ObjectiveTo study the clinical effect of improved decortication in treatment of tuberculous empyema as well as the safety of operation.MethodsEighty-two cases who diagnosed as tuberculous empyema by pathology and operation from January 2007 to September 2010 were selected including improved decortication 44 cases,total empyema decortication 28 cases,thoracoplasty 10 cases.The operation time,peri-operative bleeding,postoperative complication and lung function recovery after 6 months operation were followed-up.ResultsAll the patients were in good recovery and there was no death in the operation for 1 year.The peri-operative bleeding in improved decortication was less than that in total empyema decortication and thoracoplasty [(56.23 ± 15.56) ml vs. (78.65 ± 23.14) and (66.92 ± 19.83) ml],and there was significant difference among them(P＜ 0.01 ).There was no postoperative complication in improved decortication,but 2 cases (7.1％,2/28) of partial lung recruitment maneuvers in total empyema decortication,1 case ( 10.0％,1/10) of extensive staxis with selective surgery in thoracoplasty.The proportion of forced vital capacity (FVC),forced expiratory volume in one second (FEV1),peak expiratory flow (PEF) turning to normal after 6 months operation in improved decortication [95.5％(42/44),93.2％(41/44),97.7％(43/44)]were higher than those in total empyema decortication[ 75.0％ (21/28 ),78.6％ (22/28),85.7％ (24/28) ]and thoracoplasty [ 80.0％ (8/10),90.0％ (9/10),80.0％ ( 8/10) ],and there was significant difference among them (P ＜0.01).There was no significant difference in the operation time among them(P ＞0.05).ConclusionImproved decortication has remarkable superiority in curing tuberculous empyema especially it is small in traumatic with less blood loss,fewer postoperative complications,lung function recovery rapidly after operation.

Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by silicone tube idled with valproic acid (VPA). Methods In present research we demon-strate the effect of VPA on peripheral nerve regeneration and recovery of motor function following sciatic nerve transaction in rats. An 8-mm sciatic nerve deficit was created in a rat mode land bridged by a 1-cm silicone tube.Then, 10 lad of 8% VPA were perfused into the silicone chamber in the VPA group. The same volume of normal saline was delivered in the control group. Results Each animal was observed sciatic nerve function index (SFI) at 2-week intervals and studied electrophysiology at 4-week intervals for 12 weeks. Histological and morphometrical analyses were performed at the end of the experiment, 12 weeks after operation. Using the digital image-analysis system, thickness of the myelin sheath was measured, and total numbers of regenerated axons were counted. There was a significant difference in SFI, electrophysiological index (motor-nerve conduct velocity, MCV), and morphometrical results (regenerated axon number and thickness of myelin sheath) in nerve regeneration between the VPA group and controls (P ＜ 0.05). Conclusion The results demonstrated that VPA is able to enhance sciatic nerve regeneration in rats, suggesting the potential clinical application of VPA for the treatment of peripheral nerve injury in humans.

Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by a new PRGD/PDLLA/VPA composite conduit.Methods In this study from February,2012 to March,2014,PRGD/PDLLA/VPA nerve conduits were tested in the rat sciatic nerve transection model.At different periods after operation,its ability to promote nerve regeneration was evaluated by sciatic functional index(SFI),electrophysiology (CMAPs,NCVs) and histologic assessment.Forty rats were randomly divided into 4 groups (n =10),group A:PRGD/PDLLA/VPA,group B:PDLLA/VPA,group C:PRGD/VPA and group D:autograft.Results At 12 weeks after surgery,the SFI value of group A (-45 ± 3.19)and group D (-42 ± 3.01)were significantly higher than those of group B(-79 ± 3.06) and group C(-72 ± 2.07)(P ＜ 0.05);The CMAPs of group A (24.89 ± 5.01) and group D (25.39 ± 5.63) were significantly higher than those of group B(14.88 ± 3.11) and C(15.00 ± 5.54);the NCVs of group A (31.42 ± 2.43) and group D (31.50 ± 2.16) were significantly higher than those of group B (20.11 ± 2.39) and group C(21.00 ± 2.13)(P ＜ 0.05).At 12 weeks after surgery,the numbers of regenerated nerve in the tube of group A (258 ± 6.18) and D(259 ± 5.59) were significantly higher than those of group B (231 ± 5.00) and group C(230 ± 5.07)(P ＜ 0.05).There was no significant difference between groups A and D(P ＞ 0.05).Conclusion These results illustrated that this new PRGD/PDLLA/VPA conduit could significantly facilitate the regeneration of short nerve defect and recovery of motor nerve,which provides a new thought for treatment of peripheral nerve injury.

BACKGROUND:Studies have found that combination of two of chitosan (CS), nano-hydroxyapatite (nHA) and poly(lactide-co-glycolide) (PLGA) can improve the mechanical properties and biocompatibility of the composite stent in certain extent as wel as improve osteogenic differentiation of the cels, but there is a certain distance from the ideal bone tissue engineering scaffolds.
OBJECTIVE:To study biocompatibility and osteoinductive activity of nHA/CS/PLGA scaffolds with different proportions in vitro.
METHODS: nHA/CS/PLGA scaffolds were prepared at mass ratio of 10:10:80, 10:20:70, 20:10:70 respectively by particle leaching method. And human bone marrow stem cels (hBMSCs) were co-cultured with these scaffolds in vitro. Adhesion, proliferation, and osteoinductive activity of these scaffolds were examined qualitatively and quantitatively by growth curve of hBMSCs on scaffolds. Gene expression of alkaline phosphatase activity and osteocalcin was detected by RT-PCR.
RESULTS AND CONCLUSION: hBMSCs could be attached, proliferated, and osteoinduced better on the nHA/CS/PLGA scaffold with the mass ratio of 20:10:70, compared to the other two groups of scaffolds. The differences were significant statisticaly (P< 0.05). Alkaline phosphatase and osteocalcin expressions were respectively higher in the scaffold with the mass ratio of 20:10:70 after 9-27 days of co-culture and 15-27 days of co-culture, in comparison with the other two groups of scaffolds. These findings indicate that the nHA/CS/PLGA scaffolds with the mass ratio of 20:10:70 demonstrated preferable biocompatibility and osteogenic inductivity, which is expected to be a promising scaffold material for bone tissue engineering.

Objective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.

To establish a LC-MS/MS method to determine the concentrations of liquiritin, glycyrrhizin, glycyrrhetinic acid, amygdalin, amygdalin prunasin, ephedrine, pseudoephedrine and methylephedrine of Maxing Shigan decoction in rat plasma, and study the differences on their pharmacokinetic process in normal rats and RSV pneumonia model rats. After normal rats and RSV pneumonia model rats were orally administered with Maxing Shigan decoction, the blood was collected from retinal vein plexus of different time points. Specifically, tetrahydropalmatine was taken as internal standard for determining ephedrine, while chloramphenicol was taken as internal standard for determining other components. After plasma samples were pre-treated as the above, the supernatant was dried with nitrogen blowing concentrator and then redissolved with methylalcohol. The chromatography was eluted with mobile phase consisted of acetonitrile and 0.1% formic acid solution in a gradient manner. ESI sources were adopted to scan ingredients in ephedra in a positive ion scanning mode and other ingredientsin a negative ion scanning mode. The multiple-reaction monitoring (MRM) method was developed the plasma concentration of each active component. The pharmacokinetic parameters of each group were calculated by using Win-Nonlin 4.1 software and put into the statistical analysis. The result showed the plasma concentration of the eight active ingredients, i.e., liquiritin, glycyrrhizin, glycyrrhetinic acid, amygdalin, amygdalin prunasin, ephedrine, pseudoephedrine and methylephedrine within the ranges of 1.04-1040, 1.04-1040, 0.89-445, 1.05-4200, 1.25-2490, 0.3-480, 0.3-480, 0.3-480 microg x L(-1), with a good linearity and satisfactory precision, recovery and stability in the above ingredients. After modeling, except for glycyrrhetinic acid whose pharmacokinetic parameters were lacked due to the data missing, all of the rest components showed significant higher Cmax, AUC(0-1) and lower clearance rate (CL) than that of the normal group, indicating the increase in absorption in rats in the pathological state by reducing the clearance rate. The method is accurate and sensitive and so can be used to determine the plasma concentrations of the eight active ingredients in Maxing Shigan decoction. RSV pneumonia-infected rats absorbed more ingredients in Maxing Shigan decoction.
Animals ; Chromatography, High Pressure Liquid ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacokinetics ; Male ; Pneumonia, Viral ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Respiratory Syncytial Virus Infections ; drug therapy ; metabolism ; Tandem Mass Spectrometry

Objective:Imatinib is extensively used as a first-line therapeutic agent for patients with chronic myeloid leukemia (CML) at the chronic phase (CP). Although CML patients undergoing imatinib treatment are enrolled mainly in the Glivec International Patient Assistance Program (GIPAP) in China since 2003, limited data have been reported on the long-term outcome of these patients. This study aims to compare the treatment response and prognosis of CML-CP patients who received different treatments from January 2003 to December 2013 in the First Affiliated Hospital of Nanchang University. Methods:A total of 295 patients were enrolled, includ-ing 185, 30, 50, and 30 patients for imatinib, interferon-alpha (IFN-α) plus Ara-C, hydroxycarbamide (HU), or allogeneic hematopoietic stem cell transplantation (Allo-HSCT) treatments, respectively. Results:Patients in imatinib and Allo-HSCT groups achieved excellent complete hematologic remission (CHR) (i.e., 96.7%vs. 96.7%), complete cytogenetic response (CCyR) (i.e., 89.7%vs. 93.3%), and com-plete molecular remission (CMoR) (i.e., 49.7%vs. 83.3%, P=0.001). However, significantly low rates of CHR, CCyR, McyR, and CMoR were observed in IFN-αand HU groups. Moreover, patients from imatinib group showed longer overall survival (OS) time than patients from other groups (P<0.001), even patients in Allo-HSCT group (10-year OS, 89.0%vs. 67.0%, P<0.001) because of high risk of Allo-HSCT-related complication. Multivariate analysis showed that receiving imatinib treatment (HR=5.267, 95%CI:1.054-1.940, P=0.022) and achieving CCyR (HR=9.541, 95%CI:1.692-10.513, P=0.002) were independent predictors for OS. Conclusion:Imatinib treatment may be an optimal first-line choice for Chinese patients with CML-CP who have not received any previous treatments.

OBJECTIVETo further study the mechanism of the inhibitory effect of interferon beta-1a (IFN beta-1a) on the activation of human hepatic stellate cell (HSC) LX-2, and to analyze the differences on the protein expression in LX-2 induced by I IFN beta-1a.

METHODSCultured LX-2 cells were treated with 2000 U/ml IFN beta-1a for 48 h. Two-dimensional gel electrophoresis (2-DE) was performed to compare protein patterns of the control (untreated) and IFN beta-1a treated LX-2 and for quantitative and qualitative analyses of protein expression. A rat liver fibrosis model was established and the rats were sacrificed and their various tissues were obtained for the same analyses. Western blotting and RT-PCR were used to validate the expression of the changed proteins after treatment of IFN beta-1a in LX-2 cells and of various tissues of the rats.

RESULTS708 +/- 25 spots were detected in control LX-2 cells and 804 +/- 32 spots in IFN beta-1a-treated LX-2 cells. A match rate of 73%-82% was achieved. The results also showed that 31 protein spots displayed quantitative changes in expression after IFN beta-1a treatment. Of the 31 spots, 21 proteins were identified, of which, one was newly found, two were enhanced in abundance and 18 showed lower expressions. The newly found protein was glia maturation factor beta (GMF beta). The treatment of LX-2 with IFN beta-1a increased the production of GMF beta(GMF beta) protein in comparison with the untreated cells (t=1.81, P < 0.01). The expression of GMF beta protein (1.81 vs 0.10) and mRNA (0.85 vs 0.12) were more in the normal liver tissues than in the cirrhotic liver tissues (t=2.53, 2.13 respectively, P < 0.01). The expressions of GMF beta protein and mRNA were weak in rat heart and lung tissues, however, they were strong in rat liver, kidney, spleen and brain tissues (t=1.91, 1.94 respectively, P < 0.01).

CONCLUSIONThere is a significant difference of protein expression levels between IFN beta-1a untreated and treated LX-2 cells. These proteins, especially GMF beta, may be involved in an inhibition process of IFN beta-1a on activation and apoptosis of LX-2 cells. This proteome study may be useful in further studies of the relationship of IFN beta-1a treatment and human liver diseases.

Animals ; Cell Line ; Female ; Glia Maturation Factor ; metabolism ; Hepatic Stellate Cells ; metabolism ; Humans ; Interferon beta-1a ; Interferon-beta ; pharmacology ; Liver ; cytology ; Liver Cirrhosis ; metabolism ; Proteome ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line.

METHODSAstrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 min; carbenoxolone (CBX)+IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca(2+)+HS group, astrocytes were pre-incubated with Ca Ca(2+) (1,000 micromol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM).

RESULTS(1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 min (P < 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P< 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca(2+)+HS group were significant lower than that in the HS group (P < 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells.

CONCLUSIONHS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.

Analysis of Variance ; Animals ; Animals, Newborn ; Astrocytes ; drug effects ; metabolism ; Calcium ; pharmacology ; Carbenoxolone ; pharmacology ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Flow Cytometry ; Gene Expression Regulation ; drug effects ; Glial Fibrillary Acidic Protein ; metabolism ; Glutamic Acid ; metabolism ; Hypothalamus ; cytology ; Rats ; Saline Solution, Hypertonic ; pharmacology ; Time Factors

OBJECTIVESTo study the effects of rat endothelial progenitor cell (EPC) transplantation on hepatic fibrosis in carbon tetrachloride (CCl4) induced hepatic fibrosis rats.

METHODSHepatic fibrosis was developed in 24 healthy female SD rats by feeding them 25% CCl4/olive oil for 8 weeks. Eight of them were sacrificed at the end of the 8 weeks. The rats were subdivided into a EPCs transplanting group (n=8) and a saline control group (n=8). After the EPCs were isolated and cultured for 9 days, the cells were injected into the portal veins of the rats in the EPCs transplanting group. Four weeks later all of the rats were sacrificed. The blood biochemical parameters from the serum were examined. The degree of liver fibrosis was evaluated by reading Masson staining liver slides and by detecting the expression of a-SMA and collagen III.

RESULTSCompared with the saline control group, hepatic activity index (HAI), levels of ALT, AST and TBil in the serum were all lower in the EPCs transplanting group, but the level of Alb was higher and the expression of a-SMA and collagen III were lower. Compared with the 8 week hepatic fibrosis group, the levels of ALT, AST and TBil in the serum of the EPCs transplanting group were all lower. In the saline control group, the serum levels of ALT, AST and TBil were higher, the level of Alb was lower, and the expressions of a-SMA and Collagen III were higher.

CONCLUSIONIn hepatic fibrosis rats, transplantation of rat EPCs could minimize the hepatic fibrosis process and the injuries.

Animals ; Carbon Tetrachloride ; Endothelial Cells ; cytology ; Female ; Liver Cirrhosis, Experimental ; chemically induced ; pathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation