To investigate cytotoxic secondary metabolites of Micrococcus sp. R21, an actinomycete isolated from a deep-sea sediment (-6 310 m; 142 degrees 19. 9' E, 10 degrees 54. 6' N) of the Western Pacific Ocean, column chromatography was introduced over silica gel, ODS, and Sephadex LH-20. As a result, eight compounds were obtained. By mainly detailed analysis of the NMR data, their structures were elucidated as cyclo(4-hydroxy-L-Pro-L-leu) (1), cyclo(L-Pro-L-Gly) (2), cyclo( L-Pro-L-Ala) (3), cyclo( D-Pro-L-Leu) (4), N-β-acetyltryptamine (5), 2-hydroxybenzoic acid (6), and phenylacetic acid (7). Compound 1 exhibited weak cytotoxic activity against RAW264. 7 cells with IC50 value of 9.1 μmol x L(-1).
Animals ; Biological Factors ; chemistry ; isolation & purification ; metabolism ; pharmacology ; Cell Survival ; drug effects ; Macrophages ; cytology ; drug effects ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Mice ; Micrococcus ; chemistry ; genetics ; isolation & purification ; metabolism ; Molecular Structure ; Phylogeny ; RAW 264.7 Cells ; Seawater ; microbiology ; Secondary Metabolism

OBJECTIVETo explore the advantages and indications of the paraspinal approach by anatomical study and clinical application.

METHODSThe anatomical data and clinical practice of 27 cases were analyzed to explore the accurate approach between the paraspinal muscles and the structure of ambient tissues, as well as the results of clinical application of paraspinal approach. The operation time, blood loss, incision length, radiographic result (Cobb angle, height of anterior edge of the vertebrae) were compared with those in 24 cases treated by traditional approach.

RESULTSComplete exposure of the facets could be easily performed by identifying natural cleavage plane between the multifidus and longissimus muscles. The natural muscular cleavage was (1.47+/-0.23) cm lateral to the midline for females, and (1.64+/-0.35) cm for males at T(12) level. The distance was (3.3+/-0.6) cm lateral to the midline for females, and (3.7+/-1.0) cm for males at L(4) level. In paraspinal approach group, the operation time was (76.2+/-15.7) min, blood loss was (91.6+/-16.9) ml and incision length was (7.6+/-0.8) cm. In traditional approach group, the operation time was (121.4+/-19.6) min, blood loss was (218.7+/-32.3) ml and incision length was (17.4+/-2.1) cm. To compare paraspinal approach with traditional approach, the operation time, blood loss and incision length had statistical difference (P less than 0.05) and the radiographic result (Cobb angle, height of anterior edge of the vertebrae) had no statistical difference (P larger than 0.05).

CONCLUSIONSWhen the paraspinal approach is performed through natural cleavage plane between the multifidus and longissimus muscles, there are no wide muscular disinsertions, leaving the supraspinous and interspinous ligaments intact. The distance of natural cleavage to the midline is different at T(12) and L(4) planes. By this approach, the facet joints can be explored easily and completely, and a clear surgical field will be available for the placement of pedicle screws. As a minimally invasive approach, it can be widely used in thoracolumbar spine surgery.

Female ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery ; Time Factors

OBJECTIVETo observe the antagonist effect of Curcuma Aromatica (CA) on renal tubular epithelial-myofibroblast transdifferentiation (EMT) induced by transforming growth factor-beta1 (TGF-beta1).

METHODSNormal renal tubular epithelial NRK-52E cells in vitro cultured were randomly divided into 6 groups, i.e., the normal control group (Group C), the TGF-beta1 induced model group (Group T), the low dose CA treated group (Group E1), the moderate dose CA treated group (Group E2), the high dose CA group (Group E3), and the Benazepril Hydrochloride Tablet treated group (Group Y). Except Group C, corresponding medication (with an action of 48 h) was administered to cells in the rest groups after they were induced by TGF-beta1 for 24 h. The morphological changes were observed by inverted phase contrast microscope. The distribution of beta-actin protein was detected by immunohistochemical assay. The mRNA expressions of alpha-smooth muscle actin (alpha-SMA) and E-cadherin (E-cad) were detected by real-time PCR. The concentration of fibronectin (FN) was detected by ELISA.

