[Knockdown of ACC1 promotes migration of esophageal cancer cell].

Objective: To investigate the effect of acetyl-CoA carboxylase 1 (ACC1) knockdown on the migration of esophageal squamous cell carcinoma (ESCC) KYSE-450 cell and underlying mechanism. Methods: Lentiviral transfection was conducted to establish sh-NC control cell and ACC1 knocking down cell (sh-ACC1). Human siRNA HSP27 and control were transfected by Lipo2000 to get si-HSP27 and si-NC. The selective acetyltransferase P300/CBP inhibitor C646 was used to inhibit histone acetylation and DMSO was used as vehicle control. Transwell assay was performed to detect cell migration. The expression of HSP27 mRNA was examined by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and the expressions of ACC1, H3K9ac, HSP27 and epithelial-mesenchymal transition-related proteins E-cadherin and Vimentin were detected by western blot. Results: The expression level of ACC1 in sh-NC group was higher than that in sh-ACC1 group (P<0.01). The number of cell migration in sh-NC group was (159.00±24.38), lower than (361.80±26.81) in sh-ACC1 group (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC group were statistically significant compared with sh-AAC1 group (P<0.05). The migrated cell number in sh-NC+ si-NC group was (189.20±16.02), lower than (371.60±38.40) in sh-ACC1+ si-NC group (P<0.01). The migrated cell number in sh-NC+ si-NC group was higher than that in sh-NC+ si-HSP27 group (152.40±24.30, P<0.01), and the migrated cell number in sh-ACC1+ si-NC group was higher than that in sh-ACC1+ si-HSP27 group (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC+ si-NC group were significantly different from those in sh-ACC1+ si-NC and sh-NC+ si-HSP27 groups (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-ACC1+ si-NC group were significantly different from those in sh-ACC1+ si-HSP27 group (P<0.01). After 24 h treatment with C646 at 20 µmmo/L, the migrated cell number in sh-NC+ DMSO group was (190.80±11.95), lower than (395.80±17.10) in sh-ACC1+ DMSO group (P<0.01). The migrated cell number in sh-NC+ DMSO group was lower than that in sh-NC+ C646 group (256.20±23.32, P<0.01). The migrated cell number in sh-ACC1+ DMSO group was higher than that in sh-ACC1+ C646 group (87.80±11.23, P<0.01). The protein expressions of H3K9ac, HSP27, E-cadherin and Vimentin in sh-NC+ DMSO group were significantly different from those in sh-ACC1+ DMSO group and sh-NC+ C646 group (P<0.01). The protein expression levels of H3K9ac, HSP27, E-cadherin and Vimentin in sh-ACC1+ DMSO group were significantly different from those in sh-ACC1+ C646 group (P<0.01). Conclusion: Knockdown of ACC1 promotes the migration of KYSE-450 cell by up-regulating HSP27 and increasing histone acetylation.

Body height and the spread of spinal anaesthesia for caesarean section: a prospective controlled trial.

BACKGROUND: No conclusive evidence exists on the effect of patient height on the spread of spinal anaesthesia. Our aim was to measure the ED50 and ED95 values of intrathecal ropivacaine in taller and shorter patients, and thus investigate the hypothesis that the spinal dose requirement in shorter patients is lower than that in taller patients undergoing caesarean section. METHODS: In this study, 270 pregnant women were assigned to the taller (Group T) or shorter group (Group S) based on their heights. Subjects in both groups were further randomly assigned to one of nine subgroups based on the dosage of intrathecal isobaric ropivacaine to be administered (7, 8, 9, 10, 11, 12, 13, 14 or 15 mg respectively). RESULTS: The ED50 and ED95 values of ropivacaine were 9.24 mg and 13.36 mg in Group S, and 10.11 mg and 14.63 mg in Group T, with no inter-group difference (P = 0.886). There was a significant inter-group difference in the incidence of hypotension and the changes in mean arterial pressure after spinal anaesthesia using 15 mg ropivacaine. The dose of ephedrine administered in Group S was higher than that in Group T when 15 mg ropivacaine was administered (P = 0.031). CONCLUSION: The taller and shorter patients did not respond differently to modest intrathecal doses of ropivacaine. However, a larger dose of ropivacaine was associated with an increased incidence of hypotension in shorter patients compared to that in taller patients.

Molecular cloning and expression pattern of the DjStag gene in the planarian Dugesia japonica during embryonic development.

We examined STAG-related gene (DjStag) expression in the planarian Dugesia japonica. This species is common in Far Eastern countries. The DjStag cDNA includes 1362 bp and contains a 489-bp open reading frame corresponding to a deduced protein of 162 amino acids, with a 170-bp 5'-UTR and a 703-bp 3'-UTR. Phylogenetic analysis showed that DjStag is an STAG/STAG-like member. We examined the expression pattern of DjStag in this planarian during embryonic development by whole-mount in situ hybridization. DjStag was detected in embryonic cells in the germ band at early embryo stages. The number of DjStag-positive embryonic cells increased in stage 5. Later, it was mainly expressed in lateral region parenchyma. In juveniles, extensive expression of DjStag was observed not only in the head and tail regions, but also in the parenchyma between the epidermis and the gastrodermis. We conclude that DjStag is expressed in the cellular subset that will become the neoblast cells of the adult flatworm. DjStag may play an essential role in spatial and temporal regulation during planarian embryonic development.

Implications of microRNA-197 downregulated expression in esophageal cancer with poor prognosis.

