ObjectiveTo investigate the osteogenic differentiation potential and the biological features of synovium-derived mesenchymal stem cells (SMSCs) in vitro and to observe the osteogenic capability of the composite scaffolds constructed with SMSCs and hydroxylapatite/chitosan/poly L-latic acid (HA/CS/PLLA) in vivo. MethodSMSCs were separated and cultured with adherent method and enzymatic digestion method. Specific phenotypes of SMSCs were detected by flow cytometry after purification. Then, SMSCs were identified by oil red O staining, alkaline phosphatase (ALP) staining, and alizarin red staining after adipogenic and osteogenic induction, respectively. In vitro experiments:the expressions of osteogenic related genes[osteocalcin (OCN), collagen type I, ALP, and Runx-2] were detected by real-time fluorescent quantitative PCR at 1, 7, 14, 21, and 28 days after osteogenic induction; ALP activities were also determined by ELISA at 1, 3, 5, 7, 9, and 11 days after osteogenic induction; meanwhile, extracellular matrix calcium mineralization was detected by alizarin red S method at 7, 14, 21, and 28 days after osteogenic induction; the normal SMSCs were harvested as control group. In vivo experiments:Twenty-four Sprague Dawley (SD) rats were randomly divided into experimental group (n=12) and control group (n=12) . The 3rd passage SMSCs were seeded on HA/CS/PLLA to construct composite scaffolds, after adhesion for 72 hours in vitro, the composite scaffolds were implanted into the right thigh muscle of 12 SD rats as experimental group; HA/CS/PLLA was implanted into the right thigh muscle of the other 12 SD rats as control group. At 4 and 8 weeks after implantation, the scaffolds were harvested for X-ray film and histological examination to observe ectopic bone formation. ResultsThe positive rates of CD147, CD90, CD105, and CD44 were more than 95%, while the positive rates of CD117, CD34, CD14, and CD45 were less than 10%. Oil red O staining demonstrated red lipid droplets in the cytoplasm, and alizarin red staining showed flaky red calcifications, and cytoplasm was dyed brown by the ALP staining. The mRNA expressions of collagen type I, ALP, and Runx-2 were significantly increased at 7 days after osteogenic induction, and OCN mRNA expression was significantly increased at 14 days after osteogenic induction; ALP activity was significantly higher at 5, 7, 9, 11 days after osteogenic induction in the SMSC-induced group than control group and reached a maximum at 7 days (P<0.05) . Calcium mineralization was significantly enhanced at 14 days after osteogenic induction, and gradually increased with time (P<0.05) ; moreover, it was significantly higher in the SMSC-induced group than control group (P<0.05) . X-ray and histological examination demonstrated that the new bone tissues formed in 2 groups, but bone formation content of the experimental group was significantly more than that of the control group at 4 and 8 weeks after implantation (P<0.05) . ConclusionsSMSCs can be induced into osteoblasts both in vitro and in vivo, so SMSCs might be a promising seed cells for bone tissue engineering.
ObjectiveTo investigate the risk factors of death in patients undergoing continuous renal replacement therapy (CRRT) after cardiac surgery. MethodsWe retrospectively analyzed records of 66 adult patients without history of chronic renal failure suffering acute kidney injury (AKI) following cardiac surgery and undergoing CRRT in our hospital between July 2007 and June 2014. There were 38 males and 28 females with mean age of 59.11±12.62 years. They were divided into a survival group and a non-survival group according to prognosis at discharge. All perioperative data were collected and analyzed by univariate analysis and multivariate logistic regression analysis. ResultsIn sixty-six adult patients, eighteen patients survived with a mortality rate of 72.7%. Through univariate analysis and multivariate logistic regression, risk factors of death in the post-operative AKI patients requiring CRRT included hypotension on postoperative day 1 (B=2.897, OR=18.127, P=0.001), duration of oliguria until hemofiltration (B=0.168, OR=1.183, P=0.024), and blood platelet on postoperative day 1 (B=-0.026, OR=0.974, P=0.001). ConclusionHypotension on postoperative day 1 (POD1) is the predominant risk factor of death in patients requiring CRRT after cardiac surgery, while blood platelet on POD1 is a protective factor. If CRRT is required, the sooner the better.
