A new wood-degrading fungus Monodictys asperospera(Cooke & Massee) Ellis with a high level of laccase production was chosen to study.This laccase was purified by ammonium sulfate precipitation,DEAE-cellulose and sephacryl S-300.Purification of about 8.1 fold was achieved with an overall yield of 5.7%.Its molecular weight was estimated to be about 77 kD.The optimum temperature and pH of the lac-case activity were 55?C and 6.0,respectively.Kinetic studies of the laccase showed that the Km and the Vmax for using syringaldazine as substrate was 0.163 mmol/L and 0.194 mmol/(L.min),respectively.The carbo-hydrate content was 18.14%.In addition,it was found that laccase activity was significantly inhibited by Cu2+.

12 strains of H2-producing bacteria were isolated and purified from anaerobic sludge, aerobic sludge and river bottom sludge by Hungate method. A new species of high efficient hydrogen production bacterium Enterococcus sp. LG1 (registration number: EU258743 ) was studied deeply. It was showed that the Enterococcus sp. LG1 was an anaerobic and Gram-negative bacterium. Sequence analysis of this type of clones showed that it was affiliated with the genus Enterococcus and it was not reported yet in other paper at present. Meanwhile, batch tests of anaerobic fermentative hydrogen production by Enterococcus sp. LG1 were investigated by using sterilization pretreated sludge as substrate. The changes of soluble COD, protein, carbohydrate and pH value during hydrogen fermentation were monitored. It was found that only hydrogen and carbon dioxide were produced by this strain and no methane was detected during fermentation. The maximal hydrogen yield was 36.48 mL/g TCOD and the hydrogen concentration in the gas phase was 73.5%. The Enterococcus sp. LG1 was a butyrate fermentation bacteria analyzed by metabolites.

In this study, the human umbilical vein endothelial cell model was used to study the regulating effect of lipophilic components in Salvia miltiorrhiza on angiogenesis, and explore its possible mechanism. The cell model was established to determine the effect of lipophilic components in S. miltiorrhiza on the proliferative activity and migration capacity of endothelial cells. Then the realtime fluorescence quantification PCR technology was applied to detect the changes in the gene expressions of angiogenesis-related cytokines VEGF-A, VEGF-C and MMP-9. The results showed that 5 mg x L(-1) lipophilic components in S. miltiorrhiza could inhibit the proliferation and migration of endothelial cells, and reduce the expression of VEGF-A and MMP-9 genes. It indicated that lipophilic components in S. miltiorrhiza may inhibit the proliferation and migration of endothelial cells by inhibiting the expression of VEGF-A and MMP-9 genes, so as to show the inhibitory effect on angiogenesis.
Angiogenesis Inhibitors ; chemistry ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo explore effectiveness and safety of segmental anterior cervical decompression in treating multi-level cervical myelopathy.

METHODSTwenty-four patients with four levels of cervical myelopathy were treated with segmental anterior cervical decompression (reservation of middle vertebrae, bone graft and plate-screws fixation). Among patients, there were 15 males and 9 females aged from 47 to 75 (averaged 57.9) years old. Preoperative, postoperative at 1 week and the latest following-up AP and lateral X-rays were used to observe bone union, displacement of implant, adjacent segment degeneration, changes of Cobb angle of fusion segment. JOA scoring were applied for evaluate recovery of nerve function.

RESULTSAll operations were completed successfully, 2 cases ocurred hoarseness, and improved after treated symptomatically. Nineteen patients were followed up from 3.1 to 5.3 years with an average of 3.9 years. Bone union time ranged from 3 to 7 (averaged 4.5) months. No screw loosening and displacement occurred. Nine patients occurred titanium mesh subsidence in different degrees, and 4 of them subside >3 mm; four patients ocurred adjacent segment degeneration. Postoperative Cobb angle of fusion segment at 1 week (10.40±2.94)° was improved from preoperative (5.76±4.16)°, but decreased at the latest follow-up (8.57±2.82)°, and had significant meaning compared with preoperative (P<0.01). JOA score at the latest follow-up (14.6±1.1) was higher than that of before operation (8.2±1.9), and had siginificant differences (P<0.01).

CONCLUSIONSegmental anterior cervical decompression for the treatment of multilevel cervical myelopathy has a high clinical operability, and plays an important role in recovering cervical curvature and nerve function based on completely decompression.

Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Humans ; Male ; Middle Aged ; Spinal Fusion ; methods ; Spondylosis ; surgery

Objective To investigate the expression and significance of calcyclin binding protein (CacyBP)in the brain of rat model of traumatic brain injury(TBI).Methods Sixty 60 male SD rats were divided randomly into normal control group (n=10) and TBI group (n=50).The TBI model was created by using lateral head rotation device and subdivided into 6 h,24 h,72 h,7 d and 14 d group (10 rats per group).The expression and distribution of CacyBP in the rat brain was investigated immunohistochemically.The presence of the brown stained particles was considered aspositiveand lack of the stained particles agnegative. Results CacyBP was mainly distributed in the hippocampus,dentate gyrus and cortical neuron cytoplasm.Compared with the high level expression of CacyBP in the normal control group,the expression of CacyBP was decreased to the lowest in the rat brain at 6 h post TBI (P<0.01),became stronger gradually at 24 hours and recovered to normal at day 14,with no statistical difference compared with normal control group (P>0.05). Conclusion The lowest level expression of CacyBP after TBI indicates that CacyBP may play an important role in development of brain injury under effect of difierent mechanisms.

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

Objective To compare the effects of UVA irradiation on cell morphology,quantity and expression of inducible nitric oxide synthase (iNOS) mRNA and protein in human fibroblasts versus a kerati- nocyte cell line HaCaT.Methods Human fibroblasts and HaCaT cells were cultured and irradiated by 5 J/cm~2 UVA.Then,at 24,48 and 72 h after the stimulation,the cell morphology was observed under an in- verted microscope,and iNOS mRNA and protein were measured by RT-PCR and immunohistochemistry method,respectively.Results After the irradiation,human fibroblasts showed shrinkage at the three time points,the quantities of the cells began to decrease significantly at 24 h (P

Objective To investigate whether the donor's glomerular filtration rate (GFR) to recipient's weight ratio (Dg/Rw) is a useful tool to predict early clinical outcome in living-related do-nor transplantation.Methods A total number of 108 living donor transplant recipients in the Chinese Military 309th Hospital from Jan.2014 to July 2015 were enrolled in this study.The patients who had multi-organ transplantation or developed grafts rejection,delayed graft function,hydronephrosis or renal vascular stenosis were excluded.The 90 qualified recipients were divided into G1 group (Dg/Rw ≤0.81),G2 group (Dg/Rw 0.81～1.11),and G3 group (Dg/Rw≥1.12).We respectively analyzed the relationship between recipient's serum creatinine Scr and Dg/Rw at 3-,7-,30-day and 1 year after transplantation.Results Scr at 3-,7-,30-day and 1 year after transplantation had linear correlation with Dg/Rw.As compared with G1 and G2 groups,Scr level was significantly reduced in G3 group at different time points (P＜0.05).Conclusion Dg/Rw has a negative relationship with Scr level after renal transplantation.Pre-transplant Dg/Rw is a potential index to predict the early clinical outcome in living-related donor transplantation.

Objective To study the expression of basic fibroblast growth factor (bFGF),thrombospondin-1 (TSP-1) and the mi- crovessel density (MVD) and their relationship in esophageal squamous cell carcinoma.Methods Forty-six specimens of resected, paraffin embedded esophageal specious of squamous cell carcinoma were examined by immunohistochemical technique for the MVD and expression of bFGF and TSP-1.Results Of the 46 patients,the positive rate of bFGF and TSP-1 were 60.9% (28/46) and 37.0% (17/46),respectively.The mean value of MVD was (56.59?28.97)/mm~2.The depth of invasion had a significantly positive corre- lation with positive expression of bFGF and TSP-1.TNM stage had a significantly positive correlation with positive expression of TSP- 1.The value of MVD was closely associated with histological grade,lymph node metastasis and the expression of bFGF.Conclusion bFGF,TSP-1 and tumor microvessel play important roles in development and progression of esophageal squamous cell carcinoma. bFGF is a important regulator of tumor angiogenesis.

OBJECTIVE To map a comprehensive metabolic pathway of herbacetin in rats, specifically, to elucidate the biotransformation of herbacetin in vivo and to simultaneously monitor the pharmacokinetic process of both parent drug and its major metabolites. METHODS liquid chromatography/ion trap mass spectrometry (LC/MSn) and ultra-liquid chromatography coupled with mass spectrometry (UPLC/MS) were combined in the current study for qualitative and quantitative determinations of herbacetin and its metabolites in bile, urine and feces after both oral and intravenous administration of herbacetin to rats. Enzyme kinetic studies on the intestinal and hepatic metabolism of herbacetin were further conducted to elucidate metabolic profiles of herbacetin in rat tissues and organs. Additionally, plasma concentration profiles of herbacetin and its metabolites in rats were obtained to characterize the overall pharmacokinetic behavior of herbacetin. RESULTS It was found that herbacetin was excreted primarily from rat urine in the form of glucuronide-conjugations. Subsequent in vitro enzyme kinetic studies and in vivo pharmacokinetic investigations suggested an extensive hepatic metabolism of herbacetin and the high exposure of herbacetin- glucuronides in systemic circulation. The clearance, half- life and bioavailability of herbacetin in rats were determined as (16.4±1.92)mL·kg-1·min-1, (11.9±2.7)min, and 1.32%, respectively. On basis of these findings, a comprehensive metabolic pathway of herbacetin in rats was composed. In addition, a physiology based pharmacokinetic (PBPK) model was successfully developed with the aid of the GastroPlus to simulate the pharmacokinetic process of herbacetin in rats. Application of the PBPK modeling can provide a useful starting point to understand and extrapolate pharmacokinetic parameters among different species, populations, and disease states. CONCLUSION After oral administration, herbacetin was subjected to colonic degradation and extensive first pass metabolism, with glucuronidation as its dominating in vivo metabolic pathway.