Objective: To investigate the molecular mechanism of down-regulation of monocarboxylic acid transporter 1 (MCT1) on the proliferation inhibition of glioma cell. Methods: siMCT1, siMCT4 and negative control siRNA were transfected into glioma cell lines including U-251 and U-87. The proliferation activities of U-251 and U-87 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay and clonogenic assay. Glucose consumption and lactic acid efflux of U-251 and U-87 cells were determined by spectrophotometry.Western blot was used to detect the expressions of MCT1, MCT4, human glucose transporter 1 (GLUT1), GLUT4, tuberous sclerosis associated protein (TSC2), p-TSC2, 4E binding protein 1 (4EBP1), p-4EBP1, ribosomal S6 protein kinase (S6) and p-S6 protein in U-251 and U-87 cells. Results: Compared with negative control group, siMCT1 and siMCT4 significantly inhibited the expressions of MCT1 and MCT4 protein in U-251 and U-87 cells (both P<0.05). However, only knockdown of MCT1, the proliferation activities of U-251 and U-87 cells significantly decreased (P<0.05). The clone formation rates of U-251 and U-87 cells decreased to (55.20±3.27)% and (68.33±4.58) %, respectively (P<0.05). The glucose consumption of U-251 and U-87 cells in the negative control group at 72 hours were (82.65±6.66) pmol/L and (63.33±5.27) pmol/L, respectively, significantly higher than (31.70±3.17) pmol/L and (26.41±3.19) pmol/L of the siMCT1 transfected group (P<0.05). The extracellular lactate flow of U-251 and U-87 cells in negative control group at 72 h were (155.49±8.15) mmol/L and (135.37±8.21) mmol/L, respectively, significantly higher than (42.69±4.66) mmol/L and (38.91±4.83) mmol/L of the siMCT1 transfected group (P<0.05). Western blot analysis showed that knockdown of MCT1 significantly decreased the protein levels of GLUT1 p-TSC2, p-4EBP1 and p-S6 in U-251 and U-87 cells. Conclusions Downregulation of MCT1 expression can inhibit the proliferation of glioma cells. Deletion of MCT1 inhibits the glycolysis and metabolism of glioma cells through regulating the mTOR signaling pathway.

Objective To investigate clinical efficacy of transforaminal approach debridement with fusion,thoracolumbar single segment of Brucella spondylitis pedicle screw fixation (TLIF surgery).Methods We analyzed retrospectively the clinical data of 28 patients with Brucella spondylitis treated in our department between January 2009 and January 2014 with TLIF surgery (Group A) and internal fixation with a simple posterior anterior interbody disease debridement,autogenous bone graft (Group B).The two groups were compared in operation time,blood loss,postoperative ambulation time,hospitalization days,erythrocyte sedimentation rate (ESR),Creactive protein (CRP),American Spinal Injury Association (ASIA) classification,visual analogue scale (VAS),Oswestry Disability Index (ODI),Cobb angle of vertebral bone graft healing,and complications.Results All the patients were followed up for an average of 20.2 months (18 to 27 months).They were all cured.Compared with those in Group B,patients in Group A had shorter operation time (164.60±59.19)min,significantly reduced blood loss (346.00±108.90)mL and complications (1 case);significantly shorter postoperative ambulation time (3.36±1.11 days),hospitalization days (17.36 ± 4.19) days and duration (13.16 ± 3.94) months (P ＜ 0.05).The two groups did not significantly differ in VAS scores,ODI,ESR CRP,or Cobb angle (P＞0.05).Conclusion On the basis of norms of anti-drug treatment for brucellosis,TLIF surgery on Brucella spondylitis has the advantages including less trauma,shorter operation time,easier operation,less bleeding,earlier postoperative ambulation,and lower complication rate.

