Cathepsin L induced PC-12 cell apoptosis via activation of B-Myb and regulation of cell cycle proteins.

Cathepsin L (CTSL), a cysteine protease, is responsible for the degradation of a variety of proteins. It is known to participate in neuronal apoptosis associated with abnormal cell cycle. However, the mechanisms underlying CTSL-induced cell apoptosis remain largely unclear. We reported here that rotenone caused an activation of CTSL expression in PC-12 cells, while knockdown of CTSL by small interfering RNAs or its inhibitor reduced the rotenone-induced cell cycle arrest and apoptosis. Moreover, elevation of CTSL and increased-apoptosis were accompanied by induction of B-Myb, a crucial cell cycle regulator. We found that B-Myb was increased in rotenone-treated PC-12 cells and knockdown of B-Myb ameliorated rotenone-stimulated cell apoptosis. Further analysis demonstrated that CTSL influenced the expression of B-Myb as suppression of CTSL activity led to a decreased B-Myb expression, whereas overexpression of CTSL resulted in B-Myb induction. Reduction of B-Myb in CTSL-overexpressing cells revealed that regulation of cell cycle-related proteins, including cyclin A and cyclin B1, through CTSL was mediated by the transcription factor B-Myb. In addition, we demonstrated that the B-Myb target, Bim, and its regulator, Egr-1, which was also associated with CTSL closely, were both involved in rotenone-induced apoptosis in PC-12 cells. Our data not only revealed the role of CTSL in rotenone-induced neuronal apoptosis, but also indicated the involvement of B-Myb in CTSL-related cell cycle regulation.

(E)-4-Amino-N'-(3-nitro-benzyl-idene)benzohydrazide.

In the title compound, C(14)H(12)N(4)O(3), the dihedral angle between the benzene rings is 7.6 (4)°. In the crystal, infinite sheets linked by N-H⋯O and bifurcated N-H⋯(O,N) hydrogen bonds propagate in the (10-1) plane, in which R(4) (4)(36) loops are apparent. Neighbouring layers may inter-act by way of very weak π-π stacking inter-actions [centroid-centroid distances = 3.9329 (13) and 4.0702 (13) Å].

[Optimization of extraction on flavonoids from leaves of Heritiera littoralis].

OBJECTIVE: To optimize the preparation procedure for extraction of flavonoids from leaves of Heritiera littoralis. METHODS: Comparing microwave extraction and ultrasonic extraction, The content of total flavonoids was measure by UV spectrophometry and the wavelength set at 510 nm. The effects of treatment time, extraction power, ethanol concentration and solid-liquid ratio were studied. The orthogonal test of four factors and three levels were studied to obtain the extraction technology of flavonoids from leaves of Heritiera littoralis. RESULTS: The optimal technical conditions were mixing leaves of Heritiera littoralis with 70% ethanol on the ratio of 1 g: 30 mL, when microwave treatment time was 8 min and micro-wave power was 400 W. Under this condition, the extraction rate reached 6.15%. CONCLUSION: This study explores a practical extraction method for flavonoids from leaves of Heritiera littoralis, and lay a good foundation for mangrove medical research in future.

Treatment of nontraumatic osteonecrosis of the femoral head with the implantation of core decompression and concentrated autologous bone marrow containing mononuclear cells.

