Objective:To establish a determination method for the content and entrapment efficiency of ropivacaine hydrochloride-loaded multivesicular liposomes. Methods: The separation of the multivesicular liposomes from the free drug was achieved by low-speed centrifugation. The concentration of ropivacaine hydrochloride in the supernatant and the multivesicular liposomes was determined by HPLC, and the entrapment efficiency was calculated. Results: The linear range of ropivacaine hydrochloride was 1. 0-80. 0μg· ml-1(r=0. 999 8). The average recovery was 99. 95% and RSD was 0. 72%(n=9). The content and entrapment efficiency of three batches of ropivacaine hydrochloride-loaded multivesicular liposomes was within the range of 99. 1%-100. 3% and 80. 06%-82. 14%, respectively. Conclusion:The method is simple and accurate, and can be used in the determination of content and entrapment efficien-cy of ropivacaine hydrochloride-loaded multivesicular liposomes.

Aim To investigate the protective effect of different concentrations of isoflurane preconditioning in isolated rat hearts and the relationship between this effect with changes of endogenous antioxidant enzymes.Methods The isolated SD rat hearts perfused on Langendorff apparatus were randomly divided into 6 groups(n=14):control group(CON);1.44 MAC isoflurane control group(ISO);ischemia-reperfusion group(I/R);3 isoflurane preconditioning groups(I1,I2,I3).All hearts were made globally ischemic for 30 min followed by 60 min reperfusion except CON and ISO.The hearts of ISO and I1,I2,I3 were perfused with K-H solution saturated with 0.72 MAC or 1.08 MAC or 1.44 MAC isoflurane respectively for 20 min followed by 5 min washout after equilibrium.The cardiac function variables were measured after equilibrium,immediately before ischemia,at the end of 30 min and 60 min reperfusion.The infarct size and activities of the endogenous antioxidant enzymes were measured at the end of reperfusion.And the activities of enzymes in I2 group were measured after equilibrium,immediately before ischemia,immediately after ischemia and at the end of 30 min reperfusion.Results Ischemia/reperfusion significantly increased LVEDP and decreased LVDP,dp/dtmax,dp/dtmin and HR(P

Objective To investigate the effect of intrathecal γ-aminobutyric acid transporter-1 ( GAT-1 )small interfering RNA (siRNA) on neuropathic pain in rats. Methods Male SD rats weighing 200-250 g were studied. The experiment was performed in 3 parts. Part Ⅰ Twenty rats were randomly divided into 5 groups ( n =4 each): GAT-1 siRNA-1 group, GAT-1 siRNA-2 group, GAT-1 siRNA-3 group, negative control siRNA group and DEPC treatment group. Two days after ligation of sciatic nerve, intrathecal siRNA 2 μg or equal volume of DF-PC was injected once a day for 3 consecutive days. The rats were killed and the lumbar segment of the spinal cord was removed at 2nd day after the last intrathecal injection for determination of the expression of GAT-1 in the spinal dorsal horn by Western Blot. Part Ⅱ Thirty rats were randomly divided into 3 groups ( n = 10 each): GAT-1 siRNA-3 + lipo2000 group, GAT-1 siRNA-3 mismatch siRNA + lipo2000 group, and DEPC treatment + lipo2000group. Paw-withdrawl threshold (PWT) to thermal and mechanical stimulation was measured before ligation of sciatic nerve, 3 days after ligation of sciatic nerve and at 1, 3, 5, 7 and 10 days after consecutive administration for 3 days. Part Ⅲ Eighty-four rats were randomly divided into 3 groups as described in Part Ⅱ ( n = 28 each). Four rats were killed at each time point and the lumbar segment of the spinal cord was removed for determination of the expression of GAT-1 in the spinal dorsal horn by Western blot. Results PWT to thermal and mechanical stimulation was significantly inreased and the GAT-1 expression was down-regulated after the injection of GAT-1 siRNA.Conclusion Intrathecal GAT-1 siRNA can reduce the neuropathic pain by inhibiton of up-regulation of the GAT-1 expression in the spinal dorsal horn in rats.

Objective To investigate the role of gamma-aminobutyric acid transporter-1 (GAT-1) in the spinal cord in a rat model of bone cancer pain.Methods Eighty female SD rats weighing 150-180 g were randomly divided into 5 groups (n =16 each): sham operation group(group Ⅰ ),bone cancer pain group(group Ⅲ ),sham operation+ NO-711 group(group Ⅲ ),Ⅳ group BCP + NO-711 group(group Ⅳ ) and BCP + vehicle group (group Ⅴ ).Bone cancer pain was induced by inoculating Walker-256 mammary gland carcinoma cells into medullary cavity of tibia.NO-711 (20 μg,10 μl) was administered intrathecally once a day for 3 consecutive days from the 14th day after operation.Mechanical withdrawl threshold (MWT) of mechanical stimulus was determined the day before operation and at days 3,5,7,10,14 and 16 after operation.The animals were sacrificed on the 16th day after operation,and then the spinal cords were removed for determination of the expression of GAT-1 and double immunostaining of GAT-1 and glial fibrillary acidic protein (GFAP,astrocyte marker).Results MWT were significantly decreased in groups Ⅱ,Ⅳ and Ⅴ as compared with groups.Ⅰ and Ⅲ.The expression of GAT-1 significantly up-regulated in groups Ⅱ,Ⅴ as compared with groups Ⅰ and Ⅲ.NO-711 significantly increased MWT,while decreased the expression of GAT-1 in group Ⅳ compared with groups Ⅱ and Ⅴ.The expression of GAT-1 up-regulation appeared colocalizes with in astrocytes activation in spinal dorsal horn.Conclusion The up-regulation of expression of GAT-1 in spinal cord is involued in the development and maintenance of bone cancer pain,which may be related to the astrocytes activation.

Objective To explore X-ray measurement of the subacromial space and to determine the normal range of it.Methods Shoulder joints in 188 healthy adults were examined with routine X-ray on the anterior-posterior position and the anterior lateral part of acromion to the intercondylar line of humerus(h1),as well as the most lateral part of acromion to intertubercular sulcus of humeral head(h2) were measured and observed.Results The 95% reference ranges in h1,h2 were 6.4～10.4 mm and 11.0～17.6 mm,respectively.Conclusion Both h1 and h2 can correctly reflect the bony distance of subacromial space,which is of significant meaning for guiding clinical diagnosis and therapy.

Aim To explore the effect of propofol preconditioning on cardiocyte apoptosis and cytochrome C release from mitochondria during hypoxia/reoxygenation in isolated rat hearts.Methods Fifty male Sprague-Dawley rats weighing 250～300 g were randomly divided into 5 groups(n=10 each):control group(C);Dimethyl sulfoxide(DMSO)preconditioning group(D);3 propofol preconditioning groups with 25 ?mol?L-1(P1)、50 ?mol?L-1(P2)、100 ?mol?L-1(P3)propofol respectively.The isolated rat hearts were retrogradely perfused via aorta with K-H solution on Langendorff apparatus.The isolated hearts were made hypoxia for 30 minutes followed by 60 minutes reoxygenation in each group.The D,P1,P2,P3 groups were preconditioned by perfusing 10 min K-H solution containing 20 ?mol?L-1 DMSO and 25,50,100 ?mol?L-1 propofol respectively and then followed by 5 min K-H solution reperfusion before hypoxia.The preconditioning procedure was repeated twice.The cardiac functional variables were recorded after equilibration(baseline values),immediately before hypoxia,at the end of 30 min and 60 min reoxygenation.Apoptotic myocardial cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling(TUNEL)and the level of cytochrome C expression in myocardial cytosol and mitochondria was measured by Western blot at the end of reoxygenation.Results At the end of 30 min and 60 min reoxygenation,LVEDP was significantly lower and LVDP was significantly higher in P1,P2,P3 groups than in D group(P

