Objective To develop a knee joint impact device to produce animal model of subchondral bone micro-fracture in rabbits.Methods A knee joint impact device was designed. 12 New Zealand rabbits were divided into group A (n=6) and Group B (n=6). In the group A, 6 patellar-femoral joints of the 6 rabbits were impacted (1.74 kg from height of 30 cm). In the group B, 6 femoral condyles of the 6 rabbits were impacted (1.14 kg from height of 28 cm). All the rabbits were evaluated with X-ray and MRI post injury. The femora of all the rabbits were observed with HE staining. Results No fracture line was found in all the animals with X-rays. The MRI of the 6 knee joints in the group A showed low signal intensities on T1 weighted, T2 weighted and fat suppression images. Histopathology demonstrated integrated cartilage and sunchondral bone plate, micro-fractures of cancellous and bleeding in marrow. The MRI of the 5 knee joints in the group B showed low signal intensities on T1 weighted images and high signal intensities on T2 weighted and fat suppression images. Histopathology demonstrated integrated cartilage and sunchondral bone plate, micro-fractures of cancellous and bleeding in marrow. No disorder was found with MRI and histology in the contral sides. Conclusion The impact device of the knee joint can produce controllable and reproducible subchondral bone micro-fracture in rabbits.

BACKGROUND:To date, there are few domestic reports about the influence of bone marrow mesenchymal stem cel s on T cel s proliferation in patients with aplastic anemia. And no study addresses the topic that if bone marrow mesenchymal stem cel s achieve immune regulation in aplastic anemia patients through inhibiting T cel s proliferation.
OBJECTIVE:To explore the effects of human bone marrow mesenchymal stem cel s on T cel s immune regulation in patients with aplastic anemia.
METHODS:Human bone marrow mesenchymal stem cel s were isolated, cultured and subcultivated in vitro. The morphological appearance of bone marrow mesenchymal stem cel s was observed and surface markers were measured by flow cyometry. The bone marrow mesenchymal stem cel s were co-cultured with T cel s extracted from peripheral blood of healthy volunteers and aplastic anemia patients for 7 days. The expressions of interferon-γ, interleukin-4 and interleukin-10 in the supernatants were detected with enzyme linked immunosorbent assay.
RESULTS AND CONCLUSION:The levels of interleukin-2 and interferon-γin the supernatant of aplastic anemia patients were significantly higher (P<0.05), while levels of interleukin-4 and interleukin-10 were significantly lower than that in healthy controls (P<0.05). Bone marrow mesenchymal stem cel s suppressed the elevated levels of interleukin-2 and interferon-γ, and enhanced the decreased levels of interleukin-4 and interleukin-10, thus regulating the immune dysfunction of aplastic anemia patients.

Objective To evaluate the safety and efficacy of FW-Ⅱ axial blood pump short term assistance for treating acute heart failure.Methods We selected 5 patients who were difficult to remove cardiopulmonary bypass,and implanted FW-Ⅱ axial blood pump by left atrium-pump-femoral artery.Perioperative hemodynamics,circulating markers of myocardial injury,vW factor levels in plasma and white blood cell-platelet aggregates in different rotational speed were observed and analyzed.Results All patients were assisted for(24.0 ± 2.6) h by FW-Ⅱ axial blood pump with speed of 7 000-9 000 r/min,flow of 1.9-3.0 L/min,and no mechanical dysfunction.Compared with mean artery pressure(MAP),cardiac index(CI) and systemic vascular resistance index (SVRI) before implantation of FW-Ⅱ axial pump was (50.29 ± 6.98) mmHg (1 mmHg=0.133kPa),(1.70±0.23)L· min-1 · m-2 and (2 009.86 ±129.46) dyn· s · cm-5,MAP and CI significantly increased,while SVRI significantly reduced at 8 000 r/min after pump assistance [(65.43 ± 6.90) mmHg,(2.53 ± 0.27) L · min-1 · m-2,(1 578.14 ± 356.70) dyn · s · cm-5,P ＜ 0.01).CK-MB and cTnI levels reached significant decrease after 12 hours of pump assistance and maintained low level [(66 ± 11) IU/L and (8.4 ± 3.8) μg/L,P ＜ 0.01].Compared with 7 000 r/min,vW factor levels significantly increased at 8 500 r/min [(2.59 ± 0.57) U/L vs (1.26 ± 0.43) U/L,P ＜0.01].Platelet activation and white blood cell-platelet aggregation was the lowest at 8 000 r/min,and reached the most high level at 7 000 and 9 000 r/min [(15 ± 3) ％,(33 ± 3) ％ and (31 ± 5) ％)].Conclusion FW-Ⅱ axial flow pump can be effectively used for short-term treatment of acute ischemic left ventricular failure,8 000 r/min is optimum speed to balance hemodynamics and biocompatility.