BACKGROUND: Since self-limited repair ability of the necrotic lesion may be a cause for failure of the technique, the possibility has been raised that bone marrow mononuclear cells (BMMCs) containing BMSCs implanted into a necrotic lesion of the femoral head with core decompression (CD) may be of benefit in the treatment of this condition. For this reason, we studied the implantation of the concentrated autologous bone marrow containing mononuclear cells in necrotic lesion of the femoral head to determine the effect of the method. METHODS: The study included 45 patients (59 hips, 9 females, 36 males; mean age 37.5 years, range 16-56 years) with stages I-IIIA nontraumatic avascular necrosis of the femoral head according to the system of the Association Research Circulation Osseous. Concentrated bone marrow (30-50 ml) containing mononuclear cells has been gained from autologous bone marrow (100-180 ml) obtained from the iliac crest of patient with the cell processor system. Concentrated bone marrow was injected through a CD channel into the femoral head. The outcome was determined by the changes in the Harris hip score, by progression in radiographic stages, and by the need for hip replacement. The mean follow-up was 27.6 months (range 12-40 months). RESULTS: Pre- and post-operative evaluations showed that the mean Harris hip score increased from 71 to 83. Clinically, the overall success is 79.7%, and hip replacement was done in 7 of the 59 hips (11.9%). Radiologically, 14 of the 59 hips exhibited femoral head collapse or narrowing of the coxofemoral joint space, and the overall failure rate is 23.7%. The number of BMMCs increased from 12.2 +/- 3.2 x 10(6)/ml to 35.2 +/- 12 x 10(6)/ml between pre-concentration and post-concentration. CONCLUSION: The concentrated autologous bone marrow containing mononuclear cells implantation relieves hip pain, prevents the progression of osteonecrosis. Therefore, it may be the treatment of choice particularly in stages I-II nontraumatic osteonecrosis of the femoral head.

Author Correction: Draft genomic and transcriptome resources for marine chelicerate Tachypleus tridentatus.

In the original version of this Data Descriptor the word "Gulf" was incorrectly spelled in the affiliation "Ocean College, Beibu Gulf University, Qinzhou, 535011, Guangxi, China". This has now been corrected in both the HTML and PDF versions.

Draft genomic and transcriptome resources for marine chelicerate Tachypleus tridentatus.

Chinese horseshoe crabs (Tachypleus tridentatus), ancient marine arthropods dating back to the mid-Palaeozoic Era, have provided valuable resources for the detection of bacterial or fungal contamination. However, excessive exploitation for the amoebocyte lysate of Tachypleus has dramatically decreased the population of the Chinese horseshoe crabs. Thus, we present sequencing, assembly and annotation of T. tridentatus, with the hope of understanding the genomic feature of the living fossil and assisting scientists with the protection of this endangered species. The final genome contained a total size of 1.943 Gb, covering 90.23% of the estimated genome size. The transcriptome of three larval stages was constructed to investigate the candidate gene involved in the larval development and validate annotation. The completeness of the genome and gene models was estimated by BUSCO, reaching 96.2% and 95.4%, respectively. The synonymous substitution distribution of paralogues revealed that T. tridentatus had undergone two rounds of whole-genome duplication. All genomic and transcriptome data have been deposited in public databases, ready to be used by researchers working on horseshoe crabs.

5-(1H-1,2,3-Benzotriazol-1-ylmeth-yl)-3-phenyl-1,2,4-oxadiazole.

In the title mol-ecule, C(15)H(11)N(5)O, the 1,2,3-benzotriazole and 3-phenyl-1,2,4-oxadiazole units are individually essentially planar and the dihedral angle between them is 80.2 (2)°. In the crystal structure, mol-ecules are connected via weak inter-molecular C-H⋯N hydrogen bonds to form two-dimensional sheets.

Benzamide oxime.

In the crystal structure of the title compound, C(7)H(8)N(2)O, mol-ecules are connected via inter-molecular N-H⋯O and O-H⋯N hydrogen bonds to form a two-dimensional supra-molecular structure. The oxime group has an E configuration and the dihedral angle between the mean planes of the benzene ring and the amidoxime grouping is 20.2 (3)°.

[Complete sequence and gene organization of the Tibetan chicken mitochondrial genome].

Using PCR amplification, sequencing and assembling, we obtained the complete mitochondrial genome of Tibetan chicken. The complete mitochondrial genome was 16 783 bp in length. It contained 37 genes (13 protein coding genes, 2 rRNA, 22 tRNA) and a control region. The deduced restriction map revealed a unique pattern of Dra I restriction in Tibetan chicken. Phylogenetic trees based on the D-loop locus and the 13 protein coding genes by Neighbor-joining and Maximum Parsimony analysis indicated that the red junglefowl was the direct ancestor of Tibetan chicken and Tibetan chicken was closest to white leghorn and white plymouth rock, although the evolution of Tibetan chicken appeared to be relatively independent from them. A possible explanation is that the ancestor of Tibetan chicken lived in a relatively isolated environment after entering into the high altitude area and developed unique genetic characters.