Objective To discuss clinical significance on early diagnosis of brain injury in premature infants with multiple sequence joint inspection of magnetic resonance imaging (MRI).Methods The brain MRI findings of 160 premature infants treated by Neonatal Intensive Care Unit were analyzed retrospectively.Results In 160 premature infants,brain injury occurred in 76 cases,the incidence of brain injury was 47.5％.Ischemic lesions were seen more in brain injury in premature infants,cerebral white matter injury was the most common,especially periventricular leukomalacia.Ischemic brain injury performed patchy or large sheet increased signal intensity on T1-weighted images(T1 WI),decreased signal intensity on T2-weighted images (T2WI) and obviously increased signal intensity on diffusion weighted imaging (DWI) in half egg circle center and around the lateral ventricle.Periventricular leukomalacia performed patchy decreased signal intensity on T1WI,increased signal intensity on T2WI and decreased signal intensity on DWI.Periventricular-intraventricular hemorrhage was seen more in hemorrhagic lesions.Hemorrhage stove was performed different signal because of different bleeding time.MRI performance in acute phase was iso-signal or slightly decreased signal intensity on T1WI,increased signal intensity on T2WI,increased signal intensity on T1WI,slightly decreased signal intensity on T2WI in early subacute,increased signal intensity on T1 WI and T2WI in late subacute and obviously decreased signal intensity on magnetic sensitive weighted imaging.The detection rate of ischemic lesions by DWI was higher than the conventional MRI,and DWI could show cerebral white matter damage of premature infants much earlier than the conventional MRI.The detection rate of hemorrhage stove by susceptibility weighted imagingc (SWI) was higher than the conventional MRI (x2 =23.78,P ＜ 0.05),and SWI could show hemorrhagic lesions much earlier than conventional MRI (x2 =27.02,P ＜ 0.05).Conclusions MRI,especially combined multiple sequence checking,could provide accurate imaging evidence for the early diagnosis of brain injury in premature infants.

Objective To explore the relationship between hypoxia-inducible factor-1α (HIF-1α) expression and apoptosis in early brain injury models after subarachnoid hemorrhage (SAH).Methods Fifty-five adult male Sprague-Dawley rats were randomly assigned to five groups:sham-operated group,SAH 6 h,SAH 12 h,SAH 24 h and SAH 72 h groups (n=1 1).SAH in the later four groups was induced by modified monofilament puncture method.The rats were killed by cervical dislocation.HIF-1α expression was assessed by immunofluorescence staining.TUNEL was adopted to detect brain apoptotic cells.Immunofluorescence double staining was used to identify cell types with positive HIF-1α expression.Pearson correlation analysis was employed to analyze the relationship between HIF-1 expression percentage and TUNEL positive rate.Results As compared with those in the sham-operated group,the HIF-1 expression percentage and TUNEL positive rate in the four SAH groups was significantly higher (P＜0.05).Immunofluorescence double staining showed that neuron-specific nuclear protein staining cells were coincided with most HIF-1 positive cells,while only a few HIF-1α positive cells were coincided with glial fibrillary acidic protein staining cells.A significant positive correlation was noted between HIF-1 α expression percentage and TUNEL positive rate following SAH (r=0.737,P=0.001).Conclusion HIF-1α high expression after SAH early promotes neuronal cell apoptosis,indicating HIF-1 a might participate in the pathological progression of early brain injury after SAH.

Objective To explore the compare of complete and incomplete radical debridement for thoracolumbar spinal tuberculosis.Methods Data of 296 patients with spinal tuberculosis from January 2000 to January 2011 were retrospectively analyzed.All patients were divided into two groups according to completeness of debridement:complete debridement group (group A) and incomplete debridement group (group B).There were 162 cases in group A including 86 males and 76 females,with an average age of 38.74± 17.26 years.There were 134 cases in group B including 73 males and 61 females,with an average age of 35.64± 18.21 years.All paticnts had undergone anterior debridement,focal graft implantation,anterior or posterior deformity correction,and internal fixation.Regular follow-up was required in the two groups.Results Residual sclerotic walls (36.54％),multipie cavities (34.62％),affected bony bridges (13.46％),sequestmm (3.37％),abscess (7.21％) and other lesionses (4.81％) were found in the group B.The first three factors were made up 84.62％ of the total.The mean follow-up time was 76.13±8.32 months in the group A and 79.24±5.49 months in the group B.The symptoms,C-reactive protein and erythrocyte sedimentation rate were improved more obviously in group A than those in group B.Six months after operation,tuberculosis healing rate in group A and group B was 29.01％ (47 patients) and 4.48％ (6 patients),respectively.The mean healing time was 4.36± 1.27 months in the group A and 9.15±2.53 months in the group B,with significant differences.The mean the time of chemotherapy was 5.21± 1.38 months in the group A and (10.45±2.15) months in the group B,with significant differences.Reoperation rate in group A and group B was 0.62％ (1/162) and 4.48％ (6/134),respectively.Conclusion Sclerotic bone,multiple cavities,and bony bridges are parts of foci in spinal tuberculosis.Clearing tuberculous foci with sclerotic bone,multiple cavities,and bony bridges can increase the curative effect,shorten the time of chemotherapy and reduce the side effects of drug,thus early resumption can be achieved.

