BACKGROUND:Fibrin glue introduced into calcium phosphate cement has not been confirmed whether this way could overcome the compressive limits and the low degradation of calcium phosphate cement and to modify the biological properties of calcium phosphate cement. OBJECTIVE:To explore the mechanical and biological properties of calcium phosphate cement/fibrin glue at different powder/liquid ratio for bone regeneration in vitro. METHODS:Calcium phosphate cement and fibrin glue were mixed at ratios of 1:1, 3:1, 5:1 (mL/g), and the pure calcium phosphate cement served as controls. Setting time, scanning electron microscope and the biomechanical test were used to analyze the composite scaffold structure, physical performance and the mechanical properties. Passage 3 osteoblasts were respectively inoculated on the material surface of the four groups, and pure cells served as blank controls. celladhesion, proliferation and alkaline phosphatase activity were observed. RESULTS AND CONCLUSION:The initial and final setting time of calcium phosphate cement/fibrin glue at 1:1 and 3:1 (mL/g) was higher than that in the control group (P<0.05), while the initial and final setting time of calcium phosphate cement/fibrin glue at 5:1 (mL/g) was lower than that of the control group (P<0.05). Scanning electron microscope showed smoother and denser surface of composite scaffolds compared with the pure calcium phosphate cement. The aperture of the composite scaffolds was decreased with the increasing concentration of fibrin glue. The compressive strength of composite scaffolds at 3:1 and 5:1 was higher than that of the control group (P<0.05), while the modulus of the composite scaffolds at 1:1, 3:1, 5:1 was higher than that of the control group (P<0.05). celladhesion, proliferation and alkaline phosphatase activity showed no difference among the three composite scaffold and control groups, but al higher than the blank control group (P<0.05). These findings indicate that fibrin glue introduced into calcium phosphate cement can overcome the low-strength limits of calcium phosphate cement, and maintain the good biological properties of calcium phosphate cement for bone regeneration.

Cardiac Catheterization ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Prosthesis Implantation ; instrumentation ; Septal Occluder Device

In order to separate and purify the PEGylated recombinant human granulocyte stimulating factor (rhG-CSF) at large laboratory-scale level, a two-step ion-exchange chromatographic separation procedure was designed. Cation-exchange chromatography was applied first to separate PEGylated rhG-CSF from un-reacted rhG-CSF, followed by anion-exchange chromatography to dissolve individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the free PEG. The molecular weight of individual PEGylated rhG-CSF was determined by MALDI-TOF and SDS-PAGE. MALDI-TOF mass spectrometry revealed that the molecular weights of mono-, di- and tri-PEGylated rhG-CSF are 23.8 kD, 28.6kD and 33.8kD, respectively. Cell proliferation activity was detected by MTT assay using NFS-60 cell. The in vitro residual bioactivity of mono-, di- and tri-PEGylated rhG-CSF were 90%, 75% and 43% respectively, comparing with the un-conjugated rhG-CSF. These results indicated that the un-conjugated rhG-CSF and excess free PEG can be removed completely and the three conjugate species can be purified into homogeneity by the two consecutive ion-exchange chromatographic steps. The purification procedure is easy to scale-up, high in performance and recovery.
Chromatography, Ion Exchange ; methods ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; chemistry ; isolation & purification ; Humans ; Polyethylene Glycols ; chemistry ; isolation & purification ; Recombinant Proteins

OBJECTIVEPPAR-gamma is associated with the differentiation, apoptosis, proliferation and cytokine secretion of immunologic cells. This study investigated peripheral blood lymphoblastic PPAR-gamma mRNA expression and its correlation with plasma IL-13 contents in children with acute idiopathic thrombocytopenic purpura (ITP).

METHODSFifty-three children with acute ITP who were in line with the standard test between September 2007 and July 2008 were enrolled. Fifty healthy children during the same period were used as the control group. PPAR-gamma mRNA expression in peripheral blood lymphocytes were detected by RT-PCR. Plasma IL-13 contents were detected using ELISA.

