated that the cells from the second tissues culture possessed the biological characteristics of mesenchymal stem cells and more primary umbilical cord mesenchymal stem cells were acquired through the improved method.

BACKGROUND:Placenta is a valuable source of mesenchymal stem cels for stem cel therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cels. Here, we present a new method for isolating human placenta-derived mesenchymal stem cels suitable for banking strategies and for future clinical applications. OBJECTIVE:To analyze the biological characteristics of human placenta-derived mesenchymal stem cels cultured by tissue dissociating and colagenase digestion. METHODS: Human placenta-derived mesenchymal stem cels were obtained from human placenta by tissue dissociating and colagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated byin vitro adipogenic, osteogenic and chondrogenic induction as wel. RESULTS AND CONCLUSION:Human placenta-derived mesenchymal stem cels could be passaged stablyin vitro. Furthermore, the cels expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cels proliferated actively and began to grow logarithmicaly at days 3-5 folowed by a plateau period at day 6. In addition, the isolated cels could be induced into adipocytes, osteocytes, chondrocytesin vitro. In a word, the results of this study demonstrated that the tissue dissociating and colagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cels that can be stably cultured in vitro and have strong proliferative ability.

AIM:To investigate the expression of extracellular matrix metalloproteinase inducer(EMMPRIN)and hepatocyte growth factor(HGF)in non-small-cell lung carcinoma(NSCLC)and their relationship with lymphoid metastasis and prognosis.METHODS:Expression of EMMPRIN and HGF in 77 cases of patients with NSCLC was detected immunohistochemically.The relationship of expression of EMMPRIN and HGF with tumor size,smoking,histological type,differentiation,lymphoid metastasis,clinical stage,and prognosis was analyzed.RESULTS:The expressive rates of EMMPRIN and HGF were 68% and 44%,respectively.The expressions of EMMPRIN and HGF were associated positively with lymphoid metastasis(r=0.371 and 0.339,P0.05).The expression of EMMPRIN was associated with the expression of HGF in NSCLC.CONCLUSION:The expression of EMMPRIN and HGF is associated with lymphoid metastasis and prognosis in NSCLC.Overexpression of EMMPRIN and HGF implies infavourable prognosis in NSCLC.

Objective To study the feasibility and safety of performing nephrectomy together with the removal of complicated inferior vena cava tumor thrombus under profound hypothermia and arrested circulation. Methods After made the median thoraco-abdominal incision, the exploration of the abdominal organs was done. The right kidney, inferior vena cava and renal pedicle were well exposed then. After the whole body heparinization, cannulas were put into ascending aorta, superior vena cava, aortic root and right superior pulmonary vein. The body temperature was reduced to 20℃ with cardiopulmonary bypass unit and the extracorporeal circulation was stopped then. Cut open the inferior vena cava at vena renalis dextra ingress and the F16 urinary catheter was inserted into atrum dextra through inferior vena cava and inflated. The tumor thrombus was pulled out and the right kidney was removed. The inferior vena cava incision was sutured to close and the extracorporeal circulation was resumed and patient was re-warmed.Results The operation time was 330 min and the extracorporeal circulation time was 90 min, while the profound hypothermia with circulatory arrest time was 20 min. The estimated blood loss during operation was 400 ml and 6 unit red cells and 600 ml blood plasm were transfused. The patient was awaked 2.5 h after the operation, food intake resumed 4 days after operation and the patient was discharged on day 10 post-operatively. After 6 months'follow-up, there were no local recurrence and metastasis occurred. Conclusion The technique of profound hypothermia and circulation arrest could improve the safety and efficacy in the treatment of renal cell carcinoma with suprahepatic (level Ⅲ) caval tumor thrombus.