Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Low Back Pain ; etiology ; Male ; Spinal Puncture ; adverse effects

To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona ; microbiology ; Ascomycota ; chemistry ; genetics ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; metabolism ; Endophytes ; chemistry ; genetics ; isolation & purification ; metabolism ; Lactones ; chemistry ; isolation & purification ; metabolism ; Mass Spectrometry ; Molecular Structure

The growth curve of single cell organisms in batch cultivation could divide into 6 phases, lag phase, acceleration phase, log phase, deceleration phase, stationary phase, and death phase, based on specific growth rate during cultivation process. There were significantly differences between deceleration phase and the other phases in cell growth, substrate consumption, product formation, and genes express profile. The deceleration phase was highly important to fermentation process. However, cognizing and teaching to the deceleration phase had been considerably weakened since a long period. So it should be strengthened.

OBJECTIVENeuroblastoma (NB) is a pediatric solid tumor derived from neural crest precursor cells. It is resistant to current therapeutic protocols, including high dose chemotherapy. The mechanisms of chemoresistance are very complex. The recent studies have shown that the levels of tyrosine kinase receptor B (TrkB) and brain-derived neurotrophic factor (BDNF) are high in NB tumors with poor prognosis. The aim of this research was to explore the effects of TrkB and BDNF levels on the chemotherapeutic sensitivity in neuroblastoma by using the NB cell line SH-SY5Y in vitro.

METHODSThe expression of TrkB protein was detected with Western-blot after the treatment with different concentrations of all trans-retinoic acid (ATRA). Cell survival rate was analyzed using MTT. Apoptosis was detected using flow cytometry (FCM) and a transmission electron microscope (TEM).

RESULTS(1) The expression of TrkB protein was undetectable in SY5Y. It was positive, however, after the treatment with ATRA (1, 10, 100 nmol/L) for five days. The level of TrkB protein was increased with adding of ATRA at different concentrations. (2) The difference of the survival and apoptotic rate between the BDNF (10 ng/ml) + ATRA (10 nmol/L) + cisplatin (CP, 5 microg/ml) group (survival rate 46.51% +/- 13.44%, apoptosis rate 11.79% +/- 1.53%) and the CP alone group (survival rate 38.51% +/- 9.66%, apoptosis rate 14.95% +/- 2.06%) was not statistically significant (P > 0.05). The survival rate of the BDNF (50 ng/ml and 100 ng/ml) + ATRA (10 nmol/L) + CP (5 microg/ml) group (66.85% +/- 18.39%, 94.30% +/- 10.71%) was greatly higher than CP alone group (P < 0.05, P < 0.01), whereas the apoptotic rate (9.36% +/- 1.03%, 5.20% +/- 1.99%) was significantly lower than that of the CP alone group (P < 0.01, P < 0.01). The survival rates of BDNF (100 ng/ml) + ATRA (10 nmol/L) + CP (5 microg/ml) group were higher than those of BDNF (50 ng/ml) + ATRA (10 nmol/L) + CP (5 microg/ml) group (P < 0.01), whereas the apoptotic rates were lower than those of BDNF (50 ng/ml) + ATRA (10 nmol/L) + CP (5 microg/ml) group (P < 0.05). There were no significant difference between the ATRA (1 nmol/L) + BDNF (50 ng/ml) + CP group (survival rate 45.33% +/- 11.83%, apoptosis rate 12.48% +/- 2.48%) and the CP alone group in the survival and apoptotic rates (P > 0.05). The survival rates of the ATRA (10 nmol/L, 100 nmol/L) + BDNF (50 ng/ml) + CP (61.62% +/- 18.53%, 105.02% +/- 5.55%) group were greatly higher than those of the CP alone group (P < 0.05, P < 0.01), whereas the apoptotic rate (9.36% +/- 1.03%, 5.05% +/- 1.88%) was significantly lower than that of the CDDP alone group (P < 0.05, P < 0.01). The survival rates of the ATRA (100 nmol/L) + BDNF (50 ng/ml) + CP group were higher than those of the ATRA (10 nmol/L) + BDNF (50 ng/ml) + CP group (P < 0.01), whereas the apoptotic rates were lower than the ATRA (10 nmol/L) + BDNF (50 ng/ml) + CP group (P < 0.01). (3) Some of the cells showed apoptotic changes in the CP alone group, whereas the intranuclear chromoplasma was well-distributed, the nuclear membrane was clear, and mitochondria, ribosome and solvent were present in the ATRA (10 nmol/L) + BDNF (50 ng/ml) + CP group.

CONCLUSIONSThe sensitivity of SY5Y to CP was affected by TrkB and BDNF. The higher the level of TrkB and BDNF was, the lower the sensitivity of SY5Y to CP.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Brain-Derived Neurotrophic Factor ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; drug effects ; Flow Cytometry ; Humans ; Microscopy, Electron, Transmission ; Neuroblastoma ; drug therapy ; metabolism ; pathology ; ultrastructure ; Receptor, trkB ; metabolism ; Tetrazolium Salts ; chemistry ; Thiazoles ; chemistry ; Tretinoin ; administration & dosage ; pharmacology

OBJECTIVEAn electric monitor of bruxism had been invented in order to evaluate the curative effect of cuspid occlusal upheaving splint and stabilization splint.

METHODS20 patients with bruxism were randomly divided into two groups. A cuspid occlusal upheaving splint or a stabilization splint was fabricated respectively for patients. The vertical dimension for each splint was 0.5 mm lower than mandibular postural position. Sleeping time, bruxist time and times of bruxism were recorded with bruxism monitor that was invented for studying bruxing.

RESULTSThe bruxist time and the times of bruxism were decreased obviously in patients with cuspid occlusal upheaving splint, while no significant difference was shown before and after using the stabilization splint.

CONCLUSIONSThe bruxism monitor can automatically measure and record the data of bruxism with splint, which is valuable for clinic. The curative effect of cuspid occlusal upheaving splint is better than that of stabilization splint for treating bruxism.

Adult ; Bruxism ; physiopathology ; therapy ; Electromyography ; Female ; Humans ; Male ; Masticatory Muscles ; physiopathology ; Occlusal Splints

OBJECTIVETo evaluate the clinical outcome of arthroscopically assisted combined anterior and posterior cruciate ligament (ACL/PCL) reconstructions using Achilles tendon-bone allografts.

METHODSAssociated meniscus injuries were treated according to established methods prior to ligament reconstructions during arthroscopic surgery. Thirty Achilles tendon-bone allografts were used to reconstruct torn ACL and PCL in 15 knees. At postoperative follow-up, all knees were graded using the modified IKDC and the Lysholm scoring systems just as done preoperatively.

RESULTSwere analyzed compared with the contralateral healthy knees. Results: Eleven men and 4 women with a minimum of 3-year follow-up (mean 38 months) were included in the study. Preoperatively, the group ratings by the modified IKDC standards were all severely abnormal. Twelve bicruciate reconstructions were performed in subacute or chronic stage (larger than 3-8 weeks), 3 for acute ligamentous deficiencies (less than or equal to 3 weeks). The noticeable early complication was transitory local fever combined with joint effusion in one case. At postoperative follow-up, 9 knees were normal, 5 nearly normal and 1 abnormal. On Lysholm score the difference was statistically significant (t- test, P less than 0.001) before and after operation.

CONCLUSIONSAchilles tendon-bone allograft offers an alternative for simultaneous arthroscopic ACL/PCL reconstructions. However, further investigation is needed to eradicate its potential immunogenicity for better use.

Achilles Tendon ; transplantation ; Anterior Cruciate Ligament ; surgery ; Arthroscopy ; methods ; Bone Transplantation ; methods ; Female ; Humans ; Knee Injuries ; surgery ; Male ; Posterior Cruciate Ligament ; surgery ; Range of Motion, Articular ; Reconstructive Surgical Procedures ; methods ; Transplantation, Homologous

OBJECTIVETo selectively knockdown the expression of Angiotensin II receptor subtype 1a (AT1aR) in rat vascular smooth muscle cells (VSMCs) by RNA interference and the sequential effects on cellular viability and proliferation.

METHODSThe primary cultured rat aortic VSMCs were transfected by plasmids pAT1a-shRNA1 and pAT1a-shRNA2, each carrying an U6 promoter and an AT1a-specific shRNA-coding template sequence, or by a control plasmid pGenesil-Control (pCon) carrying a nonspecific shRNA-coding sequence. The mRNA and protein expressions of AT1a, AT2 were analyzed by semi-quantified RT-PCR and Western blot, respectively and normalized to the internal control gene beta-actin. Cellular viability and proliferation were determined with methylthiazoletetrazolium (MTT) assay.

RESULTSAT1a mRNA and protein were reduced by 82% and 69% by pAT1a-shRNA1, 77% and 56% by pAT1a-shRNA2, respectively while no change was found in pCon treated VSMCs. AT2 receptor level in VSMCs remains unchanged after various treatments. The A(490nm) values obtained by MTT measurements were similar among groups in the absence of Ang II but decreased significantly in pAT1a-shRNA1 and pAT1a-shRNA2 treated VSMCs in the presence of Ang II.

CONCLUSIONRNA interference can selectively knockdown AT1a expression in cultured VSMCs and attenuate the Ang II induced cell proliferation. Future studies are warranted to explore the potential role of RNA interference on AT1 function and as a new gene therapy tool for cardiovascular diseases.

Animals ; Cells, Cultured ; Gene Knockdown Techniques ; Male ; Muscle, Smooth, Vascular ; metabolism ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Receptors, Angiotensin ; metabolism ; Transfection

OBJECTIVETo investigate the short-term efficacy and safety of triple semicircular canal occlusion (TSCO) in the treatment of intractable Meniere's disease (MD), so as to provide an alternative surgical procedure for treating MD.

METHODSSeventeen patients, who had received standardized conservative treatment for at least one year with poor effect, underwent TSCO were retrospectively analyzed. Vertigo control and auditory function were evaluated. Pure tone audiometry, caloric test, and vestibular evoked myogenic potential (VEMP) were performed for evaluation of audiological and vestibular function. Postoperative follow-up period was 6 - 13 months, with an average of ten months.

RESULTSAccording to the preoperative staging of hearing, among the 17 patients, there were 2 cases in stage II (with an average hearing threshold of 25 - 40 dBHL) and 15 in stage III (41 - 70 dBHL). No vertigo was found during the follow-up period, with 100% control rate of vertigo. During the same period, we had performed endolymphatic sac decompression operation in 25 MD patients. The control rate of vertigo was 72.0%. The vertigo control rate of TSCO was significantly higher than that of endolymphatic sac decompression operation (χ(2) = 3.87, P < 0.05). Three months after surgery, 12 patients showed no significant change in comparison to primary status, 5 patients presented with an mild increase in the average hearing threshold of less than 20 dBHL, with 29.4% of hearing loss rate. Post-operatively, all patients suffered from temporary vertigo and balance disorders. Vertigo was disappeared in all patients within 3 days, while, balance disorders were disappeared in 10 patients within 1 - 2 weeks after surgery, and in another 7 patients within 2 months, with an average recovery time of 12.6 days. Three months after treatment, loss of semicircular canal function by caloric test was found in the operation side of all patients and no change in VEMP test was noted. All patients had no facial paralysis, cerebrospinal fluid leakage, and other complications.

CONCLUSIONSTSCO, which can reduce vertiginous symptoms in patients with intractable MD, represents an effective and safe therapy for this disorder. TSCO is expected to be used as an alternative procedure for the treatment of MD in selected patients suffering from moderate to severe hearing loss.

Adult ; Aged ; Female ; Humans ; Male ; Meniere Disease ; surgery ; Middle Aged ; Otologic Surgical Procedures ; methods ; Retrospective Studies ; Semicircular Canals ; surgery ; Treatment Outcome

OBJECTIVETo explore the relationship between epidermal growth factor receptor (EGFR) gene expression and radiosensitivity of non-small-cell lung cancer (NSCLC) cells.

METHODSEGFR sequence-specific double-stranded RNA (dsRNA-EGFR) was chemically synthesized. NSCLC cell line SPC-A1 was transfected with dsRNA-EGFR formulated with Lipofectamine 2000. Western blot and real-time PCR were used to determine the EGFR mRNA and protein expression, respectively. Colony inhibition test was adopted to observe the radiosensitizing effect. To establish the nude mouse tumor models, calculate the tumor growth inhibition rate and make the tumor growth curve by measuring its size and weight.

RESULTSEGFR mRNA levels were 1.51 ± 0.22, 1.38 ± 0.15 and 0.45 ± 0.11 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 482.7, P < 0.01). The contents of EGFR protein were 2340.87 ± 10.99, 2231.85 ± 35.66 and 832.03 ± 39.13 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 263.3, P < 0.05). Compared with the control group, dsRNA-EGFR sequence specifically decreased the expressions of EGFR mRNA by 70.2% and EGFR protein by 64.5%. The colony inhibition rates of the control group, dsRNA-unrelated combined with radiotherapy group and dsRNA-EGFR combined with radiotherapy group were 9.3%, 12.5% and 65.5%, and the tumor growth inhibition rates were 21.3%, 24.4% and 64.2%, respectively. The combination of dsRNA-EGFR and radiotherapy significantly inhibited the tumor growth in vitro and in vivo.

CONCLUSIONSDsRNA-EGFR shows an apparent inhibitory effect on the expression of EGFR mRNA and protein of NSCLC cells, effectively inhibit the tumor growth in vivo, and enhance the radiosensitivity of NSCLC.

Adenocarcinoma ; metabolism ; pathology ; radiotherapy ; Animals ; Cell Line, Tumor ; Cell Proliferation ; radiation effects ; Humans ; Lung Neoplasms ; metabolism ; pathology ; radiotherapy ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Double-Stranded ; genetics ; RNA, Messenger ; metabolism ; Radiation Tolerance ; Random Allocation ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Transfection ; Tumor Burden ; radiation effects

Naringenin (1) was transformed to three metabolites (2-4) by Mucor sp. Based on LCMS(n)-IT-TOF and NMR spectroscopic data, 2-4 were identified as naringenin-7-O-sulphate, naringenin-4'-O-sulphate, and naringenin-5-O-sulphate, respectively. These results might provide hints to the mammalian/human metabolism of naringenin.
Biotransformation ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Flavanones ; chemistry ; metabolism ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Mucor ; metabolism ; Sulfates ; metabolism