BACKGROUND:Radiotherapy alone is not suitable for tumor-caused vertebral fractures and neurological dysfunction. In recent years, 125I radiation particles have been widely used in a variety of primary or secondary tumors and achieved good results. Percutaneous kyphoplasty can restore vertebral height efficiently, remodel spinal stability, and relieve pain.
OBJECTIVE:To evaluate safety and effectiveness of percutaneous kyphoplasty combined with 125I in patients with metastatic spinal tumors.
METHODS:A retrospective study was conducted to review 30 cases of metastatic spinal tumors undergoing percutaneous kyphoplasty combined with 125I from March 2011 to July 2012. Symptoms, signs, and imaging findings were col ected and analyzed. Al the patients had a refractoriness back pain. CT scan showed osteolytic changes in the vertebrae. The visual analogue scales, WHO standards for pain relief and Owestry disability index were recorded to analyze the clinical symptoms outcome and recovery of neurological function, and the change of height in abnormal vertebrae was measured. The fol ow-up time was 1 day, 1 month and 6 months postoperatively. RESULTS AND CONCLUSION:Operations in al the 30 patients were done successful y. Al patients got a conspicuous pain relief in 24 hours after operation, and nospinal injury or compression was found. There were significant differences in scores of visual analogue scales, pain levels, Owestry disability index, and the height of vertebral bodies before and after operation (P<0.05). During postoperative fol ow-up of 1 and 6 months, scores of visual analogue scales, pain levels, Owestry disability index, and the height of vertebral bodies showed no difference from those at 24 hours postoperatively (P>0.05). Bone cement leakage occurred in the anterior longitudinal ligament (n=2) and intervertebral space (n=2), and no serious complications occurred. Percutaneous kyphoplasty combined with 125I is a safe and effective way to treat metastatic spinal tumors, which can quickly ease the pain caused by spinal tumor, recover the abnormal vertebral height, reduce complications and improve life quality of patients.

Objective To study magnetic resonance imaging(MRI) features of dysembryoplastic neuroepithelial tumor(DNT) and to improve accurate diagnosis of DNT.Methods The MRI appearance and clinical features of 10 patients with DNT confirmed by surgery and pathology were analyzed retrospectively.Results In 10 cases,9 tumors located in supratentorial hemisphere cortex,3 tumors located in the temporal lobe,5 in the frontal lobe,1 in the parietal lobe,and 2 of them encroached the adjacent white matter.In 9 tumors located in supratentorial hemisphere cortex,8 cases had decreased signal intensity on T1-weighted MR images,1 case iso-decreased mixed signal intensity on T1-weighted MR images,and 9 cases increased signal intensity on T2-weighted images,9 cases slightly increased signal intensity on fluid attenuated inversion recovery weighted images.The manifestation of tumors was cystic or cystic partially oriented and was seen separate section intratumoral in some cases.Three cases appeared as hyperintense ring sign and internal septation,2 cases appeared as a triangle in shape,3 cases appeared as gyms-like shape,and 1 case as round shape,similar to cyst.Nine tumors had no significant mass effect and peritumoral edema.Enhanced MR imaging showed only 1 case with slight and heterogeneous enhancement,the rest 6 cases showed non enhancement.One case located in cerebellar hemisphere,and appeared cystic-solid mass,the solid part had decreased signal intensity on T1-weighted MR images,and increased signal intensity on T2-weighted images,the cystic part had decreased signal intensity on T1-weighted MR images,and increased signal intensity on T2-weighted images.On enhanced MR imaging,the wall-node obviously contrast enhancement,cyst wall slightly contrast enhancement,cystic part non enhancement.The tumor had peritumoral edema and mass effect.Ten cases had no hemorrhage and calcification.Conclusion The MRI appearance of DNT is characteristic and is helpful for the preoperative diagnosis of DNT.

BACKGROUND: Small conductance,Ca2+-activated potassium(SK)channel,presents in various cell types and plays a crucial role in action potential profile.However,coupling and modulation of calcium and associated molecules to SK2 channel remains unclear.OBJECTIVE: To construct the recombinants of pGBAT7 and target fragments of SK2 gene,so as to observe the coupling and regulation of SK2 channel gene to calcium and other molecules.METHODS: Three pairs of primers of the target fragments of SK2 gene were designed and synthesized based on the full-length sequences of SK2.After being identified,they were individually sub-cloned into the yeast expressive plasmid pGBKT7 to construct pGBKT7-SK2 vectors.The recombinant pGBKT7-SK2 vectors were transformed into yeast AH109 by electroporation,and their activation was tested.The recombinants were extracted from yeast AH109 and verified by electrophoresis and sequencing.RESULTS AND CONCLUSION: The target fragments of SK2 gene by PCR were 411,546 and 729 bp,respectively.Three sub-clones of pGBKT7-SK2 were successfully constructed.Electrophoresis and sequencing showed that the constructed sub-clones of pGBKT7-SK2 met the expected requirements.The recombinant pGBKT7-SK2 vectors transformed into the yeast could be activated.The successful construction of the sub-clones of SK2 gene provides an important material basis for further study in the SK2 channel and function-associated molecules.

OBJECTIVE To investigate the prevalence of ?-lactamases in nosocomial infections of Enterobacter cloacae,study the different genotyping of ?-lactamases,and analyze its independence in drug resistance. METHODS AmpC ?-lactamases and ESBLs were detected by three-dimensional extract tests.PCR was used to determine the genotype of ?-lactamases and their resistance to 13 kinds of antibiotics was detected by Kirby-Bauer method. RESULTS The sensitivity test showed that they were resistant to the most antibiotics.Among the 80 ESBLs strains,29 strains carried ESBLs gene,3 strains carried two different gene.Such as:TEM existed in 6,SHV-Ⅰ existed in 2,CTX-M-2 existed in 5,CTX-M-3 existed in 8 and CTX-M-9 existed in 11 strains.Twenty-six strains carried ampC.The resistant rate of SSBLs-producing E.cloacae to 12 antibiotics except merapenem was 66.7-100.0%.SSBLs strains of E.cloacae were higher than AmpC ?-lactamases in resistance to levofloxacin,cefepime,aztronem and trimethoprim sulpfamethoxazole with significant difference(P

BACKGROUND: Elimination of antigenic substances from natural extracellular matrix with the integrity of the tissue structure retained renders the matrix to possess better biocompatibility and provides a cell culture environment close to conditions of the internal environment. Such materials are the primary choice for cell culture scaffold in tissue engineering.OBJECTIVE: To prepare human articular cartilage acellular matrix so as to provide a methodological basis for further study of articular cartilage acellular matrix as cell scaffold materials.DESIGN: A single sample study of bone tissues.SETTING: The experiment was performed in Institute of Orthopedics, General Hospital of PLA, between January and May in 2004. The specimens were obtained from patients requiring joint replacement for femoral neck fracture.MATERIAIS: The experiment was conducted in the Department of Orthopedics, General Hospital of PLA from January to May in 2004. Human articular cartilage specimens were obtained from the femoral head of patients with total hip arthroplasty for femoral neck fracture.METHODS: Totally 10 specimens of fresh articular cartilage(3.5 mm × 4. 5 mm × 2.0 mm) were obtained and freeze-dried for 12 hours. Cartilage acellular matrix was prepared using Triton X-100, Dnase and Rnase and identified by means of hematoxylin-eosin(HE) and safranine O staining and immunohistochemical staining for cartilage proteoglycan.MAIN OUTCOME MEASURES: Histological observation of the articular cartilage acellular matrix and immunohistochemical staining of cartilage proteoglycan.RESULTS: HE and safranine O staining both showed no cellular structure in the matrix with only recesses left by the removed cells. Immunohistochemical staining for cartilage proteoglycan yielded positive results, suggesting the presence of cartilage proteoglycan in the acellular matrix.CONCLUSION: Human articular cartilage acellular matrix can be prepared using the modified four-step procedures with detergent and enzymatic extraction with lyophilization, and the preserved cartilage proteoglycan in the material may retain good pressure resistance.