Objective To investigate the predictive value of extra pulmonary multiple factors including creatine kinase-isoenzyme MB (CK-MB) for the prognosis of patients with acute paraquat poisoning.Methods A retrospectively analysis were conducted on 641 patients who were treated at the First Affiliated Hospital of Zhengzhou University due to oral paraquat poisoning from October 2002 to April 2017.The observation end point was that the patients died from paraquat poisoning within 3 months after admission or were still alive within 3 months after paraquat poisoning.The patients' data were retrieved,including general information,the dose of poison,urinary paraquat concentration,arterial blood gas analysis,alanine transaminase (ALT),total bilirubin (TBIL),uric acid (UA),aspartate transaminase (AST),creatine kinase (CK),CK-MB,B type natriuretic peptide (BNP),lactic dehydrogenase (LDH),high sensitivity troponin T (hsTnT),C-reaction protein (CRP) and procaicitonin (PCT).According to the patient's prognosis within 3 months,the patients were divided into a survival group and a non-survival group.The above indicators were compared between the two groups and the diagnostic value of CK-MB for acute paraquat poisoning was analyzed according to the receiver operating characteristics (ROC) curve.Collect the last arterial blood gas analysis,and laboratory test results were analyzed by binary logistic regression analysis to determine the risk factors for death in patients with acute paraquat poisoning.Results Among the 641 patients with acute paraquat poisoning,315 (49.1％) patients survived and 326 (50.9％) died.Compared with the survival group,patients in the non-survival groupthere were older,had a shorter hospital stay,and had a higher oral paraquat dose and urinary paraquat concentration;Lac,TBIL,UA,AST,CK,CK-MB,BNP,LDH,CRP and PCT were higher,while blood gas analysis index were lower in the non-survival group (P＜0.05).Binary logistic regression analysis showed that the dose of paraquat,CK-MB and AST were closely related to the prognosis of patients with acute paraquat poisoning.The optimal cut-off value of ingestion dose,the first urinary paraquat concentration on admission and CK-MB in predicting the prognosis of patients with acute paraquat poisoning were 7 g (AUC=0.918,sensitivity 80.6％,specificity 87.5％,Yoden index 0.681,P＜0.01),5.16 μg/mL (AUC=0.879,sensitivity 93.8％,specificity 70.1％,Yoden index 0.639,P＜0.01),and 18.2 U/L (AUC=0.846,sensitivity 83.9％,specificity 71.9％,Yoden index 0.558,P＜0.01),respectively.Binary logistic regression analysis of the last biochemical indicators of paraquat poisoning showed that the dose of poison,the last CK-MB,the last SCr,urinary paraquat concentration,and the last blood Na+ were closely related to the prognosis of patients with acute paraquat poisoning.Among them,the last CK-MB＞18.05 U/L often indicated poor prognosis (AUC=0.808,sensitivity 79.7％,specificity 65.8％,Yoden index 0.455,P＜0.01).Conclusions In the treatment of patients with acute paraquat poisoning,there are significant differences in extra pulmonary factors such as heart,liver,kidney,electrolytes and inflammatory markers in patients with different prognosis,so the monitoring and follow-up should be improved,in addition to focusing on the presence and treatment of pulmonary fibrosis.In particular,CK-MB is an independent risk factor for the prognosis of acute paraquat poisoning.In the late stage of poisoning,CK-MB,SCr,and blood Na+ have a strong predictive value for the prognosis of the patients,and we should pay attention to the regular follow-up of the above mentioned laboratory items.

According to the pathological characteristics of symptomatic chronic thoracic and lumbar osteoporotic vertebral fracture (SCOVF), the different clinical treatment methods are selected, including vertebral augmentation, anterior-posterior fixation and fusion, posterior decompression fixation and fusion, and posterior correction osteotomy. However, there is still a lack of a unified understanding on how to choose appropriate treatment method for SCOVF. In order to reflect the new treatment concept and the evidence-based medicine progress of SCOVF in a timely manner and standardize its treatment, the clinical guideline for surgical treatment of SCOVF is formulated in compliance with the principle of scientificity, practicability and advancement and based on the level of evidence-based medicine.

In sonoporation, the cell membrane is broken-up temporarily by ultrasound mediated microbubbles, which is promoting drug or gene into the cell. In current literatures, there are numerous studies of single microbubble dynamics in sonoporation. However till now, little studies have been focused on the sonoporation incidence caused by more than one microbubble. In this article, the dynamic model of two adjacent microbubbles in stable cavitation has been introduced. By the model, the forces including secondary Bjerknes force on cell membrane given by microbubbles and their effects on sonoporation have been numerically studied. According to the experimental parameters, we numerically studied (1) effects of the ultrasound and microbubble parameters on the secondary Bjerknes forces; (2) the forces exerted on cell membrane by microbubble, including the secondary Bjerknes force; (3) the sonoporation possibility caused by those forces produced by microbubble. In this article, the ultrasound and microbubbles' parameters range were found to produce sonoporation by two adjacent microbubbles. Furthermore, it is the first time to found that the microbubbles' parameters are more important than ultrasound parameters on sonoporation.