The aim of this study was to investigate the significance of the microRNA miR-197 expression level in relation to clinicopathological factors and prognoses of esophageal cancer (EC). MicroRNA was extracted using the Taqman(®) MicroRNA Assay from 46 EC patients at the same tumor node metastasis (TNM) stage, but with different prognoses, who underwent surgery. Paracancerous normal tissues were used as controls. The correlation between miR-197 expression and clinicopathologic features was analyzed, and the significance of miR-197 as a prognostic factor and its relationship with survival was determined. miR-197 expression was lower in patients with poor prognosis than in those with good prognosis (P < 0.05). Kaplan-Meier analysis results showed that the miR-197 expression level is significantly correlated with survival time (P = 0.030), and that patients with higher expression of miR-197 had longer survival times. Cox multi-factor model analysis showed that patient prognosis (P = 0.001), tumor length (P = 0.010) and expression (P = 0.042), and survival time were significantly correlated, with corresponding risks of 9.183, 2.318, and 1.925, respectively. This study supports a role of miR-197 as an anti-oncogene and a biomarker for EC and its relationship with other prognostic factors and survival.

Overregulation of microRNA-212 in the poor prognosis of esophageal cancer patients.

There have been few reports evaluating the expression and function of the microRNA miR-212 in esophageal cancer. The aim of this study was to investigate the relationship between miR-212 expression and clinicopathological factors and prognoses of esophageal cancer. MicroRNA was extracted from 46 esophageal cancer patients using the Taqman MicroRNA assay. All patients were at the same tumor node metastasis stage, but with different prognoses, and had all undergone surgery. The correlation between miR-212 expression and clinicopathological features was analyzed and the significance of miR-212 as a prognostic factor as well as its relationship with survival was determined. miR-212 expression was higher in patients with poor prognoses than in those with good prognoses (P < 0.0001). Kaplan-Meier analysis results showed that the miR-212 expression level was significantly correlated with survival time (P = 0.024). Patients with higher expression of miR-212 showed longer survival times. Cox multi-factor model analysis showed that miR-212 expression was significantly correlated with survival time (P = 0.026). mir-212 is related with prognostic factors and survival time and may be a biomarker for esophageal cancer.

Expression patterns of the STAG gene in intact and regenerating planarians (Dugesia japonica).

We examined the spatial and temporal expression of the planarian Dugesia japonica STAG-related gene (DjStag), in both intact and regenerating planarians, by whole-mount in situ hybridization and relative quantitative real-time PCR. The first localized transcripts of DjStag were detected in the blastemas three days after amputation, in all regenerates including those from head, tail and trunk pieces. The maximum level of expression of DjStag transcripts occurred at five days after cutting. After regeneration for seven days, DjStag was weakly expressed. A similar decrease occurs regardless of the orientation of the cut. The expression pattern did not differ significantly in the different types of regeneration. Relative quantitative real-time PCR analysis of DjStag mRNA indicated that the expression of DjStag mRNA was increased after amputation compared to that in normal intact planarians, and the maximum level of expression of DjStag transcripts occurred at five days after amputation. All results suggest that DjStag, implicated in planarian regeneration, plays a role in maintaining the ability of pluripotent stem cells to regenerate lost tissue in planarians.

Expression pattern of DjPreb gene during the planarian Dugesia japonica embryonic development.

Prolactin regulatory element binding (PREB) protein belongs to the family of WD-repeat proteins which are regulatory and versatile proteins for diverse functions. In this study we have shown the expression pattern of the planarian Dugesia japonica PREB-related gene (DjPreb) during embryonic development by whole-mount in situ hybridization. Genomic analysis reveals that the DjPreb gene consists of two exons and one intron. Expression of the DjPreb mRNA was not observed as early as in stage 3 embryos. DjPreb positive signal was first found in stage 4. It is expressed in some embryonic cells in the periphery of the embryo. The number of DjPreb positive embryonic cells grows in stage 5. DjPreb is expressed in the dorsolateral regions and part of the anterior regions in stage 6. In stage 7, DjPreb positive signals are detected in the dorsolateral regions along the A-P axis and from stage 8 to the juvenile stage DjPreb mRNA is strongly expressed not only in the differentiating tissues of the anterior and posterior regions, but also in the parenchyma of the dorsolateral regions, and generates the gradient in the head of the juvenile. These results on the DjPreb expression pattern suggest its potential role in the specification of many cell types; in particular, DjPreb may play an essential role in spatial and temporal regulation during the head and tail formation and the anterior/posterior patterning formation.

[Characterization and expression of DjPreb gene in the planarian Dugesia japonica].

In this study we report the expression and identification of a PREB-related gene from the planarian Dugesia japonica, DjPreb. The planarian DjPreb cDNA is comprised of 1101 bp and contains a 972 bp open reading frame corresponding to a deduced protein of 323 amino acids with a 69 bp 5'-UTR and a 60 bp 3'-UTR. Phylogenetic analysis shows that DjPreb belongs to PREB/PREB-like members. We examined its spatial and temporal expression and distribution in both intact and regenerating planarians by Relative quantitative real-time PCR and Whole-mount in situ hybridization. The analysis indicates that DjPreb shows a gradient of expression with peak levels present in the anterior and posterior regions and progressively lower levels in central regions in intact and regenerating planarians. During regeneration the expression of DjPreb is upregulated. Strong expression of DjPreb is observed in the anterior and posterior blastemas. These results suggest that DjPreb may participate in head and tail formation.