RESULTSAfter induced by TGF-beta1 for three days, hypertrophy and elongated cells in fusiform-shape occurred,with increased expressions of beta-actin protein in the cytoskeletal structure (P < 0.05), bundle fibrous structure occurred inside cytoplasm with significantly up-regulated intracellular alpha-SMA mRNA expressions (P < 0.05), E-cad mRNA expression decreased (P < 0.05), the FN content in the supernate increased (P < 0.05) in Group T. Compared with Group T, partial cells in Group E1, E2, and E3 showed fibrous changes, accompanied with decreased expression of beta-actin protein and FN concentration (P < 0.05). The expression of alpha-SMA mRNA increased and the expression E-cad mRNA decreased in Group E2 and E3 (both P < 0.05). But there was no statistical difference in the expression levels of E-cad and alpha-SMA mRNA (P > 0.05). Compared with Group E1, the expression of beta-actin protein and FN concentration decreased in Group E2 and E3 (P < 0.05). The expressions of alpha-SMA mRNA decreased and E-cad mRNA increased in Group E3 (P < 0.05). Compared with Group Y, the expression of beta-actin mRNA and FN concentration increased in Group E1 (P < 0.05); the expression of beta-actin mRNA increased in Group E3 (P < 0.05); the expression of E-cad mRNA decreased in Group E3 (P < 0.05). There was no statistical difference in the expression of alpha-SMA mRNA among Group E1, E2, and E3 (P > 0.05).

CONCLUSIONCA could inhibit the occurrence of TGF-beta1 induced EMT, which could be used as an effective drug for treating chronic renal insufficiency.

Animals ; Cell Transdifferentiation ; drug effects ; Cells, Cultured ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; Kidney Tubules ; cytology ; Male ; Myofibroblasts ; drug effects ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effect of small interfering RNA (siRNA)-mediated islet neogenesis associated protein (INGAP) gene silencing on the proliferation of islet cells.

METHODSDifferent siRNAs targeting INGAP gene were designed and transfected into INS-1 islet cells, and the expression levels of INGAP mRNA and protein following the transfection were detected using RT-PCR, flow cytometry and Western blotting. The proliferation of the transfected INS-1 cells was evaluated using MTT assay.

RESULTSCompared with those in the irrelevant siRNA, empty vector control, and un-transfected groups, the expression levels of INGAP mRNA and protein in the cells transfected with siRNA6 were reduced significantly. The cell proliferation rate significantly increased after transfection with siRNA6 (P<0.05).

CONCLUSIONsiRNA targeting INGAP can effectively down-regulate INGAP expression and inhibit the proliferation of INS-1 cells.

Animals ; Antigens, Neoplasm ; genetics ; Biomarkers, Tumor ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Insulinoma ; pathology ; Islets of Langerhans ; pathology ; Lectins, C-Type ; genetics ; Pancreatitis-Associated Proteins ; RNA Interference ; RNA, Small Interfering ; genetics ; Rats

OBJECTIVETo investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent.

METHODSHuman DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test.

RESULTSAn exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection.

CONCLUSIONMicrobubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.

Adaptor Proteins, Signal Transducing ; genetics ; Cells, Cultured ; Contrast Media ; administration & dosage ; pharmacology ; Cytoskeletal Proteins ; genetics ; Dendritic Cells ; drug effects ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; Microbubbles ; Plasmids ; Transfection ; Ultrasonics

Objective To explore the isometry of grafts in PCL(posterior cruciate ligament)double-bundle re-construction under femoral tunnel shifting condition.Method Knee specimens from ten fresh frozen cadavers were used.PCL were divided into anterolateral bundles(ALB)and posteromedial bundles(PMB)to the inser-tion footorint.The anterior,postedor,proximal,distal and central points of the two bundles'femoral attachment site were respectivelyanchored to the middle of the PCL's tibial attachment site by the trial wires.Changes in length of the intra-articular part of the wires were recorded while the knee was flexed from 0°to 120°.Result The length changes in every point were compared.All of the maximal length changes of ALB's proximal,pos-todor points and PMB's proximal points were not greater than 2mm.No significant difference between the length changes of ALB's proximal point and posterior(P=0.864>0.05)was found.Conclusions The femo-ral tunnel for the PCL double-bundle reconstruction should be located as follows:ALB should be at the middle point of upper edge of femoral attachment site(proximal point),while PIVIB at the middle point of femoral attachment site(proximal point).

Objective To explore the isometry of grafts in PCL(posterior cruciate ligament)double-bundle re-construction under femoral tunnel shifting condition.Method Knee specimens from ten fresh frozen cadavers were used.PCL were divided into anterolateral bundles(ALB)and posteromedial bundles(PMB)to the inser-tion footorint.The anterior,postedor,proximal,distal and central points of the two bundles'femoral attachment site were respectivelyanchored to the middle of the PCL's tibial attachment site by the trial wires.Changes in length of the intra-articular part of the wires were recorded while the knee was flexed from 0°to 120°.Result The length changes in every point were compared.All of the maximal length changes of ALB's proximal,pos-todor points and PMB's proximal points were not greater than 2mm.No significant difference between the length changes of ALB's proximal point and posterior(P=0.864>0.05)was found.Conclusions The femo-ral tunnel for the PCL double-bundle reconstruction should be located as follows:ALB should be at the middle point of upper edge of femoral attachment site(proximal point),while PIVIB at the middle point of femoral attachment site(proximal point).

BACKGROUNDDiagnosis and treatment for respiratory symptoms (RSs) of gastroesophageal reflux disease (GERD) is more difficult than that for common esophageal symptoms. The goal of this study was to evaluate the efficacy and safety of radiofrequency (RF) treatment on RSs of GERD in a preliminary 12-month follow-up observation.

METHODSFrom April 2006 to October 2008, 505 GERD patients with mainly respiratory presentations such as wheezing, chronic cough or hoarseness, were treated by endoscopic RF. A questionnaire was completed before and after treatment, using a six-point scale ranging from 0 to 5 to assess symptom severity and frequency. The symptom score was the sum of frequency and severity.

RESULTSSymptom scores were significantly improved at the end of the follow-up period. The mean heartburn score decreased from 5.31 to 1.79. The mean regurgitation score decreased from 5.02 to 1.64; mean cough score decreased from 6.77 to 2.85; mean wheezing score decreased from 7.83 to 3.07; and mean hoarseness score decreased from 5.13 to 1.81 (P < 0.01). No major complications or deaths occurred. Minor complications included temporary post-procedural retrosternal unease or pain (n = 106; 21.0%), mild fever (n = 86; 17.0%), transient nausea/vomiting (n = 97; 19.2%), and transient dysphagia (n = 42; 9.3%). Thirty-five (6.9%) patients had recurrence of symptoms. Endoscopic RF treatment was repeated in six patients, and laparoscopic fundoplication was performed in seven.

CONCLUSIONEndoscopic RF is an effective and safe means to treat RSs in patients with GERD.

Adult ; Aged ; Cough ; surgery ; Esophagogastric Junction ; physiopathology ; radiation effects ; Esophagoscopy ; methods ; Female ; Gastroesophageal Reflux ; physiopathology ; surgery ; Heartburn ; surgery ; Hoarseness ; surgery ; Humans ; Male ; Middle Aged ; Radio Waves ; Treatment Outcome ; Young Adult

