Volume 92  ·   Issue 24  ·  2012

Clinical research is a general term for a kind of scientific research carried out on the clinical work platform with patients as the main research objects in view of the problems encountered in clinical work. The biggest difference between clinical research and laboratory research is that the research conditions are limited by the clinical work platform. The clinical research plan should not only meet the needs of research and answering scientific questions, but also meet the requirements of clinical work, that is, it should be considered comprehensively under the conditions of ensuring medical quality and medical safety, conforming to ethics and feasibility, etc. The contradiction between scientificity and feasibility is prominent, and the design and implementation of research plan is difficult.

Objective To evaluate the clinical characteristics and survival factors of patients with duodenal gastrointestinal stromal tumors (GIST).Methods The clinical data of 41 patients with duodenal GIST were analyzed retrospectively at Cancer Hospital and Institute,Chinese Academy of Medical Sciences from June 1996 to August 2011.Kaplan-Meier method was used to calculate the recurrence-free survival rate and the Cox proportional hazard regression model employed for the recurrence-free survival analysis.Results The lesions of duodenal GIST were predominantly located in the descending ( n =26,63.4％ ) and transverse portions ( n =10,24.4％ ).Most duodenal GIST presented commonly with upper gastrointestinal bleeding (n =18,43.9％ ) and 12 cases ( 29.3％ ) were incidentally detected by physical examinations.Eight patients underwent pancreatoduodenectomy and 27 limited resection.The tumor size varied from 0.6 cm to 30.0 cm (mean:8.4 cm).The recurrence-free survival rates analyzed by Kaplan-Meier method at 1,2 and 5-year were 94.1％,77.5％ and 65.0％ respectively. The results of Cox proportional hazards regression model indicated that the patients with ＞ 10/50 HP mitotic count showed a worse recurrence-free survival than those with ≤ 10/50 HP ( HR =3.7,95％ CI 1.0 - 13.7,P =0.049 ).After adjusting other confounding factors,mitotic activity was one significant prognostic factor of recurrence ( P =0.024).There was no significant association between the risk of recurrence and other prognostic factors,including diagnostic age,tumor size,type of operation and the risk of aggressive behaviors (all P ＞ 0.05).Conclusions Mitotic activity is one prognostic factor of duodenal GIST.And R0 resection should be regarded as an optional treatment for duodenal GIST.

Objective To explore the serum level of 25(OH)D and its relevant influencing factors in patients on maintenance hemodialysis.Methods The serum level of 25 ( OH ) D was detected by electrochemiluminescence.A total of 162 patients on maintenance hemodialysis were divided into 3 groups according to the serum level of 25 ( OH ) D:vitamin D deficiency:25 ( OH ) D ≤ 15 μg/L,vitamin D insufficiency:25(OH)D≤30 μg/L and vitamin D normal:25(OH)D ＞30 μg/L.Age,gender,relevant biochemical indices,high sensitivity C-reactive protein (hsCRP) and HOMA-IR were compared among there groups.The associations between 25 ( OH ) D and the influencing factors of its levels were analyzed by Pearson's correlation.The independent factors correlated with the level of 25 (OH)D were estimated by multiple linear regression.Results The average level of 25 (OH) D was 10.3 ( range:4.0 - 43.5 ) μg/L and the prevalence of 25 (OH)D insufficiency and deficiency 88.9％ (144/162).Significant differences existed in age,cholesterol ( CHOL),low density lipoprotein ( LDL),lipoprotein (a) ( LP(a) ),creatinine (Cr),lg(hsCRP),lg( reductase inhibitor) ( lg(RI) ) and lg( vitamin D) ( lg(VitD) ) between three groups( all P ＜ 0.05 ).Pearson's correlation analyses revealed that the serum levels of albumin (ALB),Cr and Ca2+ were correlated positively with lg(VitD) while those of white blood cell,CHOL,LDL,Lp(a),age,RI and hsCRP were correlated negatively with VitD.Multiple linear regression analyses demonstrated that hsCRP,RI,LDL and age were the independent influencing factors of the serum levels of 25 (OH)D.Conclusions The prevalence of 25 ( OH ) D insufficiency and deficiency is high.And microinflammation and insulin resistance are closely correlated with the insufficiency and deficiency of 25 ( OH ) D. The independent influencing factors of serum 25 ( OH ) D are hsCRP,RI,LDL and age.

