Phylogenomic analyses reveal reticulate evolution between Neomicrocalamus and Temochloa (Poaceae: Bambusoideae).

Neomicrocalamus and Temochloa are closely related to bamboo genera. However, when considered with newly discovered and morphologically similar material from China and Vietnam, the phylogenetic relationship among these three groups was ambiguous in the analyses based on DNA regions. Here, as a means of investigating the relationships among the three bamboo groups and exploring potential sources of genomic conflicts, we present a phylogenomic examination based on the whole plastome, single-nucleotide polymorphism (SNP), and single-copy nuclear (SCN) gene datasets. Three different phylogenetic hypotheses were found. The inconsistency is attributed to the combination of incomplete lineage sorting and introgression. The origin of newly discovered bamboos is from introgressive hybridization between Temochloa liliana (which contributed 80.7% of the genome) and Neomicrocalamus prainii (19.3%), indicating that the newly discovered bamboos are closer to T. liliana in genetics. The more similar morphology and closer distribution elevation also imply a closer relationship between Temochloa and newly discovered bamboos.

Khoonmengia honbaensis, a new genus and species of temperate bamboo (Poaceae, Bambusoideae) from central-southern Vietnam.

A new genus of Arundinarieae, Khoonmengia, is established to accommodate a unique new bamboo species, K. honbaensis, from central-southern Vietnam. The morphological features, habitats and distribution of Khoonmengia and related genera, i.e. Ampelocalamus and Hsuehochloa, are compared. The characters of its scrambling habit, internodes with brownish green dots, conspicuous nodes swollen at one side, elliptic buds wholly sunken into culm, extravaginal branching pattern, mid-culm branch complement with one central dominant branch elongating to reiterate the culm accompanied by several lateral slender branches, swollen culm sheath base with a distinctive zone of transverse wrinkles, synflorescence composed of only one spikelet, single or several to many synflorescences arranged into a raceme or panicle terminal on leafy branches, purple anthers and nut-like caryopsis with hardened pericarp and loosely adherent lemma and palea distinguish K. honbaensis from morphologically similar taxa. In order to investigate the phylogenetic position of this unknown bamboo, molecular phylogenetic analyses based on the nuclear gene GBSSI were also conducted, and the results proved that K. honbaensis is definitely a member of Arundinarieae with an isolated position, which also indicated that this species could not be assigned to any of the already described genera and supported the establishment of the new genus.

Chemical constituents from Dendrobium hainanense.

A new phenolic derivative (1) and a new dihydrophenanthrene (2) were isolated from the aerial part of Dendrobium hainanense rofe, along with 12 known compounds. The structures of the new compounds were elucidated by spectroscopic analysis，and the relative configuration of compound 1 was determined by J-based configuration analysis (JBCA) method. Bioassay result indicated that compound 1 exhibited weak antibacterial activity against Canidia albicans and Ralstonia solanaceanum.

Iodine regulates G2/M progression induced by CCL21/CCR7 interaction in primary cultures of papillary thyroid cancer cells with RET/PTC expression.

Treatment with high iodine concentrations can delay oncogenic activation effects, reduce cell growth and return thyroid-specific gene and protein expression levels to normal. During rearranged during transfection (RET)/papillary thyroid carcinoma (PTC) 3 activation, excess iodine can act as a protective agent in thyroid follicular cells. The chemokine receptor CCR7 serves a critical role in lymphocyte trafficking into and within lymph nodes, the preferential metastatic site for PTC. However, the potential associations between chemokine (C­C motif) ligand 21 (CCL21)/C­C chemokine receptor type 7 (CCR7) interaction and iodine concentrations in primary cultures of PTC with RET/PTC expression remain unclear. Proliferation assays of primary cultures of PTC cells with RET/PTC1 and RET/PTC3 expression indicated that CCR7 activation by its specific ligand, CCL21, was associated with significantly increased cell proliferation. Flow cytometry data indicated that CCL21/CCR7 interaction significantly increased the fraction of cells in the G2/M phase of the cell cycle. Western blotting indicated that CCL21/CCR7 interaction significantly upregulated cyclin A, cyclin B1 and cyclin­dependent kinase 1 (CDK1) expression. Western blotting determined that CCL21/CCR7 interaction significantly enhanced the levels of phosphorylated extracellular signal­regulated kinase (P­ERK). Co-immunoprecipitation confirmed that there was interaction between P­ERK and cyclin A, cyclin B1 or CDK1, particularly in the presence of CCL21. Sodium iodide (NaI, 10-5 M) significantly abolished the effects of exogenous CCL21. These results suggest that CCL21/CCR7 interaction contributes to G2/M progression of RET/PTC­expressing cells via the ERK pathway in association with 10­5 M NaI.