RESULTSPPAR-gamma mRNA expression in peripheral blood lymphocytes from acute ITP children were significantly higher than that in the control group (0.78 +/- 0.03 vs 0.52 +/- 0.05; P< 0.05). Plasma IL-13 contents in children with acute ITP were also significantly higher than those in the control group (160.21 +/- 34.26 pg/mL vs 121.42 +/- 12.69 pg/mL; P< 0.05). There was a positive correlation between plasma IL-13 level and lymphoblastic PPAR-gamma mRNA expression in children with ITP (r=0.89, P< 0.05).

CONCLUSIONSPPAR-gamma mRNA expression in peripheral blood lymphocytes increased and were positively correlated with plasma IL-13 contents in children with acute ITP, suggesting that PPAR-gamma and IL-13 might participate in the pathogenesis of acute ITP.

Acute Disease ; Child ; Child, Preschool ; Female ; Humans ; Interleukin-13 ; blood ; physiology ; Lymphocytes ; metabolism ; Male ; PPAR gamma ; genetics ; physiology ; Purpura, Thrombocytopenic, Idiopathic ; etiology ; immunology ; RNA, Messenger ; analysis

OBJECTIVETo explore the effect of Scalp acupuncture on serum neuron specific enolase (NSE) and S-100β concentrations, and incidence rates of postoperative delirium (POD) and postoperative cognitive function (POCD) of elderly patients undergoing hip replacement.

METHODSEighty-four patients undergoing scheduled hip replacement under combined spinal-epidural anesthesia (CSEA) were assigned to the control group (group C) and the scalp acupuncture group (group S) according to random digit table, 42 cases in each group. In group S, scalp acupuncture was additionally performed according to International Standardized Scheme for Scalp Acupuncture. Scalp acupuncture was performed during the operation from the MS1 middle line of forehead [1 cun before Shenting (GV24), including Shenting (GV24)] and MS5 middle line of vertex [from Baihui (DU20) to Qianding (DU21), including Baihui (DU20) and Qianding (DU21)]. The operation time and post-operative length of stay were observed. The midazolam dosage, hemorrhage amount, fluid transfusion amount, urine amount, use rates of ephedrine and atropine during the operation were also observed and compared between the two groups. The occurrence rate of POD and POCD at post-operative day 3 (T₁), week 1 (T₂), month 3 (T₃), and month 6 (T₄) were measured. Eighteen patients were randomly selected to collect blood from internal jugular vein before anesthesia t₀), immediately after ending the surgery (t₁), 6 h after operation (t₂), 24 h after operation (t₃), and 48 h after operation (t₄), respectively. Serum levels of NSE and S-100β were correspondingly measured.

RESULTSThere was no statistical difference in the operation time, midazolam dosage used during the operation, hemorrhage amount, fluid transfusion amount, urine amount, use rates of ephedrine and atropine (P > 0.05). Compared with group C, the post-operative length of stay was shortened in group S (P < 0.05). The incidence rate of POD and that of POCD at each time point were lower in group S (P < 0.05). The expression level of NSE decreased at t₂, t3, and t₄, and the expression level of S100β also decreased at t₁, t₂, t₃, and t₄(P < 0.05). There was no statistical difference in expression levels of NSE or S100β between the two groups at other time points (P > 0.05).

CONCLUSIONScalp acupuncture could attenuate central nervous system lesion and improve POCD of elderly patients undergoing hip replacement.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Arthroplasty, Replacement, Hip ; Cognition ; Humans ; Length of Stay ; Phosphopyruvate Hydratase ; blood ; Postoperative Complications ; Postoperative Period ; S100 Calcium Binding Protein beta Subunit ; blood ; Scalp

OBJECTIVETo determine whether U50488H, a selective agonist of kappa-opioid receptor, could induce biphasic (early and late) cardioprotection against myocardial ischemia/reperfusion injury and to explore the underlying mechanisms.

METHODSIsolated perfused rat hearts were subjected to 30 min of ischemia followed by 120 min reperfusion and the cardiac function was evaluated.

RESULTLeft ventricular end-diastolic pressure (LVEDP), left ventricular developed pressure (LVDP) and maximal velocity of contraction and relaxation (+/-dP/dtmax) were improved when U50488H was administered 1 or 24 h before ischemia (P<0.05). Myocardial infarct size, activities of creatine kinase (CK) and lactate dehydrogenase (LDH) in the coronary effluent were lower in the U50488H pretreatment group than those in the control group. Administration of a selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib abolished the late phase of cardioprotection produced by administration of U50488H 24 h before ischemia. Activities of CK and LDH in the coronary effluent were higher in U50488H and celecoxib co-pretreatment group than those in U50488H group. However, administration of celecoxib did not block the early phase of cardioprotection by 1 h treatment of U50488H before ischemia.

CONCLUSIONThe late (but not the early) phase of cardioprotection induced by kappa-opioid receptor agonist might be mediated by COX-2.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer ; pharmacology ; Animals ; Cardiotonic Agents ; pharmacology ; Creatine Kinase ; metabolism ; Cyclooxygenase 2 ; physiology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Male ; Myocardial Infarction ; enzymology ; pathology ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; agonists

OBJECTIVETo analyze the risk factors of heterotopic (HO) ossification after total lumbar disc replacement and probe the preventive strategies for it.

METHODSThe radiographs and clinical data of 78 discs in 65 patients who received artificial lumbar disc replacement (ADR) from April 1998 to December 2003 were analyzed retrospectively by two radiologists and one orthopaedic surgeon and then postoperative HO were graded according to McAfee system. The bony formations in disc spaces, time of HO were found, and range of motion (ROM) of the operated levels were measured on radiographic films. In addition, the risk factors such as preoperative peri-annulus ossification, bony endplates injuries, and subsided or mal-position of the prostheses were also analyzed by Logistic regression analysis.

RESULTSPostoperative HO was found in 10 spaces of 9 cases. Class I of HO were occurred in 7 patients at means 2.1 years postoperatively with normal range of motion preserved. Three of them turned into class II or III with 10 degrees of mean ROM in the following 2.5 years. Another 2 (2/9) cases with preoperative peri-annulus ossification had bridging trabecular bone (class III) between the endplates and 9 degrees of ROM 2 years after surgery, then turned into class IV at 6 years with 0 degrees and 4 degrees of motion in the operated levels. As the risk factors of HO, preoperative annulus ossification (2 cases), bony endplates injuries (5 cases), mal-positioned prostheses (2 cases) and subsided prostheses (2 cases) were found simultaneity with significant positive relation to HO occurred (P < 0.05).

CONCLUSIONSFactors such as preoperative ossification of annulus, endplate injuries, prosthesis subsided and mal-position would have higher risks to have HO occurred after ADR, but ROM of most affected levels are preserved. Strict control indication and avoid all above risk factors can prevent HO occurring effectively.

Adult ; Aged ; Diskectomy ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Ossification, Heterotopic ; etiology ; prevention & control ; Prosthesis Implantation ; adverse effects ; Retrospective Studies ; Risk Factors

OBJECTIVETo investigate the clinicopathological features and prognostic factors of primary clear cell carcinoma of the liver (PCCCL).

METHODSThe clinical data of 41 PCCCL patients who underwent hepatic resection for PCCCL from October 1998 to June 2012 in our department were retrospectively analyzed. There were 31 male and 10 female patients. The median age was 56 years (range, 25 to 80 years), and the diagnosis was confirmed by postoperative pathological examination. The data of 106 well or moderately differentiated non-clear cell hepatocellular carcinoma (HCC) patients and 86 poorly differentiated non-clear cell HCC patients who underwent hepatic resection in the same period in our hospital in the same period were compared. The χ(2) test or Fischer's exact test, as appropriate, was used to compare group frequencies. Survival analysis was estimated by Kaplan-Meier method. Cox proportional hazards model was used in multivariate analysis.

RESULTSThe proportion of fibrous capsule formation in the PCCCL tumors (46%, 19/41) was significantly higher than that of the other two groups (P < 0.05), whereas the PCCCL group had a lower rate of intravascular tumor embolus (2/41) and vascular invasion (1/41) (P < 0.05). The median survival time of PCCCL group was 65 months, the 1-, 3-, 5-year survival rates for PCCCL patients were 90.2%, 67.4% and 42.0%, significantly better than that of poor differentiated NCCHCC group's (82.9%, 33.3%, 7.2%, P < 0.01). However, there were no statistic significant differences between PCCCL group and well or moderately differentiated NCCHCC group (84.7%, 55.7%, 34.4%, P > 0.05). Tumor capsule formation was an independent favorable prognostic factor. In contrast, preoperative serum α-fetoprotein (AFP) level and hepatitis B virus infection were independent unfavorable prognostic factors for PCCCL.

CONCLUSIONSPCCCL is a rare, low degree malignant pathological subtype of HCC. Surgical resection may achieve favorable prognosis and even long-term survival.

Adenocarcinoma, Clear Cell ; blood ; pathology ; surgery ; virology ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; pathology ; surgery ; virology ; Female ; Follow-Up Studies ; Hepatectomy ; Hepatitis B ; Humans ; Liver Neoplasms ; blood ; pathology ; surgery ; virology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate ; alpha-Fetoproteins ; analysis

OBJECTIVETo investigate the prognostic factors of hepatocellular carcinoma.

METHODSThe purpose of this study was to retrospectively analyze the surgical outcomes of hepatocellular carcinoma (HCC) in 832 patients who underwent hepatic resection between February 2002 and June 2010 in the Cancer Hospital of Chinese Academy of Medical Sciences. Post-resection prognostic factors were assessed using a univariate Kaplan-Meier analysis and a multivariate Cox proportional hazards model.

RESULTSThe overall 1-, 3- and 5-year survival rates were 92.0%, 70.2% and 53.6%, respectively. The disease free survival rates (DFS) were 90.2%, 61.5% and 40.5%, respectively. The univariate analysis showed that a better prognosis for overall survival (OS) was associated with asymptomatic presentation, small tumor, single lesion, high-grade histological differentiation, no vascular tumor embolus, negative serum alpha-fetoprotein (AFP), negative serum alkaline phosphatase (ALP), Child-Pugh class A, no ascites, no/mild cirrhosis, new surgical techniques, no blood transfusion, no regional lymph node metastasis, no major vascular invasion, and no extra-hepatic invasion. The multivariate analysis showed that asymptomatic presentation, small tumor, single lesion, no tumor embolus, negative serum alpha-fetoprotein (AFP), no regional lymph node metastasis, no major vascular invasion, no extra-hepatic invasion, no/mild cirrhosis, and surgical techniques are independent factors for a longer overall survival.

CONCLUSIONSThe prognosis of HCC after resection is influenced by a number of factors. Therefore, regularly screening and early diagnosis, applying surgical techniques to minimize the liver injury, and preventing the aggravation of cirrhosis are important measures to improve the overall survival of HCC patients. For those patients with high risk factors of recurrence, routine follow-up is one of the best methods to be recommended.

Alkaline Phosphatase ; blood ; Blood Loss, Surgical ; Bone Neoplasms ; secondary ; Carcinoma, Hepatocellular ; blood ; pathology ; surgery ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; blood ; pathology ; surgery ; Lung Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate ; Tumor Burden ; alpha-Fetoproteins ; metabolism

OBJECTIVETo establish a simplified method of multiplex PCR based on chimeric primers for STR loci.

METHODSA set of chimeric primers and universal primers were designed to carry out a multiplex PCR for amplifying four short tandem repeats(STR) loci. Then the amplified STR loci were detected by polyacrylamide gel electrophoresis and the gels were sliverstained.

RESULTSA quadriplex STR system was developed on the basis of both chimeric and universal primers.

CONCLUSIONThe STR multiplex PCR based on both chimeric and universal primers hss been achieved readily and reproducibly by simple adjustment of the individual primer concentrations. The use of chimeric primers provides a method for primer design that eliminates the multiple optimization steps involved in developing the multiplex PCR.

DNA Primers ; genetics ; Humans ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; methods