Objective To evaluate the incidence and risk factors of lower extremity amputation among inpatients with diabetic foot.Methods For this retrospective study,a total of 685 inpatients with diabetic foot (Wagner grade 1 -5 )admitted at a multi-disciplinary Diabetic Foot Care Center,West China Hospital,Sichuan University during January 1,2005 and June 30,2011.The data of each patient including clinical information,laboratory results and final outcome were collected and analyzed.They were divided into non-amputated and amputated groups.And the latter included minor and major amputation groups according to amputation site.Results The overall amputation rate was 11.4％ in diabetic foot inpatients. The incidences of minor amputation and major amputation were 5.4％ and 6.0％ respectively. 17.9％ of amputated patients experienced a second amputation.The amputated patients had a longer hospitalized stay,higher counts of white blood cells and neutrophils,greater HbA1c and lower serum levels of hemoglobin and albumin than the non-amputation patients (P ＜ 0.05 ).The prevalence of diabetic peripheral neuropathy was significantly higher in the amputation group than that in the non-amputation group (P ＜ 0.05 ).However,no difference existed between the minor and major amputation groups ( P ＞ 0.05 ).Ankle brachial index (ABI) in the amputation group was significantly lower than that in the non-amputation group ( 0.41 ± 0.25 vs 0.91 ±0.36,P ＜0.01 ).Minor and major amputation inpatients had similar ABI (0.43 ±0.24 vs 0.39 ± 0.26,P =0.087 ).Ordinal regression showed that HbAlc ( P =0.015 ),ABI ( P =0.016 ),history of amputation ( P ＜ 0.01 ) and Wagner grade of diabetic foot ( P ＜ 0.01 ) were the independent risk factors of amputation.Conclusions Diabetic foot inpatients have a higher rate of lower extremity amputation.The risk factors of amputation include HbA1 c,ABI,history of amputation and Wagner grade of diabetic foot And diabetic peripheral neuropathy,ischemia of lower limbs (especially peripheral arterial diseases below knees),infection and nutritional state are closely associated with amputation of diabetic foot inpatients.

Objective To evaluate the effect of pitavastatin on high sensitivity C-reactive protein (hsCRP) in patients with hypercholesterolemia,and determine risk factors for the effect.Methods This study was a 12-week,multicenter,open-label,without parallel-group comparison,phase Ⅳ clinical trail Results There were 330 subjects in the per protocol set.Contrast to the baseline,the average levels of hsCRP in all of subjects and the group without a history of receiving previous statin medication at week 12 post-treatment decreased respectively 26.4％ ( 1.20 mg/L vs 1.68 mg/L) and 27.5％ ( 1.21 mg/L vs 1.97 mg/L,all P ＜ 0.05 ).The results of multilevel models indicated that the average levels of hsCRP reduced with the passage of treatment time,the time-varying rate of per-visit was 0.97 mg/L (95％ confidence interval 0.96 - 0.98 ).Controlled individual background covariates,the model predicted that pulse pressure and white blood cell count on the baseline had the significant positive effects on hsCRP( P ＜0.01 ).Conclusions Pitavastatin decreases hsCRP in patients with hypercholesterolemia.The main risk factors for the effect are pulse pressure and white blood cell count on the baseline.

Objective To evaluate the efficacy and safety profile and to explore the role of docetaxel,S-1 plus cisplatin (DCS) or oxaliplatin (DOS) in the treatment of advanced gastric cancer.Methods A total of 45 patients with advanced gastric cancer were recruited.They received DCS or DOS at the discretion of investigators.Docetaxel was given intravenously at the dose of 60 mg/m2 at d1,S-1 60 mg· m-2 ·d-1 or 80 - 120 mg/d according to individual patient's area of body surface orally from d1 to d14 and cisplatin 30 mg/m2 at d1,d2 or oxaliplatin 111 - 127 ( median:117) mg/m2 at d2.Each cycle was for 21 days.Results Forty-three patients received ≥ 1 complete cycle of DCS/DOS with a median cycle number of 5 ( range:1 - 8 ).Among 42 patients evaluated for efficacy,the outcomes were partial response (n =28),stable disease(n =9) and progression(n =5).The response rate was 66.7％.Progression-free survival(PFS) of 32 patients on chemotherapy alone was 7.1 months and the median overall survival (OS) was not reached. The most common grade 3/4 adverse effects included neutropenia (46.5％),thrombocytopenia ( 9.3％ ),vomiting ( 9.3％ ),nausea ( 7.0％ ) and diarrhea ( 4.7％ ).Ten of fourteen patients with advanced unresectable gastric cancer without clinically detectable distant metastases underwent surgical resection after a median of 4 ( 2 - 6 ) cycles of DCS or DOS and 9 ( 64.3％ ) had R0 resection.Conclusions DCS/DOS is effective for advanced gastric cancer and in the setting of neoadjuvant chemotherapy.And the toxicities of DCS/DOS are manageable.

Objective To explore the clinical features,diagnosis and management of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) in children.Methods The clinical manifestations,examinations,diagnostic modalities and treatments of children identified as ALCAPA at Beijing Anzhen Hospital from September 1993 to September 2011 were retrospectively reviewed.Results A total of 27 children were recruited.There were 16 males and 11 females with an age range of 1 month to 13 years.Among them,19 patients had an onset age of ≤1 years.The initial symptoms were acute heart failure ( n =15),pneumonia ( n =7 ) and cardiac dilation with murmur ( n =5 ).The pre-admission misdiagnoses included endocardial fibroelastosis (EFE) ( n =13 ),dilated cardiomyopathy ( DCM ) ( n =4 ) and mitral severe regurgitation ( n =2 ).The definite diagnosis was established on the basis of electrocardiography (ECG),echocardiography and 64-slice computed tomography or angiography.Twenty-six cases underwent immediate surgery after an accurate diagnosis,including left coronary artery (LCA) ligation ( n =1 ),LCA ligation plus coronary artery bypass grafting ( n =1 ),Takeuchi operation ( n =7 ) and LCA reimplantation (n =17).Six died postoperatively.And 20 cases were routinely followed up with symptoms,signs,ECG and echocardiography for 3 - 192 months.The outpatient visits were at Months 3,6 and 12 post-operation and then annual check-ups by returning to hospital,telephone,letters and other forms.The cohort had no overt symptoms and left ventricular ejection fraction returned to normal range.No complications occurred after LCA reimplantation.Conclusions As a rare congenital heart anomaly,ALCAPA may be definitely diagnosed with clinical features and serial diagnostic methods.It can be treated with several types of cardiac surgery.Coronary reimplantation is the technique of choice for surgical correction of ALCAPA due to its excellent prognosis.

Objective To explore the effects of canonical Wnt signaling transduction pathway on the proliferation and secretion of fibroblasts of uterosacral ligament so as to elucidate the pathogenic mechanism of pelvic organ prolapse (POP).Methods Five patients with grade ≥ Ⅲ POP and five with other benign gynecologic disorders as control were recruited. Specimens were taken from uterosacral ligaments and fibroblasts were cultured and purified.After the confirmation of cultured fibroblasts by immunohistochemical staining,their growth and proliferation activity were detected by thiazolyl blue tetrazolium bromide (MTT) assay and the expressions of matrix metal proteinase-2 (MMP-2),Wnt16,FZD5 and β-catenin by Western blot.Results ( 1 ) Over 90％ of the cultured cells were fibroblasts of vimentin positive and keratin negative.(2) The growth and proliferation activity of fibroblasts were significantly lower in the POP group than those in the control group ( P ＜ 0.01 ).(3) The levels of MMP-2 and FZD5 in the POP group were significantly higher than those in the control group (P ＜0.05 and P ＜ 0.01 respectively) while those of Wnt16 and β-catenin were significantly lower than the control group ( both P ＜ 0.05 ).Conclusions Canonical Wnt16 signaling transduction pathway becomes inhibited in the fibroblasts of POP patients resulting in lower growth and proliferation activity and reduced secretion of collagen. Elevated MMP-2 contributes to the increased degradation of collagen.And MMP-2 and Wnt16 signaling transduction pathway may be important factors for the changes of collagen state in pelvic organ support structures leading to POP.