The miR-200 family regulates the epithelial-mesenchymal transition induced by EGF/EGFR in anaplastic thyroid cancer cells.

The miR-200 family was recently identified as a suppressor of epithelial-mesenchymal transition (EMT). The loss or gain of miR-200 family members is associated with cancer invasion. The epidermal growth factor receptor (EGFR) is overexpressed in the majority of anaplastic thyroid cancers (ATCs). The activation of EGFR by its ligand, epidermal growth factor (EGF), activates a signaling cascade that results in the enhanced migration and invasiveness of thyroid cancer cells. However, little is known about the potential interrelationships between EGF/EGFR, miR-200s and the induction of EMT or mesenchymal-epithelial transition (MET) processes. This study aimed to investigate the regulatory role of miR-200s in EMT modulation by EGF/EGFR. Using transfection, real-time reverse transcription PCR and western blot analysis, we found that the EGF treatment of Nthy-ori 3-1 thyroid follicular cells resulted in the downregulation of E-cadherin and the upregulation of vimentin. By contrast, EGFR silencing in SW1736 human thyroid carcinoma cells led to the upregulation of E-cadherin and the downregulation of vimentin. In addition, EGF signaling correlated with the reduced expression of miR-200s and the re-expression of miR-200s abrogated the effects of EGF treatment and restored an epithelial phenotype to EGF-induced Nthy-ori 3-1 cells. Conversely, the silencing of miR-200s in SW1736 cells overcame siEGFR-induced changes in gene expression and phenotype. In addition, we demonstrate that miR-200s play a key role in in vitro EGF/EGFR-mediated thyroid cell invasion and in EMT in vivo. We, therefore, provide a mechanistic link between the miR-200 family and EGF/EGFR, which suggests that miR-200 upregulation may serve as a novel therapeutic strategy for highly invasive thyroid cancers.

[Expression and significance of SARI and CCN1 in human colorectal carcinomas].

OBJECTIVE: To explore the expressions of SARI (suppressor of AP-1, regulated by IFN) and connective tissue growth factor/cysteine-rich 61/nephroblastoma-1 (CCN1) and clarify their influences on the occurrence, development and prognosis of colorectal carcinoma (CRC). METHODS: Real-time PCR (polymerase chain reaction) was used to confirm the expressions of SARI and CCN1 at the mRNA level in 32 fresh tissue samples. And the expressions of Caco-2, HT-29 and Lovo were also detected by RT-PCR (reverse transcription-PCR) in cell lines. Tissue specimens were obtained from 116 cases of CRC and the expressions of SARI and CCN1 for each specimen detected by immunohistochemistry. The correlations between the expressions of SARI and CCN1 proteins were summarized. The relationships between the expression levels of SARI and CCN1 and their clinical features in primary CRC were analyzed respectively. The effects of expression levels of SARI and CCN1 proteins on the prognosis were also assessed in 116 CRC cases. RESULTS: The expressions of SARI and CCN1 at the mRNA level in fresh cancerous tissues and cell lines decreased and became up-regulated respectively. The positive rate of SARI protein expression was 76.7% and 28.4% in cancerous and noncancerous tissues respectively (P < 0.05). The positive rate of CCN1 protein expression was 26.7% and 74.1% in cancerous and noncancerous tissues respectively (P < 0.05). A negative correlation was observed between the expressions of SARI and CCN1 (r = -0.24, P < 0.05). The negative expression of SARI correlated with a low grade of differentiation, deep infiltration depth and high TNM staging (P < 0.05). A positive expression of CCN1 correlated with deep infiltration depth and high TNM staging (P < 0.05) while a negative expression of SARI correlated with a lower survival rate than that of a positive expression (χ(2) = 8.47, P < 0.05); additionally, the survival rate of patients with a negative expression of SARI plus a positive expression of CCN1 was further lowered (χ(2) = 12.56, P < 0.05). CONCLUSION: The aberrant expressions of SARI and CCN1 correlate with the malignant biobehaviors of CRC. And a negative expression of SARI correlates with a worse prognosis of CRC.

Telomerase activity and cell apoptosis in colon cancer cell by human telomerase reverse transcriptase gene antisense oligodeoxynucleotide.