BACKGROUND:Hematopoietic reconstruction of malignant tumor in hematopoietic system is related to disease itself,pretreatment program and therapeutic tool after transplantation;especially,mobilization.collection and cryopreservation of auto-peripheral blood stem cell play a key role in successful reconstruction of hematopoietic system after transplantation.OBJECTIVE:To investigate the reconstruction of hematopoietic system through mobilization, collection and cryopreservation of auto-peripheral blood stem cell in patients with malignant tumor and analyze the effective factors on quantity and quality of auto-peripheral blood stem cell.DESIGN:Case analysis based on malignant tumor in hematopoietic system.SETTING:Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA;Department of Blood,Zhujiang Hospital,Nanfang Medical University.PARTICIPANTS:A total of 18 patients with malignant tumor in hematopoietic system were selected from Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.Their ages ranged from 1 6to 56 years.Among them,2 patients had acute myelogenous leukemia(AML),1 acute lymphoblastic Ieukemia(ALL),2 lymphoblastic Ieukemia (LL),2 chronic granulocytic leukemia(CGL),4 multiple myeloma(MM),and 7 non.Hodgkin lymphoma.Granulocyte colony-stimulating factor(G-CSF)was made by Chugai Pharmaceutical Company Limited (batch number:N3G31).METHODS:①All patients were mobilized with associated chemotherapy+G-CSF.Associate chemotherapy:Patients with leukemia were given 2 g/m2 arabinosyl cytosine every 12 hours from lhe first to the third days and 200 mg/m2 etoposide or 50 mg/m2 fludarabine from the first to the fifth days. In addition patients with MM were treated with arabinosyl cytosine as the same way mentioned above and with 1 g/m2 cyclophosphamide from the first to the second days. And patients with lymphoma were given 2 g/m2 cyclophosphamide from the first to the second days. When numbers of leucocyte of all patients decreased below 1.0×109L-1 after chemotherapy.G-CSF started mobilization and the collection was stopped with 5μg/(kg·d)subcutaneous injection.②When numbers of leucocyte increased to (4.0-10.0)×109 L-1,hemopoietic stem cells of peripheral blood were collected till the amount of mononuclear cells≥4.0×108/kq or numbers of CD34+ cells≥2.0×108/kg.And then,the samples were dealt with cooling device.maintained in liquid nitrogen at-196℃ and defrosted in water bath at 37-40℃.③Focal sites of patients were pretreated with local irradiation with 200 cGy/time and 5 times/week for 4 successive weeks.The total dosage was 40 Gy.At 48 hours later,(55.3±28.7)mL hemopoietic stem cells of peripheral blood were transfused back. And the duration from transfusion to collection was about(56.5±22.3)days.300 μg/d G-CSF was subcutaneously injected into all patients at 1 day after transplantation and the reaction was stopped at the phase of neutrophil≥0.5×109L-1. Finally. Refusing-staining rate of trypan blue of peripheral blood stem cell, amount of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were detected before and after thaw.MAIN OUTCOME MEASURES:①Collection of auto-peripheral blood stem cell;②survival rate and related markers of auto-peripheral blood stem cell after cryopreservation;③hematopoietic reconstruction of auto-peripheral blood stem cell after transplantation.RESULTS:All 18 patients with malignant tumor in hematopoietic system were involved in the final analysis.The mean collection time of auto-peripheral blood stern cell was 12.6 days after chemotherapy.the collection times were 1.9.total number of leucocyte was(8.93±1.27)×1 0.L-1 on the first day,and collection rate of mononuclear cell was (138.33±28.61)%. ②Refusing-staining rate of trypan blue of auto-peripheral blood stem cell was similar before and after cryopreservation[(96.26±1.33)%, (92.75±2.04)%,P＞0.05].in addition,after cryopreservation,recovery rates of mononuclear cells,CD34+ cells and granulation-monophyly progenitor cell were(91.96±1.37)%, (85.94±0.64)%and (87.69±4.53)%,respectively.Collection rate of mononuclear cells,number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were lower in patients with myeloma than in those with leukemia and lymphoma (t=2.524-3.268.P＜0.05).③At 15 days after transplantation,15 patients had the neutrophil≥0.5× 109L-1;at 20 days after transplantation,blood platelet was≥20 × 100 L-1.granulation-monophyly progenitor cells[(18.67-26.82)× 105/kg] of 5 patients grew poorly if the course of chemotherapy was more than 10 times.Among them,3 patients had delayed hematopoietic reconstruction after transplantation of auto-peripheral blood stem cell.CONCLUSION:①High-dose chemotherapy combined with G-CSF can shorten collection time of peripheral blood stem cell and improve collection rate of mononuclear cells.②Increase of chemotherapy times before transplantation can affect quantity and quality of auto-peripheral blood stem cell and cause delayed hematopoietic reconstruction.

Objective To investigate the change of HR,RR and arterial blood gas in the treatment of BiPAP ventilation in patients with acute pulmonary.Methods Fifty eight patients with acute pulmonary edema were randomized into two groups.The control group(n =29)were given conventional general treatment only,but treatment group(n =29)were given BiPAP ventilation besides conventional treatment.4 h later,heart rate (HR),respiratory rate(RR),SaO2,pH,PaO2 and PaCO2 were compared between the two groups.Hospitalization duration and incidence of invasive mechanical ventilation were recorded after discharge.Results Compared with pre-treatment,HR,RR,SaO2 and PaO2 in treatment group were improved significantly(HR 124 ± 12 beat/min vs 83 ±6 beat/min,t =5.372,P ＜0.01)(RR 37 ±5 beat/min vs 19 ± 8 beat/min,t =4.285,P ＜0.01)(SaO2 81.4％ ±5.4％ vs94.1％ ±4.2％,t=2.731,P＜0.05)(PaO2 53.2±5.4 mm Hg vs 89.1 ±8.5 mm Hg,t=5.763,P ＜0.O1).And these four indicators were also improved in control group after treatment,(HR 123 ± 10 beat/min vs 95 ± 8 beat/min,t =t =3.459,P ＜ 0.01)(RR 36 ± 7 beat/min vs 24 ± 6 beat/min,t =3.127,P ＜0.01)(SaO2 81.8％ ±5.7％ vs 88.3 ±4.5％％,t =2.314,P ＜0.05)(PaO2 53.5 ±4.6 mm Hg vs 72.8 ±9.5 mm Hg,t =3.756,P ＜0.01).HR,RR,SaO2 and PaO2 in treatment group were more significantly improved than that of control group(P ＜ 0.01 or P ＜ 0.05).Hospitalization duration in treatment group was significantly shorter than that of control group(9 d vs 15 d,t =3.763,P ＜ 0.01).The incidence of invasive ventilation were lower than that of control group too(but P ＞ 0.05.Conclusion These results suggested that BiPAP ventilation can regulate HR RR and blood gas value to accetable levels,shorten hoptipitalization duration and reduce the incidence of invasive ventilation.It is proved to be an effective therapeutic technique in the treatment of acute pulmonary edema patients.

Objective To study the impact of the off-pump coronary artery bypass compared to the on-pump coronary artery bypass technique on the rate of acute kidney injury.Methods A retrospective analysis of 824 patients accepted coronary artery bypass surgery in FuWai Hospital Surgical ICU from March 2011 to January 2013.Patients were divided into two groups according to the surgical approach,597 patients in on-pump group and 227 patients in off-pump group.Then compared the difference of acute kidney injury incidence in the two groups.Results 92 patients in the on-pump group occurred AKI,the incidence of AKI was 15.4％.32 patients in the off-pump group occurred AKI,the incidence of AKI was 14.1％.Two groups of patients had no statistical difference in the incidence of AKI,x2 =0.222,P =0.638.Based on the preoperative baseline creatinine clearance levels.We defined normal creatinine clearance (SCR ≥ 60 ml/min),mild renal insufficiency (45 ml/min ＜ SCR ＜60 ml/min),moderate renal dysfunction group(31 ml/min ＜ SCR ＜45 ml/min).Decline with the preoperative creatinine clearance rate,the AKI incidence was elevated significantly,P ＜0.05.Patients with moderate renal insufficiency,offpump patients had lower incidence of AKI than those in on-pump group,P ＜ 0.05.The incidence of renal replacement in the two group patients had no statistical differences.Conclusion The incidence of AKI after coronary artery bypass surgery had no statistical differences with or without pump.In the patients with preoperative moderate renal insufficiency,off-pump can reduce the incidence of AKI,but could not change the kidney the incidence of renal replacement treatment.

Objective To investigate the changes of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF levels and their correlation with clinical symptoms of schizophrenia inpatiens and their value in the auxiliary diagnosis of schizophrenia .Methods The case-control study was used .The levels of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF were measured by using the enzyme linked immunosorbent assay(ELISA) in 85 inpatients with schizophrenia and 85 healthy controls .Their changes in the case group were compared between before treatment and after 3-month treatment .The Pearson correlation analysis was used to analyze the correlation between the lev-els of IL-1β,IL-6 ,TNF-α,NGF and BDNF with the positive and negative syndrome scale (PANSS) score and the auxiliary diagnosis value of serum cytokine and neurotrophic factor levels were evaluated with the receiver operating characteristic (ROC) curve .Re-sults The levels of serum IL-1β(t=4 .560) ,IL-6(t= 4 .957) and TNF-α(t= 4 .799) before treatment in the schizophrenia case group were significantly higher than those in control group ,while the NGF(t= -4 .806) and BDNF(t= -4 .881) levels were sig-nificantly lower than those in the control group ,the differences were statistically significant (P<0 .01) .After 3-month treatment , the levels of serum IL-1β(t=4 .543) ,IL-6(t=4 .327) and TNF-α(t=4 .654) in the schizophrenia case group were significantly de-creased compared with before treatment ,while the NGF(t= -4 .641) and BDNF(t= -4 .876) levels were significantly increased , the differences were statistically significant (P< 0 .01) .IL-1β was positively correlated with the positive symptoms scores (r=0 .325 ,P<0 .01) ,IL-6 was positively correlated with the negative symptoms scores (r=0 .319 ,P<0 .01) ,TNF-α was positively correlated with the positive symptoms scores (r= 0 .281 ,P< 0 .01) ,NGF was negatively correlated with the positive symptoms scores(r= -0 .229 ,P<0 .05) ,BDNF was negatively correlated with the positive symptoms scores (r= -0 .272 ,P< 0 .05) .The cut-off values of serum IL-1β,IL-6 ,TNF-α,NGF and BDNF in the auxiliary diagnosis of schizophrenia were 40 .083 ,20 .037 ,17 .115 ,19 .998 ,584 .157pg/mL respectively ,the corresponding areas under the ROC were 0 .723 ,0 .772 ,0 .686 ,0 .712 and 0 .708 respectively ,the sensitivities were 0 .565 ,0 .871 ,0 .894 ,0 .859 and 0 .729 respectively ,and the specificities were 0 .871 ,0 .565 ,0 . 365 ,0 .494 and 0 .624 respectively .Conclusion The levels of serum IL-1 ,IL-6 ,TNF-α,NGF and BDNF have the correlation with the clinical symptoms of schizophrenic inpatients and have a certain value in the auxiliary diagnosis of schizophrenia .

It is one of the most important forms of nucleic acid methylation in gene epigenetic modification and the most common driving molecular event in tumors. Detection of the free form nucleic acid can not only assist in early diagnosis and prognosis of tumors, but also avoid the problems of invasion and tumor heterogeneity caused by traditional tumor biopsy methods. In the present paper, the application status of free nucleic acid methylation detection in tumor diagnosis was reviewed, the challenges faced by free nucleic acid methylation detection were analyzed, and prospect of its clinical application was prospected.

Objective: To isolate and identify exosomes from human serum, explore the feasibility of exosomal CEA for the diagnosis of colorectal cancer. Methods: Retrospective study.64 cases with colorectal cancer patients(41 cases with normal CEA results and 23cases with high CEA results), 20 cases with benign colorectal diseases patients and 40 cases with healthy people of department of physical examination from October 2015 to December 2016 in Tongji Hospital of Tongji University. Exosomes were isolated from these serum using ExoQuick, and then identified by using transmission electron microscopy, and Western Blot for morphology and molecular phenotype.The serum level of CEA and exosomal CEA was measureed by enzyme linked immunosorbent assay (ELISA). The diagnostic efficacy of serum Exosomal CEA concentration in the colorectal cancer by using U test and the subject work characteristic curve (ROC). Results: Exosomes in human serum was successfully extracted with ExoQuick kit.Exosomal CEA concentration in 64 cases with colorectal cancer patients (1 056.36-28 637.78)pg/ml was significantly higher than in cases with benign colorectal diseases patients (394.61-2 437.83)pg/ml and healthy controls(610.89-2 076.70)pg/ml(U=124.000, U=119.000, P<0.01). Exosomal CEA concentration in 41 colorectal cancer patients with normal serum CEA concentration(1 056.36-5 832.07)pg/ml was significantly higher than in benign colorectal diseases patients and healthy controls(U=113.000, U=119.000; P<0.01). ROC analysis of exosomal CEA yielded an AUC(area under the ROC curve)of 0.954(95% CI=0.919-0.988), which was higher than the serum CEA.The area under the ROC curve of serum Exosomal CEA concentration for colorectal cancer diagnosis is 0.954(95% CI=0.919-0.988), which is superior to the serum CEA concentration[0.636 (95% CI=0.531-0.742)]. Conclusions Isolation and detection of serum Exosomal CEA concentrations have a good diagnostic efficacy in colorectal cancer. It′s possible to be a marker for early diagnosis of colorectal cancer.(Chin J Lab Med, 2018, 41: 503-508)

Objective To explore the structure and component changes of colonization bacterium in colorectal cancer (CRC) and analyze the relationship between the colonization bacteria and the development of CRC.Methods Clinical data of this retrospective study were obtained from Tongji Hospital of Tongji University (1/2016-10/2017).Forty matching tissues,including normal intestinal mucosa tissues (n=12) and CRC tissues (n=28),were collected in the present study.The V3-V4 region of bacteria 16S rRNA gene was detected using Illumina Miseq sequencing technologies.The colonized bacterium structure and composition were analyzed and compared between the two groups.Results It was found that there is significant difference in structure of the colonized bacterium between the two groups.In the level of Phylum,the abundance of Firmicutes and Bacteroidetes were significant lower (Z=-1.964,P＜0.05),and Proteobacteria was significant higher (Z=-1.997,P＜0.001) in the tissues of CRC.In the level of genus,Bacteroides,Blautia,Prevotella,and Faecalibacterium were significant lower in the tissues of CRC (Z=-3.008,P＜0.05).Buchnera,Prevotella,and Prevotellaceae only existed in normal intestinal mucosa tissues and the abundance was top three.Catonella,Kurthia,and Brachymonas only existed in CRC tissues and the abundance was top three.Conclusions The overall structure and component were significant difference between the two groups.Some bacterium disappeared or reduced,and others new emerged or increased.The change of colonized bacterium structure and component in intestinal mucosa may play an important role in the development of CRC.Therefore,it may be an,accurate and low cost of early diagnosis marker by identifing the structure and component change of intestinal mucosa colonized bacterium for CRC,which may find a new way for prevention and treatment of CRC.