Objective To investigate the effects of 5-hydroxytryptamine-2C subunit receptor (5-HT 2C R)on long-term potentiation (LTP)of V1M region visual cortex of form deprivation adult amblyopia rats. Methods Sixteen two weeks old Sp-argue Dawley rats were randomly divided into normal control group and monocular form deprivation group,with 8 rats in each group. The rats in the normal control group were not given any intervention;the rats in the monocular form deprivation group were sutured the right eye lid to establish the monocular form deprivation amblyopia model. All rats were fed for 6 weeks after establishing the model successfully,then the rats in the two groups were sacrificed and the coronal examination of 400 μm thick cortical brain slices were incubated in artificial cerebrospinal fluid artificial cerebrospinal fluid (ACSF). According to the difference of drugs in ACSF,the visual cortex slices of rats in normal control group were selected as group A;the contralateral visual cortex slices of the deprivation eye were divided into group B,group C,group D and group E;the ipsilateral visual cortex slices of the deprivation eye were divided into group F,group G,group H and group I. The ACSF of group A,B and F did not added any drugs;the ACSF of group C and group G were added with physiological saline;the ACSF of group D and group H were added with 10 μmol · L - 15-hydroxytryptamine hydrochloride;the ACSF of group E and group I were added with 10 μmol·L - 1 SB 242084 and 10 μmol·L - 15-hydroxytryptamine hydrochloride. The electrophysiology experiment was per-formed in all of the visual cortex slices by extracellular microelectrode recording and the visual cortex fidd postsynaptic poten-tial(fPSP)slope of V1M region of the visual cortex was recorded. Results The fPSP in group A,B,C,D,E,F,G,H,I was (198. 1 ± 13. 5)％,(106. 3 ± 8. 3)％,(106. 3 ± 8. 3)％,(157. 1 ± 9. 7)％,(102. 6 ± 4. 7)％,(144. 5 ± 2. 9)％,(144. 5 ± 2. 9)％,(192. 2 ± 8. 6)％ and (129. 7 ± 13. 5)％,respectively. There was statistic difference in fPSP slope of visual cortex a-mong the group A,B,F(P < 0. 001);the fPSP slope of visual cortex of rats in group A was significantly higher than that in the group B and group F(P < 0. 001);the fPSP slope of visual cortex of rats in group B was significantly lower than that in the group F(P < 0. 001). The fPSP slope of visual cortex in group D was significantly higher than that in the group C (t = - 10. 833,P < 0. 001);the fPSP slope of visual cortex in group H was significantly higher than that in the group D and group G(t = - 6. 841,- 10. 616;P < 0. 001). The fPSP slope of visual cortex in group E was significantly lower than that in the group D and group I(t = 11. 872,- 3. 910;P < 0. 001,P < 0. 05);the fPSP slope of visual cortex in group I was signifi-cantly lower than that in the group H(t = 9. 911,P < 0. 001). Conclusion Monocular deprivation can lead to the dysfunction of bilateral visual cortex neurons and 5-hydroxytryptamine hydrochloride can reverse this phenomenon through 5-HT2C R.

BACKGROUND AND OBJECTIVEStudies have shown that nitric oxide (NO) derived from endothelial nitric oxide synthase (eNOS) is expressed widely in tumor tissues and regulates tumor angiogenesis. However, the results are controversial. This study was to investigate the effect of NO on tumor angiogenesis and its mechanism.

METHODSC57BL/6 mice inoculated with Lewis lung cancer cells were randomly divided into three groups. Mice in the NO group were inoculated with lung cancer cells transfected with eNOS gene, mice in the L-NAME group with L-NAME, an eNOS antagonist, and mice in the control group with normal saline. Plasma concentration of NO and the number of endothelial progenitor cells (EPCs) in peripheral blood were detected . Tumor vessel density, CD133+ cells, and the expression of VEGF-VEGFR in tumor tissues were also measured.

RESULTSFour weeks after inoculation of Lewis cells, tumor volume was significantly larger in control group [ (3022 +/- 401) mm(3)] than in the L-NAME group [ (1204 +/-97) ) mm(3)] and in the eNOS group [(1824 +/- 239) mm(3)] (P<0.01). eNOS protein and NO production increased significantly in Lewis lung cancer cells transfected with eNOS gene. But the number of CD133-positive cells and vessel density in tumors were significantly lower in the eNOS group than in the control group [(48+/-19) / HPF vs. ( 103 +/- 27)/ HPF, (19+/- 7) HPF vs. (31 +/- 9) HPF, P<0.05]. The number of EPCs in peripheral blood was not statistically different between each group. The levels of NO in blood and tumor tissue significantly decreased after the treatment of L-NAME, while the tumor vessel density reduced to 12+/- 5/ HPF (P<0.01, vs. the control group; P<0.05, vs the eNOS transfected group). The number of EPCs in blood and that of CD133-positive cells in tumor tissue were significantly smaller in the L-NAME group than in the control group (P<0.05).

CONCLUSIONNo derived from eNOS inhibits angiogenesis and tumor growth, which may be due to its suppression on either the mobilization or homing of EPCs via VEGF binding to VEGFR.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Carcinoma, Lewis Lung ; blood supply ; metabolism ; pathology ; Cell Count ; Cells, Cultured ; Endothelium, Vascular ; pathology ; Enzyme Inhibitors ; pharmacology ; Female ; Glycoproteins ; metabolism ; Mice ; Mice, Inbred C57BL ; Microvessels ; pathology ; NG-Nitroarginine Methyl Ester ; pharmacology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Nitric Oxide ; blood ; metabolism ; Nitric Oxide Synthase Type III ; antagonists & inhibitors ; genetics ; metabolism ; Peptides ; metabolism ; Plasmids ; Random Allocation ; Stem Cells ; metabolism ; pathology ; Transfection ; Tumor Burden ; Vascular Endothelial Growth Factor A ; blood ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism