Humans ; Nasal Cavity ; pathology ; Nasal Obstruction ; diagnosis ; Nasal Polyps ; diagnosis ; Turbinates

OBJECTIVETo evaluate the short and long term results of discectomy for lumbar intervertebral disc herniation.

METHODSFrom 2000 to 2007, 400 patients (male 220 and female 180, the age was from 16 to 73 years old with an average of 42.3 years) with lumbar intervertebral disc herniation underwent discectomy by posterior mini-incision less than 5 cm and vertebrae plate was ectomized in 2 cm x 2 cm winder,and nerve root was compressied. The short and long term clinical result were analyzed with SPSS 10.0 software.

RESULTSThree hundred and eighty patients were followed up in the short term (less than 2 years after operation), 308 cases obtained excellent result, 48 good, fair 24, the excellent and good rate was 93.7%. Three hundred and forty-eight patients were followed up in the long term (more than 3 years after operation), 244 cases obtained excellent result, 48 good, fair 56,the excellent and good rate was 83.9%. There was significant difference in follow-up between the short and long term (P < 0.05).

CONCLUSIONThe clinical effect of discectomy for lumbar intervertebral disc herniation decreased with time lapse.

Adolescent ; Adult ; Aged ; Diskectomy ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; surgery ; therapy ; Lumbar Vertebrae ; pathology ; Male ; Middle Aged ; Time Factors ; Treatment Outcome

A novel white-rot fungus AH28-2,which was isolated from 224 fungi samples,ability to produce effectively laccase by induction.Several factors influencing laccase production were investigated.The optimum conditions were as follows:the 300mL Erlenmeyer flask containing 150mL of liquid medium was inoculated with 7.5mL of mycelial fragments and the medium was supplied with lignin at a concentration of 0.1%(initial pH8.5).The cultures was incubated at 28℃ on rotary shaker(150r/min) for 4～5 days.The maximum enzyme activity was 20184IU/L.

The production conditions of extracellular laccase from Armilliria mellea and the characteristics of the enzyme were studied. The experiment proved that initial pH5.5 of the culture medium and temperature at 25 degrees C were favorable for laccase synthesis. As carbon resources, cellobiose and raffinose were better in terms of productivity than maltose, sorbose and galactose. Organic nitrogen source was more suitable for Armilliria mellea to synthesize laccase than inorganic nitrogen source. Peat extract (PE) enhanced notably the yield of laccase; the maximal yield was 7 times as much as that of the control when PE concentration was 50%. Three isozymes were detected in culture supernatant named A, B and C respectively after their mobility on PAGE. After concentrated by (NH4)2SO4 precipitation, laccase A was further purified to homogeneity by preparative native PAGE and anion exchange column chromatography. The native enzyme was a single polypeptide with a molecular mass of approximately 59 kD estimated by SDS-PAGE, while 58 kD by gel filtration chromatography under non-denaturing conditions. Determined by IEF its isoelectric point was 4.0. The optimal pH value and temperature were 5.6 and 60 degrees C respectively in catalytic reaction of oxidizing guaiacol. At 60 degrees C and 65 degrees C, half-lives of laccase A were 45 min and 36.8 min, respectively. Enzyme activity was inhibited with 100 mmol/L Cl-, but was activated with 1 mmol/L SO4(2-). However, if the concentration of NaN3 was only 1 mmol/L, laccase A lost its activity completely. 10 mmol/L EDTA had no effect on laccase A, while 1 mmol/L Cu2+ could enhance its activity. Laccase A showed a good stability when the pH of the buffer varied from 5.2 to 7.2. Using guaiacol as the substrate, the Km was 1.026 mmol/L and the Vmax was 5 mumol/(min.mg); using ABTS instead, the Km was 0.22 mmol/L and Vmax was 69 mumol/(min.mg).
Agaricales ; drug effects ; enzymology ; growth & development ; Carbohydrates ; pharmacology ; Culture Media, Conditioned ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Hydrogen-Ion Concentration ; Laccase ; Oxidoreductases ; isolation & purification ; metabolism

OBJECTIVETo investigate the effects and clinical pregnancy outcomes of intracytoplasmic sperm insemination (ICSI) with microamount frozen-thawed sperm obtained by percutaneous epididymal sperm aspiration (PESA) or testicular sperm aspiration (TESA) in azoospermia patients.

METHODSWe divided 365 azoospermia patients treated by ICSI into an experimental group (n = 123) and a control group (n = 242) , the former with microamount frozen-thawed sperm, and the latter fresh sperm obtained by PESA or TESA. The rates of fertilization, good embryos, clinical pregnancy, miscarriage, ectopic pregnancy and multiple pregnancy were analyzed and compared between the two groups.

RESULTSWith PESA, the experimental group showed no statistically significant differences from the control group in the rates of fertilization (75.67% vs 76.49%), good embryos (64.96% vs 66.09%), clinical pregnancy (55.21% vs 57.22%), clinical miscarriage (13.21% vs 12.61%), ectopic pregnancy (3. 77% vs 5.41%) and multiple pregnancy (37.74% vs 37.84%) (P > 0.05); nor with TESA (74.41% vs 76.43%, 64.63% vs 66.35%, 46.81% vs 53.39%, 18.18% vs 14.55%, 4.55% vs 1.82%, 37.74% vs 37.84%, P > 0.05). The revival rate of the frozen-thawed sperm from PESA was 70.07%, not significantly different from that of TESA (62.67%) (P > 0.05).

CONCLUSIONICSI with frozen-thawed micro-amount sperm obtained by PESA or TESA is a safe, economic and effective method for the treatment of azoospermia. The techniques for reviving frozen sperm from PESA or TESA remain to be optimized, and whether these techniques may result in long-term genetic risks in the offspring deserves further investigation.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Retrieval

OBJECTIVETo investigate the correlative factors of non-surgical vertebral fractures after percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fractures(OVCFs).

METHODSFrom August 2009 to September 2011, 126 patients who underwent single-level PVP for OVCFs were included in this study. They were followed up with an average time of 13.6 months,divided into the refracture group and non-refracture group according to the onset of non-surgical vertebral fractures or not. In refracture group,there were 14 males and 18 females with an average age of (67.63+/-7.28) years(ranged, 54 to 82); and in non-refracture group,there were 40 males and 54 females with an average age of (66.26+/-6.79) years (ranged, 55 to 76). The refracture group wps divided again into adjacent vertebral fracture (AVF) group (7 males and 13 females) and remote vertebral fracture(RVF) group (4 males and 8 females). The age, sex, bone mineral density(BMD), injecting bone cement volume, the recovery rate of vertebral body height,kyphosis corrected degree were recorded and the correlative factors of non-surgical vertebral fractures were analyzed.

RESULTSThere was no statistically significant differences in age, sex, BMD, injecting bone cement volume and kyphosis corrected degree between refracture group and non-refracture group (P>0.05), and there was statistically significant difference in the recovery rate of vertebral body height (P<0.05). There was no statistically significant difference in BMD, kyphosis corrected degree between adjacent vertebral fracture group and non-refracture group (P>0.05); and there was statistically significant difference in injecting bone cement volume,recovery rate of vertebral body height(P<0.05). There was no statistically significant difference in BMD,injecting bone cement volume,recovery rate of vertebral body height, kyphosis corrected degree between remote vertebral fracture group and non-refracture group (P>0.05).

CONCLUSIONRecovery of vertebral body height may prefigure increasing risk of refracture in non-surgical vertebral body for the patient with OVCFs after PVP, and the adjacent vertebral fracture maybe concerned with injecting bone cement volume and recovery rate of vertebral body height.

Aged ; Aged, 80 and over ; Bone Density ; Female ; Fractures, Compression ; surgery ; Humans ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery ; Spinal Fractures ; etiology ; Vertebroplasty ; adverse effects

OBJECTIVESTo study the different expression of miRNA between pediatric and adult types of brainstem gliomas, and to provide the target miRNAs for explore the mechanism and miRNA interference of the malignant progression of pediatric BSG.

METHODSmiRNA expression profiles in orthotopic models which could simulate the BSG heterogeneity were examined by microarray and analyzed to obtain the aberrantly expressed miRNAs. The two types of human BSG tissue were utilized to verify the microarray data by qRT-PCR and in situ hybridization for the putative causative miRNAs.

RESULTSThere were 216 miRNAs detected in both the pediatric BSG group and the adult BSG group, 39 miRNAs to be differential expressed in the pediatric BSG group versus adult group, including 10 up-regulated and 29 down-regulated. qRT-PCR and in situ hybridization indicated good consistency with that of the microarray method.

CONCLUSIONSAberrantly expressed miRNA may serve as putative causative involvement of malignant progression of pediatric BSG, thereby might be potentially novel targets for therapy.