Objective To explore the regulative factors on CD8 + HLA-DR+ T cells in the patients with severe aplastic anemia (SAA) and examine the roles of these cells in the immunopathogenesis of SAA.Methods CD8+ HLA-DR+ T cells were sorted from bone marrow mononuclear cells of 13 SAA patients from July 2011 to March 2012 by magnetic activated cell sorting system and were divided into 3 groups:interleukin 2 (IL-2) group (0,0.1,1,10,100 and 1000 U/ml),cyclospofine A (CsA) group (addition of 400 ng/ml CsA in each IL-2-containing well ),receptor antagonist group (addition of IL-2 receptor antagonist 8 μg/ml in each IL-2-containing well).Then cell proliferation rate was evaluated by MTT assay after a 72-hour culturing.Bone marrow mononuclear cells of the SAA patients were divided into CsA group,IL-2 group and control group and cultured for 18 hours and another 4 hours following the dosing of phorbol ester.The expression of tumor necrosis factor β (TNF-β) in CD8 + HLA-DR+ T cells was analyzed by flow cytometry.Results The cell proliferations of IL-2 wells at the concentrations of 10,100 and 1000 U/L (0.36 ±0.12,0.41 ±0.12,0.46 ±0.14) were significantly higher than those of the control wells (0.23 ± 0.11),CsA group (0.18 ±0.05,0.19 ±0.00,0.20 ±0.04) and receptor antagonist group (0.18 ±0.05,0.17 ± 0.04,0.18 ± 0.03,all P ＜ 0.05 ).No statistic difference existed between CsA and receptor antagonist groups ( P ＞ 0.05 ).The expressions of TNF-β of CD8 + HLA-DR + T cells of the IL-2 group were higher than those of the control group (64％ ±25％ vs46％ ±22％ ) whereas the CsA group (27％ ±20％ ) were lower than those of the control group ( both P ＜ 0.05 ),Conclusions IL-2 can significantly stimulate the proliferation of CD8 + HLA-DR + T cells and accelerate the in vitro secretion of TNF-β in SAA patients.The proliferation may be inhibited by CsA and receptor antagonist. And the expression of TNF-β is suppressed significantly by CsA.

Objective To retrospectively analyze the curative efficacy of umbilical cord blood transplantation (UCBT) with improved myeloablative conditioning regimen (total body irradiation (TBI)/cytosine arabinoside ( Ara-c)/cyclophosphamide (CY) without antithymocyte globulin ( ATG ) ) in adult patients with hematological malignancies. Methods Forty consecutive adult patients with hematological malignancies received improved myeloablative unrelated CBT at a single center from September 2006 to May 2011.Their average age was ( 23 ± 6 ) years and the average weight ( 58 ± 9 ) kg.Thirty-five ( 87.5％ )patients were high-risk and 15 (37.5％ )at the advanced disease status at pre-transplantation.They received 1 ( n =23 ) or 2 ( n =17 ) cord blood units.Seventy-five percent of them were transplanted with 1/2- human leukocyte antigen mismatched unit.The conditioning regimen consisted of 12 Gy TBI,granulocyte colonystimulating factor (G-CSF) plus Ara-c and CY without ATG. All patients received a combination of cyclosporine (CsA) and mycophenolate mofetil (MMF) for the prophylaxis of graft-versus-host disease (GVHD).Results For the entire group of patients,the average cell doses infused were (4.1 ± 1.1 ) × 107 total nucleated cells/kg and ( 2.4 ± 1.0) × 105 CD34+ cells/kg.All patients acquired engraftment with an implantation rate of 100％.The average time of absolute neutrophil count (ANC) ≥ 0.5 × 109/L was (20 ±5) days and the average time of platelet ≥20 × 109/L was(38 ± 12)days.Acute GVHD occurred in 23 patients( 57.5％ ) and 4 ( 10.0％ ) were of grade Ⅲ - Ⅳ.Chronic GVHD occurred in 22.9％ ( 8/35 ) evaluable patients.Relapse occurred in 12.5％ (5/40) patients.During a median follow-up period of 19.8 ( range 4.6 - 55.0 ) months,the transplantation-related mortality was 15.0％ (6/40) within 100 days and 35.0％ (14/40) within 1 year.The main causes of mortality were pneumonia and severe acute GVHD.Two-year overall survival (05) or disease-free survival was 58.8％ and 58.8％,respectively.Two-year OS for patients with advanced or complete remission disease was 48.6％ and 63.8％,respectively.Conclnsions The TBI/Ara-c/CY myeloablative conditioning regimen is well-tolerated and capable of establishing sustained donor cell engraftment and decreasing the risks of transplant-related death in adults with hematologic malignancies.For the high-risk and advanced patients,it may reduce the occurrences of relapse and chronic GVHD.