ObjectiveTo systematically evaluate the efficacy and safety of radiofrequency ablation versus amiodarone in the treatment of atrial fibrillation, so as to provide reference for the chosen of clinical treatment options. MethodsWe searched PubMed, The Cochrane Library (Issue 10, 2014), CNKI, VIP and WanFang data from inception to October 2014 to collect randomized controlled trials (RCTs) comparing radiofrequency ablation versus amiodarone for atrial fibrillation. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then, meta-analysis was performed using RevMan 5.3 software. ResultsA total of 4 RCTs involving 511 atrial fibrillation patients were included. The results of meta-analysis showed that:compared with amiodarone, radiofrequency ablation could reduce the risk of atrial fibrillation recurrence (RR=0.35, 95%CI 0.22 to 0.55, P<0.000 01). There was no significant difference in all-cause mortality (RR=0.97, 95%CI 0.17 to 5.61, P=0.97) between both groups. The incidence of adverse events in the radiofrequency ablation group was 7.7%, and was lower than 12.7% of the amiodarone group, but there was no significant difference between the two groups. ConclusionCurrent evidence shows that, compared with amiodarona, radiofrequency ablation is related to lower recurrence rate and higher efficacy, but there is no difference in the safety between the two interventions. However, due to the limited quality and quantity of included studies, higher quality studies are needed to verify the above conclusion.
ObjectiveTo evaluate the effect of time-related administration of methotrexate (MTX) on neural cell apoptosis in rats after spinal cord injury (SCI) so as to investigate its potential neuroprotective mechanism and appropriate administration time. MethodA total of 120 male Sprague Dawley rats, 247-286 g in weight, were randomly divided into 4 groups (n=30) :sham group (group A), control group (group B), MTX treating group (group C), and MTX prophylaxis group (group D). The SCI model was established in the rats of groups B, C, and D by improved Allen method, and just laminectomy was performed in group A. MTX (0.5 mg/kg) was administered with tail vein injection at 1, 6, 12, 18, and 24 hours after injury in group C, and at 30 minutes before injury and at 6, 12, 18, and 24 hours after injury in group D; the equivalence saline was injected at 1, 6, 12, 18, and 24 hours after injury in groups A and B. Basso-Beattie-Bresnahan (BBB) score was used to evaluate the neural function at 1, 3, 7, 14, and 21 days after injury, HE staining to observe histological changes, immunohistochemical staining and TUNEL method to measure the expression of Caspase-3 and neural cells apoptosis, respectively. ResultsTen rats died during the experiment in groups B, C, and D; 25 rats in each group were included into the experiments at last. BBB score of group A was significantly higher than that of groups B, C, and D at all time points after injury (P<0.05) . BBB score of groups C and D were significantly higher than that of group B at 3, 7, 14, and 21 days (P<0.05) , and BBB score of group D was significantly higher than that of group C at 3, 7, and 14 days (P<0.05) . The histological observation showed normal structure of spinal cord at all time points after injury in group A. While the degree of SCI in group D was lighter than that in groups B and C, and group C was lighter than group B. At 14 days after injury, the degree of SCI in groups B, C, and D tend to keep the same. The number of Caspase-3 and TUNEL positive cells of groups B, C, and D was significantly more than that of group A at all time points after injury (P<0.05) , group B was significantly more than groups C and D (P<0.05) . The number of Caspase-3 positive cells of group C was significantly more than that of group D at 3, 7, and 14 days (P<0.05) . While the number of TUNEL positive cells of group C was significantly more than that of group D at 3 and 7 days (P<0.05) . And the number of Caspase-3 positive cells and TUNEL positive cells was positively correlated in groups B, C, and D (P<0.05) at 1, 3, 7, 14, and 21 days after injury. ConclusionsLow-dose MTX may effectively reduce the degree of the secondary injury of spinal cord by reducing the nerve cell apoptosis. Better effect can be obtained when MTX is used as prevent method than as a way of treatment.
ObjectiveBased on the off-label drug use (OLDU) record application of Alprostadil injection (LipoPGE1) which was the only one rejected in the Guangdong General Hospital in 2013, the interventional measures were carried out to reduce unreasonable off-label drug use of Lipo-PGE1. MethodsMedical orders about OLDU in dosage of Lipo-PGE1 were intervened in through education, communication and monitoring. The situation of drug use was summarized in all departments after intervention through exporting all the medical orders about inpatients' use of LipoPGE1 during hospitalization in August, 2013 to July, 2014 and OLDU incidence in dosage, prescribed daily dose (PDD) and drug use density (DUD) in each department were calculated. The interventional effect was analyzed by comparing with the baseline data. Resultsa) A total of 78 044 medical orders involving 6 426 case-times were analyzed. According to the data of cases, medical orders and drug use amount, the OLDU incidences were 8.68%, 5.87% and 10.53%, respectively, compared with 34.43%, 25.16% and 41.37% before intervention had declined significantly (P < 0.05). OLDU occurred in 69.44% departments (25/36) before intervention and declined to 55.56% (20/36) after intervention. b) OLDU incidences of 22 departments were declined after intervention. There were 2 departments with the OLDU incidence in dose > 20%: ICU (39.68%) and cardiac surgery (32.78%). c) After the intervention, the PDD of the whole hospital fell to 10.52μg from 12.77μg and DUD fell to 8.87 from 15.12. There were 20 departments whose PDDs were off-label and 3 departments whose PDDs were above the average level of the whole hospital after the intervention. The three departments were ICU (13.61μg), cardiac surgery (12.68μg) and rheumatology (11.26μg). ConclusionExtensive publicity and education, targeted communication and regular monitoring and feedback are effective measures to intervene in unreasonable OLDU. After intervention, the phenomenon of off-label drug use of Lipo-PGE1 is improved significantly. This study provides a workable avenue to manage off-label drug use in hospital.
Alpha-glycerophosphate oxidase (α-GPO) from Enterococcus casseliflavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
ObjectiveTo investigate the epidemiology, etiology and prognosis of pneumonia in lung transplantation recipients. MethodsWe retrospectively analyzed the follow-up data of 42 case times (40 patients) of allogenic lung transplantation between March 2005 and August 2014. There were 29 males and 11 females with a mean age of 52.4±13.8 years. There were 32 case times with double lung transplantation, and 10 case times with single lung transplantation. Two patients underwent lung transplantation twice at an interval of 6.5 years and 4.0 years, respectively. ResultsIn 42 case times of lung transplantation, 26 case times had forty-two episodes of pneumonia throughout the follow-up period of median 146 days (range 3 to 2 704 days). Microbiological etiology was established in 36 case times of pneumonia. Bacterial pneumonia (68.1%) was more frequent than fungal (10.6%) and viral pneumonia (8.5%). The cumulative risk of a pneumonia episode increased sharply in the first 30 days after transplantation. A percentage of 38.1% of total pneumonia episodes occurred within 30 days after transplantation, predominately due to Gram negative bacilli. While pneumonia of gram-negative bacilli occurred earliest with a median of 20 days (range 8-297 days). pneumonia caused by viruses (283 days, range 186-482 days) appeared significantly later than gram-negative bacilli, and unknown etiology (44.5 days, range 3-257 days) (P=0.001 and P=0.019, respectively). The survival rate in 1 year, 3 years, and 5 years was 66.1%, 56.3%, and 36.2%, respectively. pneumonia episode within 30 days after lung transplantation was associated remarkably with mortality risk (P=0.03) in lung transplantation recipients. The total blood loss during transplantation procedure and post-transplantation intubation time were associated significantly with early onset of pneumonia (≤30 days) by univariate analysis. ConclusionRecognition of epidemiology, etiology and chronology of post-transplantaion pneumonia has implications relevant for appropriate management and optimal antibiotic prescription in lung transplantation recipients.
ObjectiveBased on the off-label drug use (OLDU) record application of alprostadil injection (Lipo-PGE1) which was the only one rejected in the Guangdong General Hospital in 2013, we retrospectively investigated all the background information of inpatients' OLDU of Lipo-PGE1 in 2012, so as to provide references for intervention of OLDU and effect evaluation. MethodsAccording to OLDU in dose record application of clinical departments, we summarized medical orders about inpatients' use of Lipo-PGE1 during hospitalization in 2012 and analyzed OLDU situation according to drug labels. Then, we summarized situation of drug use in all departments, analyzed OLDU incidence in dose, calculated prescribed daily dose (PDD) and drug use density (DUD) in each department to evaluate the degree of OLDU in dose. Resultsa) A total of 106 576 medical orders involving 8 620 case-times were analyzed. According to the data of cases, medical orders and drug use amount, the OLDU incidences were 34.43%, 25.16% and 41.37%, respectively. Lipo-PGE1 was used in every clinical department in this hospital and OLDU occurred in 69.44% departments (25/36). b) According to the number of medical orders, there were 6 departments with the OLDU incidence in dose > 50% and 5 departments with the OLDU incidence in dose during 50% to 20%. c) The average of PDD of the whole hospital was 12.77 μg. A total of 25 departments' PDDs were off-label, and 10 departments' PDDs were above the average level of the whole hospital. The ICU's PDD was the highest, with 2.35 times more than label dose. d) The PDD in each department was not directly proportional to DUD, which indicated the degree of OLDU in dose was not directly proportional to use intensity. This may be caused by different disease burdens in departments. ConclusionLipo-PGE1 is widely used in the Guangdong General Hospital where OLDU in dose occurs commonly. Since PDD and DUD reflect different contents, the two indicators should be combined to monitor OLDU.
ObjectiveTo explore the value of T-SPOT.TB test in the diagnosis of active tuberculosis and its influencing factors. MethodsFrom July 2010 to November 2012, a total of 289 suspected active tuberculosis patients were enrolled in the study and underwent T-SPOT.TB test. All the patients enrolled were from West China Hospital of Sichuan University. The diagnostic value of T-SPOT.TB applied in determining active tuberculosis was then evaluated. ResultsAccording to the diagnostic criteria, 84 patients diagnosed with active tuberculosis were eligible for analysis and enrolled as a tuberculosis group, and 156 patients were enrolled as a control group. The sensitivity of T-SPOT.TB test was 83.3%, while the specificity was 80.1%. Both univariate and multivariate analyses showed the characteristics of patients such as general conditions (eg. age, sex) and basic diseases (eg. immunosuppression condition, malignant tumour) were not the risk factors of false-positive or false-negative result of T-SPOT.TB. ConclusionT-SPOT.TB test for the diagnosis of active tuberculosis has high sensitivity and specificity, with important value referred for diagnosing suspected active tuberculosis patients.
ObjectiveTo compare the cost-effectiveness of warfarin and enoxaparin overlapping treatment for the prevention of venous thromboembolism (VTE) or pulmonary embolism (PE) in patients with nephrotic syndrome (NS). MethodsA decision tree model was constructed. The efficacy data applied in our decision tree were from clinical data, and the cost data was based on the hospitalization cost of 103 patients with nephrotic syndrome in Guangdong Provincial People's Hospital from 2013 to 2014, State Development and Reform Commission pricing and literature report. The one-way sensitivity analyses was conducted to analyze the stability of test. ResultsIn base case, the cost and cost-effective ratio of warfarin and enoxaparin overlapped treatment for 3 days were 10305.49 yuan and 31607.15, respectively. While those overlapped treatment for 4 days were 8849.36 yuan and 20896.46, overlapped treatment for 5 days and above were 9494.29 yuan and 21659.95, respectively. The incremental cost-effectiveness ratio of 4 days versus 5 days and above was 5600.96. The cost-effective ratio of 4 days was lower but the incremental cost-effectiveness ratio of it was higher. The sensitivity analysis showed the test result was stable. ConclusionCost-effectiveness analysis shows that warfarin and enoxaparin overlapping treatment for 4 days in patients with nephrotic syndrome has cost-effective advantage. Due to the limited sample size of our study, the above conclusion should be proved by more large-scale high-quality clinical studies.