A mitochondrial genome sequence of the Tibetan antelope (Pantholops hodgsonii).

To investigate genetic mechanisms of high altitude adaptations of native mammals on the Tibetan Plateau, we compared mitochondrial sequences of the endangered Pantholops hodgsonii with its lowland distant relatives Ovis aries and Capra hircus, as well as other mammals. The complete mitochondrial genome of P. hodgsonii (16,498 bp) revealed a similar gene order as of other mammals. Because of tandem duplications, the control region of P. hodgsonii mitochondrial genome is shorter than those of O. aries and C. hircus, but longer than those of Bos species. Phylogenetic analysis based on alignments of the entire cytochrome b genes suggested that P. hodgsonii is more closely related to O. aries and C. hircus, rather than to species of the Antilopinae subfamily. The estimated divergence time between P. hodgsonii and O. aries is about 2.25 million years ago. Further analysis on natural selection indicated that the COXI (cytochrome c oxidase subunit I) gene was under positive selection in P. hodgsonii and Bos grunniens. Considering the same climates and environments shared by these two mammalian species, we proposed that the mitochondrial COXI gene is probably relevant for these native mammals to adapt the high altitude environment unique to the Tibetan Plateau.

Autophagy involvement in olanzapine-mediated cytotoxic effects in human glioma cells.

The aim of this study was to investigate the effects of olanzapine on growth inhibition as well as autophagy in glioma cells in vitro and in vivo. The proliferation of both LN229 and T98 glioma cells, measured by MTT assay, was suppressed in a concentration-dependent and time-dependent manner. Moreover, apoptosis of both cells was significantly increased with the treatment of olanzapine as evidenced by increased Bcl-2 expression, Hoechst 33258 staining and annexinV-FITC/PI staining. Olanzapine treatment also enhanced activation of autophagy with increased expression of LC3-II, expression of protein p62, a substrate of autophagy, being decreased. The growth inhibition by olanzapine in both glioma cell lines could be blocked by co-treatment with 3-MA, an autophagy inhibitor. Furthermore, olanzapine effectively blocked the growth of subcutaneous xenografts of LN229 glioma cells in vivo. The increased level of protein LC3-II and decreased level of p62 followed by a decreased level of Bcl-2, suggesting that autophagy may contribute to apoptosis. In addition, reduced proliferation of glioma cells was shown by a decrease of Ki-67 staining and increased caspase-3 staining indicative of apoptosis in mouse xenografts. These results indicated that olanzapine inhibited the growth of glioma cells accompanied by induction of autophagy and apoptosis both in vitro and in vivo. Olanzapine-induced autophagy plays a tumor-suppressing role in glioma cells.

[Observation on therapeutic effect of auricular point sticking combined with Tongshiji treatment on child ametropic amblyopia].

OBJECTIVE: To compare therapeutic effects of auricular point sticking combined with Tongshiji treatment and simple Tongshiji treatment on child ametropic amblyopia. METHODS: Three hundred children of ametropic amblyopia were classified as mild, moderate and severe groups, 100 cases in each group. The each group was again randomly divided into a simple Tongshiji treatment (control group) and a auricular point sticking combined with Tongshiji treatment group (observation group). Their therapeutic effects were compared. RESULTS: The total effective rate in the observation group were 100.0%, 79.8% and 71.0%, and in the control group were 100.0%, 54.3% and 48.2% respectivety for mild, moderate and severe groups. For the mild amblyopia children, there was no significant difference between the two groups in the therapeutic effect; for the moderate and severe ametropic amblyopia children, the therapeutic effect in the observation group was better than that in the control group (P<0.05). CONCLUSION: Auricular point sticking combined with Tongshiji treatment for child ametropic amblyopia is of characteristics of convenient manipulation, obvious and rapid therapeutic effect.