Objective To explore the effects of hyaluronic acid (HA) on biomechanical properties for porcine acellular dermal matrix (PADM) plus thin skin autograft after transplantation.Methods The dorsa of 10 Japanese white rabbits were symmetrically divided into four areas of A-D by random grouping.Full-thickness skin defects were created in Groups A-C while Group D was blank with normal skin.Operations were performed in Group A:implant with HA + PADM + thin skin autografts,Group B:implant with PADM + thin skin autografts and Group C:skin autografts group.Histological examination of specimen was performed at Day 56 postoperatively.And the biomechanical properties such as relaxation and stress-strain properties of grafts were recorded.Results The structure of PADM was found to be basically intact by hematoxylin and eosin E dyeing in Groups A and B.In Group A,dense fiber structure could be observed.Lots of regularly arranged collagenous fibers and new blood capillaries were grown into the dermal matrix with sparsely distributed inflammatory cells.In Group B,acellular dermal matrix became clustered with a small amount of invaded fibroblasts.And there was a high expression of inflammatory cells.The biomechanic pedormances of transplanted skin were:Group A's curve was mostly close to that of Group D's,Group B's curve was the most further from that of Group D's (P =0.001 ) and Group G's curve stayed between Groups A and B.Under the same strain,the stress of Groups A-D was ( 87 ± 8 ),( 115 ± 9 ),(60 ± 7 ) and (81 ± 4) kPa respectively.No significant difference of stress existed between these two groups (P=0.838).There was significant difference of stress between Groups B/C and D (P =0.001 and P =0.009).Conclusion Topical hyaluronic acid may be used to enhance the biomechanics pedormances of transplanted skin.

Objective To review the experiences and key factors of establishing an abdominal cardiac transplantation model in rats at National Institutes of Health Clinical Center.Methods Dark Agouti rats served as donor while Brown Norway rats as recipient.Cardiac transplantations were performed with a modified version of heterotopic cardiac transplantation.Proficiency was defined as 20 consecutive operations with ＜ 15％ mortality and no significant major co-morbidities.Results We performed 10 donor harvestalone operations and 70 transplantations to reach the proficiency level The survival rate of the last 20 consecutive surgeries was 95％.The mean total operative duration was 55 ruin and mean ischemic time of donor hearts 35 min.Conclusion To ensure a consistently high operative success rate,a surgeon must not only be proficient in micro-surgical skills,but also closely follow the standardized training procedures and strictly observe the animal care and use guidelines.

Objective To investigate the effect and potential mechanism of atorvastatin against H2O2-induced apoptosis of human umbilical vein endothelial ceils (ECV-304).Methods ECV-304 cells were pretreated with different concentrations of atorvastatin (0.1,1 and 10 μmol/L) for2 h,followed by an exposure to 100 μ mol/L H2O2 for 18 h.Cellular morphology was observed under fluorescence microscope.Cellular viability and apoptosis were evaluated by methyl thiazolyl tetrazolium (MTT) and flow cytometry.Finally the expressions of cleaved caspase-3 and caspase-9 were measured by Western blot.Results H2O2 treatment caused an obvious apoptosis of ECV-304 cells and significantly decreased the cellular viability as characterized by a high percentage (50.71％ ) of apoptotic cells.Atorvastatin pretreatment inhibit cellular apoptosis induced by H2O2 (39.45％,20.53％ and 7.83％ ).Western blot assay showed that H2O2 treatment caused a high expression of cleaved caspase-3 and caspase-9 while atorvastatin pretreatment obviously inhibited the expression in a dose-dependent manner. Conclusions Atorvastatin inhibits the H2O2-induced apoptosis of ECV-304 cells in a dose-dependent manner.This effect may be associated with the down-regulation of cleaved caspase-9/caspase-3.

