Objective To investigate the effect of growth differentiation factor 5(GDF-5)on expression of gap junctional protein,connexin 43,during ehondrogenic differentiation of bone marrow sternal cells(BMSCs)in vitro.Methods BMSCs were isolated from mouse bone marrow and cultured in vitro.The cells in passage 3 were chosen to be induced into chondrogenic differentiation.After induction for 72 hours,TypeⅡcollagen protein was examined by immunocytochemistry and the sulfate glycosaminoglycan was measured by Alcian blue staining.With induction for 24,48 and 72 hours,the proliferation effects of BMSCs were investigated by MTT assay;connexin 43 mRNA and protein were examined by RT-PCR,western blotting and immunocytochemistry respectively at different time points during induction.Results According to MTT assay,GDF-5 had no effect on the proliferation of BMSCs at different time points of induction;RT-PCR,western blotting and immunocytochemistry showed that GDF-5 could promote expressions of connexin 43 mRNA and protein at different times during induction.After 72 hours of induction,immunocytochemistry showed expression of TypeⅡcollagen protein,and AIcian blue staining of proteo- glycan revealed deposition of typical cartilage extracellular matrix.Conclusion GDF-5 can enhance chondrogenic differentiation of mouse BMSCs in vitro by up-regulating the expression of gap junctional protein,connexin 43.

BACKGROUND:Animal experimental studies have confirmed that cell transplantation,neurotrophic factor infusion or transplantation as well as other methods can alter the local environment of injured spinal cord and promote its partial function recovery. OBJECTIVE:This study aimed to assess the clinical efficacy of olfactory ensheathing cell transplantation for the treatment of the sequel of myelitis,and to explore whether it would promote the recovery of the spinal cord function. DESIGN:A non-randomized self-control study. SETTING:Ward of Second Department of Surgery of Taian Disabled Soldiers Hospital of Shandong Province. PARTICIPANTS:Thirty-two patients with obsolete myelitis,who come from all over China and suffered from disease for 0.5 to 7 years,admitted to our hospital between June 2004 and July 2007 were recruited in this study.The involved patients,including 21 males and 11 females,were aged 5-48 years.Their neurological functions were not obviously improved after various conventional treatments and limb function exercise.Meanwhile,various sensorimotors and autonomic nerve functional impairments were left.Among the patients,18 suffered from acute viral myelitis,8 from acute purulent myelitis and 6 from tuberculous myelitis.After onset,they were all given large doses of radiosonde, dexamethasone,anti-inflammatory and immunomodulatory drugs and various neurotrophic drugs.Twenty-six patients presented complete injury and six patients incomplete injury.Informed consent of treatment was obtained from each patient.The therapeutic protocol was approved by the Ethics Committee of the hospital.Embryonic olfactory bulbs were harvested from aborted embryo,which was donated voluntarily by the patients or their relatives. METHODS:Cells were isolated from embryonic olfactory bulbs,cultured and purified for 7 to 14 days,and finally they were digested into single-cell suspension.Under the surgical miscroscope,the cells were transplanted onto the regions which were above or below the spinal cord injury site.Two weeks to 2 months postoperatively,neurological function of spinal cord was assessed by using the American Spinal Injury Association(ASIA)Scoring Standard formulated in 2000,and was compared to pre-operation function. MAIN OUTCOME MEASURES:①Sensory function change.②Motor function change. RESULTS:Half a year to 2 years after olfactory ensheathing cell transplantation,the sensory and motor functions of 32 patients were all obviously improved(motor function:55.72?10.50 vs.51.53?13.41;light touch:69.53?11.68 vs. 63.06?15.98;pain sense:69.50?12.20 vs.64.03?15.0,all P

Objective To investigate the incidence and the risk factors of major adverse cardiac events of critical care patients after noncardiac surgery.To study the incidence of myocardial injury after noncardiac surgery (MINS).Methods A retrospective analysis of critical care patients (n =1 087) after noncardiac surgery from January 2012 to January 2013 in our hospital was carried out.The clinical data of the medical history, intraoperative conditions, postoperative conditions and cardiac troponin Ⅰ (cTNI) were collected.Major adverse cardiac events included unstable angina, non fatal myocardial infarction, severe arrhythmia, heart failure and cardiovascular death.Risk factors of major adverse cardiac events of critical care patients after noncardiac surgery were analyzed using Logistic regression.Results The 30 d non cardiac surgery incidence of major adverse cardiac events was (94/1 087) 8.6％, the incidence of MINS was (168/1 087) 15.5％, Logistic regression analysis showed that the risk factors of major adverse cardiac events after non cardiac surgery were age (OR 1.03,95％ CI 1.01-1.05, P =0.002), a history of chronic renal insufficiency(OR 3.12,95％ CI 1.44-6.74,P =0.004), rise of cTNI 24 h after operation (OR 2.04,95％ CI 1.16-3.58,P =0.014) and use of vasopressor drugs within 24 h after operation (OR 2.34,95％ CI 1.25-4.38,P =0.008).Conclusions The incidence of major adverse cardiac events and the MINS of critical care patients after noncardiac surgery is high.Old age, history of chronic renal insufficiency, rise of cTNI 24 h after operation and vasopressor drugs within 24 h after operation are the independent risk factors.

Background and purpose:As a tumor suppressor gene, ribosomal S6 kinase 4 (RSK4) plays important roles in inhibiting cell proliferation, migration and inducing cell apoptosis. However, the proteins interacting with RSK4 are still unknown. This study aimed to screen proteins interacting with RSK4 in breast cancer cell line MDA-MB-231 by lfag-tag affnity puriifcation and LC-MS/MS (liquid chromatography/mass spectrometry).Methods:The pcDNA3.1/EGFP-RSK4-Flag eukaryotic expression vector was constructed by inserting full lengthRSK4 gene into vector pcDNA3.1/EGFP-Flag. And then the recombinant plasmids were transferred into MDA-MB-231 cells. Real-timelfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were used to detect the expression of RSK4 in MDA-MB-231 cells. Affnity puriifcation and LC-MS/MS were applied to screen proteins interacting with RSK4, and the related action mechanism of RSK4 with its interacted proteins was detected based on bioinformatics gene ontology (GO) and ingenuity pathway analysis (IPA).Results:Twenty-four proteins, such as serine/threonine-pro-tein kinase 38 (STK38)/serine/threonine-protein kinase 38-like (STK38L), MOB kinase activator 2 (MOB2) and protein arginineN-methyltransferase 5 (PRMT5), were successfully identiifed by Flag-tag affnity puriifcation followed by LC-MS/MS analysis, which probably interacted with RSK4. Bioinformatics analysis of the identiifed proteins suggested the proteins interacting with RSK4 were involved in diverse biological pathways, such as apoptosis and cell migration. Conclusion:According to bioinformatics results of proteins interacting with RSK4 identiifed by affnity puriifcation and LC-MS/MS, biological networks of RSK4 are involved in apoptosis and migration in breast cancer cells.

OBJECTIVETo evaluate the effects of Qingre Liqi Granule (QLG) on clinical therapeutic efficacy, electrogastrogram (EGG) and gastric emptying in patients with functional dyspepsia (FD).

METHODSThirty-two FD patients of dyskinesis type enrolled were treated with QLG by oral taking for 6 days. Scoring on 8 kinds of symptoms, including abdominal distension, abdominal pain, morning gastric fullness, belching, regurgitation, nausea, vomiting, and anorexia, fasting EGG and the gastric emptying determination were performed using single photon emission computed tomography (SPECT) before and after treatment.

RESULTSThe total and individual scores of clinical symptoms, expect that of vomiting, significantly decreased after treatment (P < 0.05), and the percentage of patients with tachygastria and bradygastria significantly decreased (P<0.01) at the same time. EGG after treatment showed significantly elevated rates of normal slow wave dominant power, and nearly normalized dominant frequency. An increased gastric emptying rate at different phases after 75 min (P < 0.05), and significantly shortened gastric emptying half-time (P < 0.01) were shown meanwhile in gastric emptying detection. The improvement of symptom score and gastric emptying half-time showed significant positive linear correlation (r =0.8929, P < 0.01).

CONCLUSIONQLG can improve symptoms of FD patients by regulating the rhythm and power of gastric electro-wave, increasing gastric motility and enhancing gastric emptying function.

Adult ; Aged ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Dyspepsia ; drug therapy ; Electromyography ; drug effects ; Female ; Gastric Emptying ; drug effects ; Humans ; Male ; Middle Aged ; Myoelectric Complex, Migrating ; drug effects ; Phytotherapy ; Treatment Outcome ; Young Adult

Objective To investigate the changes and clinical significance of the levels of matrix metallopro-teinase -9(MMP -9)in the serum and bronchoalveolar lavage fluid (BALF)in children with asthma whose had differ-ent levels of 25 -hydroxyl -vitamin D3 [25 (OH)D3 ]in the serum.Methods Fifty children with asthma between January 201 3 and January 201 5 were enrolled as the asthma group,based on the disease severity,and the patients were divided into the moderate to severe group (37 cases)and the mild group (1 3 cases),while 20 children with abnormal airway or tracheal foreign body were as a control group.The levels of 25 (OH)D3 and MMP -9 in the serum,levels of MMP -9 in BALF were measured and compared by using enzyme linked immunosorbent assay (ELISA)me-thod.Re-sults The level of serum 25 (OH)D3 in the asthma group[(1 9.5 ±6.4)μg/L]was lower than that in the control group[(39.3 ±7.1 )μg/L ],and there was a statistical difference between 2 groups(P <0.05).The level of serum 25(OH)D3 in moderate to severe group[(1 6.6 ±4.1 )μg/L]was lower than that in the mild group[(27.9 ±4.5)μg/L],there were statistically significant difference between 2 groups(P <0.05).There was no significant difference in the total number of cells and the percentage of macrophages among different serum 25(OH)D3 levels of children with asthma and the control group(P >0.05).The percentages of neutrophils,eosinophils and epithelial cells in BALF were significantly higher in different serum 25(OH)D3 levels of children with asthma than those of the control group(all P <0.05).There were significant difference levels of MMP -9 in the serum and BALF among different serum 25(OH)D3 levels of children with asthma and the control group(all P <0.05).In children with asthma,the levels of 25(OH)D3 in serum were significantly negatively correlated with the percentages of neutrophils,eosinophils and epithelial cells in BALF (r =-0.683,-0.795,-0.670,all P <0.05 ),exiting a significantly negatively correlation also seen between the serum levels of 25(OH)D3 and MMP -9 (r =-0.796,P <0.05).Conclusions Children with asthma often have low levels of serum 25 -(OH)D3 ,25(OH)D3 and MMP -9 may be involved in airway inflammation and airway remo-deling in children with asthma,and they may involve in the occurrence and development of asthma.

Objective To construct a reverse genetic platform for influenza B virus and to rescue influenza B virus.Methods Eight plasmids carrying the gene segments of B/Florida/4/2006 virus were constructed by using the bidirectional promoter vector pHW2000.293T cells were co-cultured with MadinDarby canine kidney (MDCK) cells and then transfected with the eight plasmids.The supernatants of cell culture and cell debris were collected after transfection and then injected into embryonated chicken eggs and MDCK cells for rescuing the influenza B virus strains.Results This reverse genetic system could be used for the preparation of reassortant influenza B virus strains.The titers of hemagglutination units of the rescued virus achieved 128-256/50μl.Most of the reassortant virus particles were spherical under electron microscope.Conclusion The pHW2000 reverse genetic system could be used for the rescue of influenza B virus.Moreover,it could also be used for the construction of influenza B virus with specific mutations for further in vestigation on the characteristics of influenza B virus and the construction of vaccine strain.

pH, affects of different alkaline materials KOH, K 2CO 3, NaOH, Na 2CO 3 on the growth, and NaCl, KCl tolerance of 29 isolates from the saline and alkaline soils in Xinjiang and Qinghai Provinces of China and 1 type strain were studied. Results showed that only a few alkaliphilic actinomycetes were Na +-obligately dependent, and K +-sensitive. Some alkaliphilic actinomycetes were CO 3 2- -sensitive, and NaCl, KCl could inhibit their growth. 4 kinds of alkaline materials had no affect on growth of alkaliphilic Nocardiopsis, and these strains showed high tolerance to NaCl, KCl. So it was presumed that only K + and CO 3 2- obligately dependent alkaliphilic Actinomycetes maybe exist in alkaline environments.

Objective:To investigate the feasibility and safety of endovascular repair for traumatic thoracic aortic pseudoaneurysm.Methods:From Oct 2015 to Oct 2018, the clinical and followup data of 7 patients diagnosed as traumatic thoracic aortic pseudoaneurysm in Fuwai Hospital of Chinese Academy of Medical Sciences were analyzed retrospectively.Results:The patients average age was (51.2±11.0) years old. All patients underwent surgery in the hybrid operating room under general anesthesia. Two did thoracic endovascular aortic repair (TEVAR), three did TEVAR combined with chimney technique to reconstruct the left subclavian artery, and 1 had TEVAR combined with fenestration to reconstruct the left subclavian artery. One did TEVAR with left common carotid artery and left subclavian artery bypass. The mean operative time was (90.1±27.4) min, the mean postoperative hospital stay was (8.9±3.7) d, and the mean postoperative follow-up time was 42.4 months. All the patients received CTA reexamination of the aorta after 1, 6, 12 months and yearly thereafter. TypeⅠendoleak was found in one patient with chimney technique to reconstruct of left subclavian artery after operation. CT showed that the type Ⅰ endoleak disappeared 6 months after operation. There was no death, paraplegia or stroke during the perioperative period and follow-up period, and there was no aortic related reintervention.Conclusion:TEVAR is a safe and effective method for the treatment of traumatic pseudoaneurysm of thoracic aorta, and the early and mid-term results were satisfactory.

Objective:To evaluate the safety and feasibility of the in-situ needle fenestration combined with the in vitro physician modified fenestration technique to reconstruct supra-aortic branches during thoracic endovascular aortic repair (TEVAR) for aortic arch lesions requiring landing at Z0 and Z1.Methods:From Nov 2017 to Dec 2019, eighteen patients who underwent both the in-situ needle fenestration and the in vitro physician modified fenestration techniques to extend the proximal landing zone to Z0 and Z1 during TEVAR were included in our study.Results:Sixteen patients underwent in vitro physician modified fenestration ,two patients underwent in vitro physician modified fenestration to reconstruct both the left common carotid artery and the innominate artery. All eighteen patients received in-situ needle fenestration to preserve the left subclavian artery. Supra aortic branches were preserved in all patients (38/38, 100%). There was no Type Ⅰ endoleak. Type Ⅱ endoleak was found in four paitnets (4/18). Type Ⅲ endoleak occurred in one patient (1/18). Type Ⅳ endoleak in four patients (4/18). Type Ⅲ endoleak needed open aortic arch repair 6 months later. The median follow-up time was 12 months. One (1/18) died in 12 months and the other patients were doing well.Conclusions:The joint application of the in-situ needle fenestration and the in vitro physician modified fenestration to reconstruct supra-aortic branches during TEVAR for aortic arch pathologies requiring landing at Z0 and Z1 was satisfactory.