0.05).Inside experimental area,pain scores were decreased in both intradermal 0.25% lidocaine and saline injection group(P

Objective To evaluate interleukin-27 ( IL-27 ) as a sepsis diagnostic biomarker in critically ill adults with sepsis. Methods A retrospetive study was conducted. A total of 176 systemic inflammatory response syndrome ( SIRS ) patients in Department of Critical Care Medicine of Xinxiang Medical College First Affiliated Hospital from March to November in 2014 were enrolled. The patients were divided into no sepsis group ( n=66 ), pulmonary originated sepsis group ( n=65 ), and non-pulmonary originated sepsis group ( n=45 ). Plasma IL-27 and procalcitonin ( PCT ) were determined with enzyme linked immunosorbent assay ( ELISA ). Receiver operating characteristic curve ( ROC ) and classification and regression tree methodology was used to evaluate diagnostic biomarker performance. Results The proportion of patients in pulmonary original sepsis group whose body temperature in line with SIRS criteria was significantly higher than no sepsis group ( 66.2%vs. 44.5%, P<0.05 ), and they were easy to suffer from tumor ( 44.6%vs. 22.7%, P<0.05 ). The proportion of patients in non-pulmonary originated sepsis group whose white blood cell count in line with SIRS criteria was significantly higher than no sepsis group ( 68.9%vs. 42.7%, P<0.05 ). It indicated that patients in pulmonary originated sepsis group and non-pulmonary originated sepsis group were more in line with SIRS criteria compared with no sepsis group. It was shown by ROC curve that IL-27 and PCT was not effective in discriminating sepsis among unselected patients showing symptoms and signs of SIRS. The area under the curve ( AUC ) was 0.59 [ 95%confidence interval ( 95%CI )=0.49-0.65 ] and 0.61 ( 95%CI=0.55-0.71 ). According to the further analysis from different infection sources, the highest AUC was 0.71 ( 95%CI=0.59-0.79 ) for IL-27 in patients with a non-pulmonary originated sepsis. A decision tree incorporating IL-27, PCT, and age had an AUC of 0.78 ( 95%CI = 0.71-0.87 ) in patients with a non-pulmonary originated sepsis, which was higher than IL-27 [ 0.71 ( 95%CI = 0.59-0.79 ) ] or PCT [ 0.65 ( 95%CI = 0.57-0.78 ) ]. Compared to that of pediatric cohort with sepsis, lower expression of IL-27 was found in adult patients. Conclusions IL-27 performed overall poorly as a sepsis diagnostic biomarker in adults. IL-27 may be a more reliable diagnostic biomarker for sepsis in children than in adults. The combination of IL-27 and PCT can reasonably estimate the risk of sepsis in subjects with a non-pulmonary originated sepsis.