Objective To explore the role of endoplasmic reticulum stress in brain injury following chronic intermittent hypoxia (CIH) in weanling rats.Methods A total of 48 male healthy Sprague-Dawley rats (3 - 4-week-old,80 - 100 g) were randomly divided into 4 groups:2-week-CIH (21H) group,4-week-CIH (4IH) group,2-week-control (2C) group and 4-week-control (4C) group.The morphologic changes were observed by hematoxylin-eosin (HE) staining and cell apoptosis detected by terminal deoxynuclectidyl transferase-mediated dUTP-biotin nick end labeling assay.Then hippocampus and prefrontal cortices were collected for transcription and expression analysis of glucose regulated protein 78 ( GRP78 ) by reverse transcription (RT) -PCR and Western blotting respectively.And the expressions of Caspase-12 mRNA and Caspase-12 protein in prefrontal cortexes were analyzed by RT-PCR and immunohistochemistry. Results The neuronal apoptosis in hippocampus and prefrontal cortices in CIH exposed groups were more pronounced than those of the control groups (all P ＜0.01),especially in the 4IH group (hippocampus:8.78％ ±0.71％ vs 3.26％ ± 0.45％,cortices:6.02％ ± 0.32％ vs 2.91％ ± 0.29％ ).The expression levels of GRP78 mRNA (hippocampus:0.424 ±0.033 vs 0.326 ±0.013 and 0.444 ± 0.028 vs 0.310 ±0.015,cortices:0.514±0.038 vs 0.430 ±0.017 and 0.524 ±0.038 vs 0.439 ±0.033) and GRP78 protein in hippocampus and prefrontal cortices (hippocarnpus:0.221 ±0.032 vs 0.178 ±0.014 and 0.241 ±0.019 vs 0.170 ± 0.013,cortices:0.307 ± 0.012 vs 0.226 ± 0.022 and 0.311 ± 0.023 vs 0.225 ± 0.025 ),and the expression levels of Caspase-12 mRNA (0.396 ± 0.004 vs 0.323 ± 0.014,0.417 ± 0.011 vs 0.313 ±0.011 ) and Caspase-12 protein (0.334 ± 0.035 vs 0.197 ± 0.023,0.368 ± 0.079 vs 0.215 ± 0.024) in prefrontal cortexes in the IH groups all were more than those in the 2C and 4C groups ( all P ＜ 0.05 ).Conclusions Chronic intermittent hypoxia can up-regulate the GRP78 transcription and expression in brain regions associated with learning and memory.This may induce the endoplasmic reticulum stress and activate the Caspase-12 mediated apoptosis signaling pathway.In the end,neuronal apoptosis occurs.All these factors may play an important role in the impairment of learning memory during the exposure of growing rats to chronic intermittent hypoxia.

Objective To explore the effects of lipopolysaccharide (LPS)-induced myeloid-derived suppressor cells (MDSCs) on the proliferation of spleen T lymphocytes.Methods BALB/c mice were randomly divided into two groups:LPS group and normal control group.They were injected intraperitoneally with LPS and normal saline solution respectively.MDSCs were separated with CD11b immunomagnetic beads from the spleen extract of mice. The morphological characteristics of MDCSs were observed by Wright-Giemsa staining and the characteristic molecules on cell surface identified by flow cytometry.And the effects of MDSCs on the in vitro proliferation of T cells were determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT).Results The proportion of MDSCs in the spleen of the LPS group was much more than that of the normal control group ( 27.4％ ± 6.6％ vs 5.1％ ± 3.8％ ; t =5.06,P =0.007 ).CD11b + Gr-1 +MDSCs could be separated by CD11b immunomagnetic beads from the spleen of mice injected with LPS at a high purity of 84.0％ +4.2％.MTT method showed that the proliferation of T cells decreased significantly after a co-cultivation with CD11 b+ M DSCs versus the control group.And it was positively correlated with the number of M DSCs ( F =46.26,P =0.000 ).Conclusions A high purity of LPS-induced myeloid-derived suppressor cells may be separated with CD11 h immunomagnetic beads.And it has dose-dependent inhibitory effects on the proliferation of the spleen T lymphocytes.