Objective To investigate the effect of total alkaloids from Rhynchophylline on hypertensive target organs. Methods 40 spontaneous hypertensive rats (SHRs) were divided into 4 groups as follows: normal control group,positive control with Tianma Goutengyin granule at 750 mg?d-1 ,total alkaloids from Rhynchophylline at 2.5 mg?day-1 as low dosage,and at 15.0 mg?d-1 as high dosage.All the rats were fed up for 20 weeks,and baroreflex sensitivity (BRS,in ms?mmHg-1)was determined by revised Smyth’s method.Heart,brain and kidney of the rats were sectioned for histological analysis at the end of 20th week. Results In sixteenth weeks,BRS of positive control,Tianma Goutengyin granule group,total alkaloids at low dosage and high dosage were(0.27±0.05),(0.31±0.06),(0.35±0.08),(0.34±0.08) ms?mmHg-1,respectively.BRS in SHR was not decreased except in the positive control.The treatment groups were superior to the normal control group in structure of heart,but not in the brain and kidney of the rats. Conclusion Total alkaloids from Rhynchophyllinecould protect heart from hypertensive injury.

To improve higher training quality and capacity of orthopedic postgraduates,early contact education,problem-base learning combined with co-supervisor with radiologists were engaged for orthopedic postgraduates since 2008.Effect of the practice was assessed by postgraduates one year later.Investigation revealed that practice aroused learning enthusiasm and initiative,promoted comprehensive understanding of orthopedic knowledge,set up social responsibility,and constructed innovative thinking.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.

Objective To examine the gene expression profile of SRY-type high-mobility-group box-9(SOX9) during entochondrostosis of mice and investigate the effects of transfection of the pDC316-SOX-9 on mice mesenchymal stem cells(MSCs) in vitro. Methods cDNA microarray technique with 34 000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of SOX9 was performed with GCOS1.2 software. The recombined expression vector pDC316-SOX-9 was constructed and transfected into mice MSCs by lipofectamine. The phenotype changes of cells were observed with cell energometry, HE stain, immunohistochemical method, RT-PCR and ELISA.Results The gene expression of SOX9 during the critical phase of chondrogenesis in mice embryo limbs at E12 was increased evidently. SOX9 might promot chondrogenesis. As compared with vector and blank group,the chondrocytes of the SOX9 transfected group had the tendency of enhanced differentiation. Conclusion SOX9 may promote chondrogenesis. The transfection of SOX9 gene into mice MSCs can promot MSCs differentiate into chondrocyte, which may provide some experimental data for cartilage histoengineering.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: The bone marrow stromal cells in red bone marrow (RBM) are the non-specific bone growth factors,and the target cells of bone morphogenetic protein (BMP), and they have the activities of osteoinduction and osteogenesis.Injectable calcium alginate gel (CaAG) is a degradable biomaterial with good histocompatibility,it provides scaffold for the proliferation and adhesion of osteoblasts and chondroblasts,and it is good for the proliferation of new capillaries.OBJECTIVE:To prepare CaAG-BMP-RBM gel compound,and observe its osteoinductivity.DESIGN:A randomized controlled animal trial.SETTINGS:Department of Orthopaedics,Shenzhen People's Hospital;Department of Orthopaedics,Union Hospital affiliated to Tongji Medical College,Huazhong University of Science and Technology.MATERIALS:The experiments were carried out in Shenzhen People's Hospital from February 2002 to February 2003.Twenty-seven pure SD rats, either male or female,weighing (200±20) g,were purchased from the Animal Experimental Center of Southern Medical University (No.SCXK2002-99).BMP was bought from Xijing Hospital affiliated to the Fourth Military Medical University of Chinese PLA.METHODS:The rats were randomly divided into three groups:CaAG-BMP-RBM group(n=9),CaAG.BMP group (n=9) and CaAG group (n=9).In the CaAG-BMP-RBM group,the rats were anesthetized,then 0.1 mL RBM collected from trochanteric section of fetour was added into the prepared CaAG-BMP compound (0.4 mL),sufficiently mixed,and then injected into posterior femoral muscle.The rats in the CaAG-BMP group and CaAG group were implanted with CaAG-BMP and CaAG into bilateraI posterior femoral muscles respectively.①The conditions of becoming conscious after anesthesia, incision healing, diet and activities were observed, the size and consistency of the implants and distribution of vessels were also observed.②The indexes were detected at 1,2 and 4 weeks after model establishment respectively,and 3 rats were used for each time point.The sections were observed under light microscope.and the activity of alkaline phosphatase (ALP) was determined.MAIN OUTCOME MEASURES: ①General conditions of animals and gross observation of the implants;②Histopathological observation;③ALP activities.RESULTS: All the 27 SD rats were involved in the analvsis of results.The rats became conscious at 4.6 hours after anesthesia,and they could eat normally;Hematom at incision disappeared.and the rats could move normally at 24 hours;No sign of infection at incision was observed at 72 hours;The incisions healed completely and suture shed after 2 weeks.The incisions were stage Ⅰ healing in all the 27 rats.①Results of gross observation of the implants:At 1 week after implantation.the size of implant did not decrease and vessels in-grew in both the CaAG-BMP-RBM group and CaAG-BMP group;At 2 weeks,lhe quality was hard, and the section appeared as gray and white with deposition of bone-like tissues;At 4 weeks,a great amount of vessels in-grew,and there were many depositions of hard bone-like tissues.②Results of the histopathological observation under light microscope:In the CaAG-BMP.RBM group.many mesenchymal cells aggregated, osteoblasts and chondroblasts proliferated actively al 1 week;The chondrocytes tended to mature with cartilage-like and bone-like matrixes at 2 weeks; Many osteocytes were observed and fibrous bone formed at 4 weeks.which were more then those in the other two groups.③Results of ALP activity:At 1.2 and 4 weeks after implantation.the ALP activities in the CaAG group [(0.179±0.018),(0.058±0.017).(0.027±0.018)IU/g] were lower than those in the CaAG-BMP-RBM group[(0.922±0.226),(1.169±0.249),(0.431±0.081)IU/g,P＜0.01);At 1 and 4 weeks,the ALP activities in the CaAG-BMP group[(0.447±0.015),(0.276±0.081)IU/g]were lower than those in the CaAG-BMP-RBM group (P＜0.05).CONCLUSION:The compound of CaAG-BMP-RBM possesses stable and lasting osleoinductivity.

BACKGROUND: During recent years, mononucleotide polymorphism of some genes is possibly related to affectability of osteoarthritis (OA). However, previous researches mainly compare the gene expression of synoviocytes between OA and rheumatoid arthritis (RhA); therefore, the correlation of gene expression between synovioblast and fibroblast in other tissues should be further studied as compared with OA.OBJ ECTIVE: To observe the differences of gene expression between OA synovioblast and skin fibroblast.DESIGN: Observational contrast analysis.SETTING: People's Hospital of Shenzhen City.PARTICIPANTS: Synovium tissue was derived from OA patients who received replacement of knee joint in the Department of Orthopaedics, People's Hospital of Shenzhen City. All OA patients met the diagnostic criteria of osteoarthritis established by American College of Rheumatology in 1995. Three patients including 1 male and 2 females aged more than 65 years old and they did not have cardiac and pulmonary disease and diabetes mellitus. Three male normal volunteers who aged 25 to 35 years did not have rheumatic disease, osteoarthritis and dermatosis. All subjects provided a confirmed consent. The main reagents were detailed as follows: RPMI1640 culture medium, fetal bovine serum and TRIZOL agent (Invitrogen Life Technologies Company, USA); pGEM-T pUC (Progema Company, USA);Display PROFILE-BASIC and Display PROFILE Probe kits (Qbiogen Company, USA).METHODS: The experiment was carried out in People's Hospital of Shenzhen City from January to June 2005. Synovium of OA patients were treated with primary culture to obtain synovioblast; meanwhile, skin fibroblast treated with primary culture from normal subjects was regarded as the control group. Restricted enzyme section differential display was used to separate the different-expressed genes of synovioblast and skin fibroblast in OA patients. In addition, blast technique was used to compare the resulted ranks with Genbank ranks.MAIN OUTCOME MEASURES: Differences of gene expression between synovioblast and skin fibroblast in OA patients.RESULTS: Gene expressions of superoxide dismutase (SOD), TFPI2, CXCL2, CXCL6 and transforming growth factor (TGF) were high in synovioblast of OA patients as compared with those in skin fibroblast of normal subjects.CONCLUSION: Gene expressions of SOD, TFPI2, CXCL2, CXCL6 and TGF are high in synovioblast of OA patients as compared with those in skin fibroblast of normal subjects. This suggests that gene may play a certain role in onset of OA.