Objective To analyze the recurrence rate of intubation and increase of ventilator support rate within 24 hours after using fiberoptic bronchoscopy (FOB) in critically ill patients with hypoxemia complicated with respiratory failure, and to approach the feasibility of FOB in such patients.Methods A prospective study was conducted, including 200 critically ill patients with acute respiratory failure using FOB [oxygenation index (PaO2/FiO2) ≤ 300 mmHg (1 mmHg = 0.133 kPa)] admitted to the intensive care unit (ICU) of the First Affiliated Hospital of Xinxiang Medical College. The rates of intubation and increased ventilatory support and the reasons for bronchoscopy related complications after using FOB 24 hours were recorded, the main risk factors leading to these changes and complications were analyzed and screened by logistic regression analytic method.Results Within 24 hours after using FOB for 200 patients with respiratory failure, an increase in mechanical ventilatory support was required in 68 patients (34%) of that 28 (14%) led to endotracheal intubation. With the extension of time, the rates of intubation and ventilatory support showed a tendency of elevation, the rise in ventilatory support rate being faster. The reasons for bronchoscopy related complications after FOB consisted of cardiovascular disease (41%), coronary artery disease (17%), chronic obstructive pulmonary disease (COPD, 17%), chronic restrictive pulmonary disease (10%), immunity suppression (54%), malignant neoplastic hematologic disorder (20%), acquired immune deficiency syndrome (AIDS, 12%), solid organ transplantation (3%), solid tumor (10%), corticosteroid therapy (25%), immunosuppressive drug (16%), diabetes (15%), chronic renal failure (14%), swallowing nerve injury (37%), anticoagulant therapy (19%), antiplatelet therapy (13%). In the patients with occurrence of COPD or immunosuppression, the rate of invasive ventilation used was significantly higher than that without using invasive ventilation [COPD: 35% (10/28) vs. 14% (24/172),χ2 = 8.081,P = 0.004; immunosuppression: 75% (21/28) vs. 50% (86/172),χ2 = 6.051,P = 0.014]. The logistic regression analysis showed that the occurrence of COPD or immunosuppression was obviously related to whether the intubation being necessary or not [COPD: odds ratio (OR) = 5.200, 95% confidence interval (95%CI) = 1.500 - 17.700,P = 0.006; immunosuppression:OR = 5.300, 95%CI =1.600 - 17.100,P = 0.004].Conclusions In patients with hypoxemia using FOB, they often require addition of mechanical ventilatory support, but the intubation rate is not high. Under the ventilatory support, FOB has certain feasibility for treatment of critically ill patients with hypoxemia and acute respiratory failure.

Objective To investigate the clinical significance of neutrophil gelatinase-associated lipocalin (NGAL),monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-like apoptosis weak inducer (TWEAK) level in body fluids of lupus nephritis (LN) patients..Methods Levels of NGAL,MCP-1 and TWEAK in the serum and urine of 51 patients with SLE,including 27 LN cases and 24 non-LN cases,were detected by enzyme-linked immunosorbent assay; and the clinical data were collected.Student's t test,Mann-Whitney U test,Pearson's correlations and Spearman's rank correlations were used for statistical analysis.Results (①) The level of serum NGAL in the LN group (122±70) ng/ml was higher than that in the non-LN group (56 ± 34) ng/ml (P＜0.01).There was a positive correlation (r=0.408,0.431,0.339,0.403 and 0.585,P＜0.05) between the serum NGAL and general SLEDAI score,extra-renal SLEDAI score,renal SLEDAI score,the levels of 24-hour urine protein and serum creatinine.There was a negative correlation (r=-0.396,P＜0.01) between the serum NGAL and serum C3 level.There was a positive correlation (r=0.719,P＜0.01) between urinary NGAL levels and chronic index (CI).② The level of serum MCP-1 in the LN group (284 ± 113) pg/ml was higher than that in the non-LN group (173± 69) pg/ml,(P＜0.01).The level of urinary MCP-1 in the LN group (M=1154,P75 41 178,P25 341) pg/mg creatinine was higher than that in the non-LN group (M=456,P75 714,P~ 114) pg/mg creatinine (P＜0.01).There was a positive correlation (r=0.340,0.416,0.385,0.574 and 0.654,P＜0.05) between the serum MCP-1 level and the overall SLEDAI score,extra-renal SLEDAI score,renal SLEDAI score,24-hour urinary protein excretion level and serum creatinine levels.There was a negative correlation (r=-0.458,P＜0.01) between the serum MCP-1 level and serum C3 levels.There was a positive correlation (rs=0.448,0.429,0.459,0.412,0.375 and 0.419,P＜0.05) between urinary MCP-1 level and total SLEDAI score,extra-renal SLEDAI score,renal SLEDAI score,serum ds-DNA antibodies,24-hour urinary protein excretion level and serum creatinine levels.There was a significant positive correlation (r=0.689,P＜0.01) between urinary MCP-1 levels and CI.(③) TWEAK was detected in the urine of 6 cases of LN.Conclusion The urinary NGAL and MCP-1 levels are elevated in patients with chronic damage,which suggestes chronic renal injury.It can be used to monitor the LN disease activity.The blood levels of NGAL and urine MCP-1 are significantly higher in LN.The clinical significance of TWEAK needs further study.