Objective To elucidate the effect of transforming growth factor-beta 1 (TGF-β1) on peritoneal fibrosis and the regulation of gastric cancer adhering to mesothelial cells. Methods The peritoneal mesothelial cell line of HMrSV5 was used to determine the role of TGF-β1 in the regulation of gastric cancer cell adhering to mesothelial cells.And the mRNA and protein expressions of collagen Ⅲ and fibronectin were detected by adhesion assay,Western blot,immunofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR).Results ( 1 ) The treatment of 5 ng/ml TGF-β1 could induce the expressions of collagen Ⅲ and fibronectin in mesothelial cells at 24,48 and 72 h ( P ＜ 0.01 ).(2) As compared with the controls,the percentages of adhered HGC-27 and HSC-39 gastric cancer cells significantly increased under the treatment of TGF-β1 for 24 and 72 h.The increased adhesion percentages of HGC-27 were 65％ ± 5 ％ and 124％ ± 11％ ( P ＜ 0.05 ) while those of HSC-39 85 ％ ± 9％ and 146％ ± 17％ respectively (P ＜ 0.05 ).( 3 ) Arginylglycyl-aspartic acid (RGD) ( knockdown of minimal sites for cell-binding domain of extracellular matrix) decreased the number of cancer cells adhering to mesothelial cells under the stimulation of TGF-β1.And the decreased adhesion percentage of HGC-27 was 65％ ± 8％ (P ＜ 0.05).Conclusions TGF-β1 significantly stimulates the expressions of collagen Ⅲ and fibronectin in mesothelial cells.And it is associated with the increased adhesion of gastric cancer cell and offers a favorable environment for the dissemination of gastric cancer.

A 52-year-old male was admitted to the hospital with fever accompanied by leukopenia and thrombocytopenia for 1 week. One week ago, he had sudden high fever, accompanied by low back pain, chills, congestion of bulbar conjunctiva, face and chest, and low urine output at one time. Blood routine in the local hospital showed leukopenia and thrombocytopenia, and cephalosporin antibiotics and ciprofloxacin intravenous drip for 4 days were ineffective. On the morning of August 28, 2011, he went to the emergency department of the First Affiliated Hospital of Dalian Medical University. Blood routine showed that leukocytes were 1.15×109/L, hemoglobin 137 g/L, platelets 38.2×109/L, urine protein positive (+ +), alanine aminotransferase 117 U/ml, aspartate aminotransferase 304 U/ml, and was admitted to the hematology department on the same afternoon. He was healthy in the past, lived in a rural area, and had no clear history of mosquito bites and rat contact.

Stem cells have been used in regenerative medicine since 1968 and have a history of more than 40 years. In recent years, the research of stem cells has developed rapidly, and it has been used to repair damaged tissues and organs, treat degenerative diseases, immune diseases and tumors, etc. [1-2]; However, there are still different understandings about the risks of using stem cells, especially the risks of secondary or transformation tumors, which are summarized as follows.

A 27-year-old male was admitted to the Department of Cardiovascular Medicine of the First Affiliated Hospital of China Medical University on June 25, 2009, mainly due to precordial pain for 2 months and aggravated for 5 days. The patient developed precordial pain at rest 2 months ago, which was dull and unbearable in nature, accompanied by soreness in both upper limbs, and the symptoms relieved spontaneously after about 20 minutes, and was treated with medication in a local hospital (details unknown). Five days ago, the patient felt that the symptoms had aggravated last time, and developed precordial discomfort with weakness in the left upper limb after slightly moving. It improved after self-taking "Xiaoxintong", so he came to our hospital for systematic diagnosis and treatment. He had no history of hypertension or diabetes in the past.