AIM: To evaluate the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (As-ODN) on telomerase activity and cell apoptosis in colon cancer cell line SW480. METHODS: As-ODN was transfected into cells SW480 by liposomal transfection. Cultured cells were divided into three groups: ASODN (5'GGAGCGCGCGGCATCGCGGG-3'), sense oligodeoxynucleotide (5'-CCCGCGATGCCGCGCGCTCC-3', S-ODN) and control. The concentration of oligodeoxynucleotide and liposome was 10 micromol/L and 16 mg/L, respectively. The activity of telomerase was examined by telomeric repeat amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA), and cell apoptosis was observed by morphology and flow cytometry in each group. RESULTS: Telomerase activity began to be down-regulated or inhibited when cells SW480 were treated with As-ODN for 72 h, and cell apoptosis was induced. CONCLUSION: It is suggested that hTERT As-ODN might specially inhibit the activity of telomerase in colon cancer cells and it is further proved that the hTERT gene has a significant correlation with telomerase activity. Further evidence is needed to prove whether hTERT As-ODN is a potential tool for the treatment of colon cancer.

Expression and significance of PTEN, hypoxia-inducible factor-1 alpha in colorectal adenoma and adenocarcinoma.

AIM: To investigate the expression and significance of PTEN, hypoxia-inducible factor-1 alpha (HIF-1alpha), and targeting gene VEGF during colorectal carciogenesis. METHODS: Total 71 cases colorectal neoplasms (9 cases of colorectal adenoma and 62 colorectal adenocarcinoma) were formalin fixed and paraffin-embedded, and all specimens were evaluated for PTEN mRNA, HIF-1alpha mRNA and VEGF protein expression. PTEN mRNA, HIF-1alpha mRNA were detected by in situ hybridization. VEGF protein was identified by citrate-microwave SP immunohistochemical method. RESULTS: There were significant differences in PTEN, HIF-1alpha and VEGF expression between colorectal adenomas and colorectal adenocarcinoma (P<0.05). The level of PTEN expression decreased as the pathologic stage increased. Conversely, HIF-1alpha and VEGF expression increased with the Dukes stage as follows: stage A (0.1029+/-0.0457: 0.1207+/-0.0436), stage B (0.1656+/-0.0329: 0.1572+/-0.0514), and stage C+D (0.2335+/-0.0748: 0.2219+/-0.0803). For PTEN expression, there was a significant difference among Dukes stage A, B, and C+D, and the level of PTEN expression was found to be significant higher in Dukes stage A or B than that of Dukes stage C or D. For HIF-1alpha expression, there was a significant difference between Dukes stage A and B, and the level of HIF-1alpha expression was found to be significantly higher in Dukes stage C+D than that of Dukes stage A or B. The VEGF expression had similar results as HIF-1alpha expression. In colorectal adenocarcinoma, decreased levels of PTEN were significantly associated with increased expression of HIF-1alpha mRNA (r=-0.36, P<0.05) and VEGF protein (r=-0.48, P<0.05) respectively. The levels of HIF-1 were positively correlated with VEGF expression (r=0.71, P<0.01). CONCLUSION: Loss of PTEN expression and increased levels of HIF-1alpha and VEGF may play an important role in carcinogenesis and progression of colorectal adenocarcinoma.

Mast cell density and the context of clinicopathological parameters and expression of p185, estrogen receptor, and proliferating cell nuclear antigen in gastric carcinoma.

AIM: To investigate the relationship between the mast cell density (MCD) and the context of clinicopathological parameters and expression of p185, estrogen receptor (ER), and proliferating cell nuclear antigen (PCNA) in gastric carcinoma. METHODS: Mast cell, p185, ER, and PCNA were detected using immunohistochemical S-P labeling method. Mast cell was counted in tissue of gastric carcinoma and regional lymph nodes respectively, and involved lymph nodes (ILN) were examined as usual. RESULTS: MCD was significantly related to both age and depth of penetration (chi(2)=4.688,P<0.05 for age and chi(2)=9.350, P<0.01 for depth of penetration) between MCD>21/0.03 mm(2) and MCD< or =21/0.03 mm(2) in 100 patients; MCD in 1-6 ILN group patients was significantly higher than that in 7-15 ILN or >15 ILN group patients (u=6.881, 8.055, P<0.01); There were significant differences intergroup in positive expression rate of p185, ER and PCNA between MCD>21/0.03 mm(2) and MCD< or =21/0.03 mm(2) in 100 patients. CONCLUSION: Mast cell may have effect on inhibiting invasive growth of tumor, especially in the aged patients; The number of mast cells, in certain degree, may predicate the number of involved lymph nodes, which is valuable for assessment of prognosis; MCD was related to the expression of p185, ER, and PCNA in gastric carcinoma. It suggests that mast cell accumulation may inhibit the proliferation and the dissemination of the gastric carcinoma.