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.

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 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

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

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.

OBJECTIVE To investigate the effect of hypoxia on the pharmacokinetic process of salidrosidein rats and to explore its underlying mechanisms. METHODS The Caco-2 cell monolayerwas exposed to 1% oxygen (O2) concentration for 24 h to build the hypoxiccell model. The transportation mode of salidroside was investigated with the aid of this hypoxia model by detecting the apparent permeability coefficient(Papp). Healthy Sprague Dawley (SD) rats were exposed to 9% O2 for 72 h for the construction of hypoxic rat model. Liver sample was subsequently collected from the hypoxic rats with an aim to identify enzymes responsible for salidroside metabolism. The expression levels of sali?droside-transporting and salidroside-metabolizing enzymes, including Sodium-dependent glucose cotrans?porters (SGLT1), β-glucosidase (GBA3)and sulfotransferase (SULT2A1), were thereafter detected by RT-PCR and Western blot. The metabolic activity of GBA3 and SULT2A1 was monitored by rat liver microsome incubation.In addition, the renal function of rats under hypoxia was assessed by detecting concentrations of blood urea nitrogen and creatinine. RESULTS The AUC and t1/2 values of salidroside in hypoxic rats were more than doubled, while the in vivo clearance was significantly reduced. Mechanistic study demonstrated that the PappA- B/PappB- A eualsto 10.3, indicating the potential active transport of salidrosile. The expression of SGLT1 and GBA3 was significantly decreased, which indicated a reduced metabolism of salidroside under hypoxia. Moreover, rat under hypoxia was found to suffer from renal dysfunction, with an abnormal value of blood urea nitrogen. CONCLUSION Due to the reduced metabolism and the abnormal renal function under hypoxia, the systemic exposure of salidroside in rats was signifi?cantly enhanced.

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 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 observe the changes in percentages and subsets of invariant nature kiler T (iNKT) cells in adipose and related tissues at different stages of obesity, and analyze the role of iNKT cells during chronic inflammation in adipose tissues in a mouse model of obesity established with high-fat diet.Methods:Changes in mouse body weight, mental state, glucose tolerance and insulin tolerance were recorded. Hematoxylin and eosin (HE) staining was used to observe pathological changes in adipose tissues. Flow cytometry was performed to detect the percentages and subsets of iNKT cells as well as the percentages and subtypes of macrophages. The levels of cytokines in serum samples and the culture supernatants of lymphocytes in adipose tissues were detected with CBA. The expression of related proteins in adipose tissues was detected by Western blot.Results:(1) The volume of adipose cells increased significantly after four weeks of high-fat feeding, but the infiltration of inflammatory cells was not obvious. Significantly increased infiltration of inflammatory cells was observed after 12 weeks of high-fat feeding. (2) High-fat feeding could reduce the percentage of iNKT cells, increase the proportion of iNKT1 subgroup and decrease the proportion of iNKT10 subgroup in adipose tissues. The proportion of iNKT1 subgroup in thymus increased, but that of iNKT2 subgroup decreased. The percentage of macrophages and the proportion of M1 subgroup in adipose tissues increased, while the proportion of M2 subgroup decreased, which were more obvious after 12 weeks of high-fat feeding. (3) High-fat feeding resulted in decreased expression of E4BP4 and arginase-1 (Arg-1) in adipose tissues and increased expression of inducible nitric oxide synthase (iNOS). (4) High-fat feeding significantly increased the pro-inflammatory cytokines and decreased the anti-inflammatory cytokines in mouse serum and culture supernatants of lymphocytes in adipose tissues with more significant changes observed after 12 weeks of high-fat feeding.Conclusions:Increased iNKT1 and decreased iNKT10 in obese adipose tissues might be closely related to the increased M1 polarization and the imbalance of iNKT subsets might be involved in the progression of chronic inflammation in obese adipose tissues.