Adult ; Age Factors ; Animals ; Brain Stem ; Brain Stem Neoplasms ; metabolism ; Child ; Disease Models, Animal ; Female ; Gene Expression Profiling ; Glioma ; metabolism ; Humans ; In Situ Hybridization ; MicroRNAs ; metabolism ; Oligonucleotide Array Sequence Analysis ; Rats

OBJECTIVETo study the inhibitory effect of knocking down microRNA(miR)-221 and miR-222 on human glioma cell growth and its possible mechanism.

METHODSmiRNA-221/222 antisense oligonucleotides (antisense miR221/222) were transfected into human glioma U251 cells by lipofectamine. Northern blot analysis was conducted to detect the mRNA expression of miR-221/222 in the control and transfected cell groups. The proliferation activity of cells was determined by MTT assay. Cell invasion ability was examined by transwell assay, and cell cycle kinetics and apoptosis were detected with flow cytometry. The expression of relevant proteins was analyzed by Western blotting. The therapeutic efficacy of antisense miR221/222 on the growth of xenograft tumors in nude mice were also observed.

RESULTSIn the antisense miR-221/222-transfected cells, the expression of miR-221/222 was significantly reduced; the cell invasion ability was suppressed, cell cycle was blocked at G(0)/G(1) phase, and apoptotic cells were increased. The growth of xenograft tumors treated with antisense miR-221/222 was also inhibited. In antisense miR-221/222 treated tumor cells, the expression of bcl-2 was down-regulated while connexin43, p27, PUMA, caspase-3, PTEN, TIMP3 and Bax up-regulated, and p53 expression not changed.

CONCLUSIONThere is a significant inhibitory effect of antisense miR-221/222 on the growth of human glioma U251 cells. miR-221/222 may be considered as a candidate target for gene therapy of human gliomas.

Animals ; Apoptosis ; Base Sequence ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Genetic Therapy ; Glioma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; MicroRNAs ; biosynthesis ; genetics ; Molecular Sequence Data ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-3 ; metabolism ; Transfection

A laccase gene (lacD) from the basidiomycete Trametes sp. 420 was heterologously expressed in Pichia pastoris in two ways, resulting in two recombinant enzymes of rLacDx with native N-terminus and rLacDe with eight additional amino acid residues at N-terminus. The yields of rLacDx and rLacDe in shaken-flask cultures after an 18-day growth were 1.21 x 10(5) u/L and 7.38 x 10(4) u/L, respectively, as determined with 2,2'-azinobis(3-ethylbenzothia-zoline- 6-sulfonic acid) (ABTS) as substrate. The yield of rLacDx was further increased to 2.39 x 10(5) u/L under high-density fermentation while the production process was decreased to 7.5 days. In addition, rLacDx and rLacDe exhibited similar enzymatic characters in oxidizing substrate guaiacol, and were stable at 50 degrees C and at a pH range from 3 to 10. However, the specific activity of rLacDx (1761 u/mg) for ABTS was higher than that of rLacDe (1122 u/mg), and the apparent Km value of rLacDx (427 microM) was less than that of rLacDe (604 microM).
Cloning, Molecular ; Fermentation ; Isoenzymes ; biosynthesis ; genetics ; Laccase ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Trametes ; enzymology ; genetics

OBJECTIVETo study the effect of silencing Dicer by small interference RNA (siRNA) to suppress the global microRNA (miRNAs) expression on the biological characteristics of TJ905 glioblastoma cells.

METHODSThe silencing effect of RNA interference on Dicer expression was evaluated by reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis and immunofluorescence staining. The cell proliferation rate and cell cycle kinetics were detected by MTT assay and flow cytometry respectively, and the cell invasive ability was evaluated by transwell assay.

RESULTSThe siRNA targeting Dicer suppressed the expression of Dicer in TJ905 cells. Meanwhile, the proliferation activity and invasive ability were significantly enhanced in cells transfected with Dicer siRNA compared to those cells transfected with scrambled siRNA and the control cells.

CONCLUSIONSuppression of Dicer expression renders the glioma cells harboring more aggressive phenotype. This preliminary finding suggests that global lower expression of miRNAs may play an oncogenic role.

Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DEAD-box RNA Helicases ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Glioblastoma ; genetics ; metabolism ; physiopathology ; Humans ; RNA, Small Interfering ; genetics ; metabolism ; Ribonuclease III ; genetics ; metabolism