Systematic Identification and Analysis of OSC Gene Family of Rosa rugosa Thunb.

The oxidosqualene cyclase family of Rosa rugosa (RrOSC) provides a starting point for the triterpenoid pathway, which contributes to the medicinal value of the extraction of tissues of Rosa rugosa. However, the structure and function of key RrOSCs of active triterpenoids remain ambiguous. In this study, a total of 18 RrOSC members with conservative gene structures and motifs were identified based on the genome of Rosa rugosa. The RrOSCs were located on three chromosomes including two gene clusters that derived from gene replication. The phylogenetic relationship divided RrOSCs into six groups, and the RrOSCs of GI and GIV that were represented by lupeol or α-amyrin were identified as likely to include candidate genes for producing active triterpenoids. Considering the high expression or specific-tissue expression of the candidates, RrOSC1, RrOSC10, RrOSC12, and RrOSC18 were considered the key genes. RrOSC12 was identified in vitro as lupeol synthase. The results provided fundamental information and candidate genes for further illustration of the triterpenoid pathway involved in the pharmacological activities of Rosa rugosa.

Transcription Factor RrANT1 of Rosa rugosa Positively Regulates Flower Organ Size in Petunia hybrida.

The flower is the main organ that produces essential oils in many plants. The yield of raw flowers and the number of secretory epidermal cells are the main factors for essential oil production. The cultivated rose species "Pingyin 1" in China was used to study the effect of RrANT1 on floral organ development. Eighteen AP2 transcription factors with dual AP2 domains were identified from Rosa rugosa genome. RrANT1 belonged to euANT. The subcellular localization results showed that RrANT1 protein is localized in the nucleus. The relative expression level of RrANT1 in the receptacle is higher than that in petals in the developmental stages, and both decreased from the initial phase to senescence. Compared with the RrANT1 expression level in petals in the blooming stage, RrANT1 expression level was significant in petals (~48.8) and highest in the receptacle (~102.5) in the large bud stage. It was only highly expressed in the receptacle (~39.4) in the blooming period. RrANT1 overexpression significantly increased petunia flower and leaf sizes (~1.2), as well as flower fresh weight (~30%). The total number of epidermis cells in the petals of overexpressing plants significantly increased (>40%). This study concluded that RrANT1 overexpression can increase the size and weight of flowers by promoting cell proliferation, providing a basis for creating new rose germplasm to increase rose and essential oil yield.

Roles of the SPL gene family and miR156 in the salt stress responses of tamarisk (Tamarix chinensis).

BACKGROUND: Accumulating evidences show that SPLs are crucial regulators of plant abiotic stress tolerance and the highly conserved module miR156/SPL appears to balance plant growth and stress responses. The halophyte Tamarix chinensis is highly resistant to salt tress. SPLs of T. chinensis (TcSPLs) and theirs roles in salt stress responses remain elusive. RESULTS: In this study, we conducted a systematic analysis of the TcSPLs gene family including 12 members belonging to 7 groups. The physicochemical properties and conserved motifs showed divergence among groups and similarity in each group. The microRNA response elements (MREs) are conserved in location and sequence, with the exception of first MRE within TcSPL5. The miR156-targeted SPLs are identified by dual-luciferase reporter assay of MRE-miR156 interaction. The digital expression gene profiles cluster suggested potential different functions of miR156-targeted SPLs vs non-targeted SPLs in response to salt stress. The expression patterns analysis of miR156-targeted SPLs with a reverse expression trend to TcmiR156 suggested 1 h (salt stress time) could be a critical time point of post-transcription regulation in salt stress responses. CONCLUSIONS: Our work demonstrated the post-transcription regulation of miR156-targeted TcSPLs and transcription regulation of non-targeted TcSPLs in salt stress responses, and would be helpful to expound the miR156/SPL-mediated molecular mechanisms underlying T. chinensis salt stress tolerance.

[Preretinal hemorrhage and prognosis following vitrectomy and silicone oil tamponade for severe proliferative diabetic retinopathy].

OBJECTIVE: To examine the prognosis of preretinal hemorrhage following vitrectomy and silicone oil tamponade for severe proliferative diabetic retinopathy. METHODS: Clinical data of 76 cases of proliferative diabetic retinopathy treated with vitrectomy and silicone oil infusion tamponade in Sir Run Run Shaw Hospital from October 2006 to September 2013 were retrospectively reviewed. Intraoperative bleeding,postoperative preretinal bleeding,blood reabsorption time, and preretinal fibrosis were assessed. RESULTS: All preretinal hemorrhage developed within 1 week after surgery, blood was distributed in thin and scattered patterns (32 cases), thick and localized patterns (25 cases) or thick and scattered patterns (19 cases). The preretinal hemorrhage was ceased in 1 day after operation in 35 cases, in 2 days after operation in 18 cases, in two weeks after operation in 23 case. Recurrent hemorrhage occurred within 1 week after operation in 15 cases. Thin blood was largely reabsorbed in about two weeks, and thick blood was largely reabsorbed in about five weeks. Fibrosis tissue was resulted in 15 cases(34.1%) with thick blood. CONCLUSION: Most of preretinal hemorrhage occurs within 1 week after surgery and is reabsorpted with 5 weeks in patients with proliferative diabetic retinopathy undergoing vitrectomy and silicone oil tamponade. The major complication of preretinal bleeding is the formation of preretinal fibrosis.

Molecular cloning and characterization of farnesyl diphosphate synthase from Rosa rugosa Thunb associated with salinity stress.

Rosa rugosa, a renowned ornamental plant, is cultivated for its essential oil containing valuable monoterpenes, sesquiterpenes, and other compounds widely used in the floriculture industry. Farnesyl diphosphate synthase (FPPS) is a key enzyme involved in the biosynthesis of sesquiterpenes and triterpenes for abiotic or biotic stress. In this study, we successfully cloned and characterized a full-length FPPS- encoding cDNA identified as RrFPPS1 using RT-PCR from R. rugosa. Phylogenetic analysis showed that RrFPPS1 belonged to the angiosperm-FPPS clade. Transcriptomic and RT-qPCR analyses revealed that the RrFPPS1 gene had tissue-specific expression patterns. Subcellular localization analysis using Nicotiana benthamiana leaves showed that RrFPPS1 was a cytoplasmic protein. In vitro enzymatic assays combined with GC-MS analysis showed that RrFPPS1 produced farnesyl diphosphate (FPP) using isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) as substrates to provide a precursor for sesquiterpene and triterpene biosynthesis in the plant. Additionally, our research found that RrFPPS1 was upregulated under salt treatment. These substantial findings contribute to an improved understanding of terpene biosynthesis in R. rugosa and open new opportunities for advancements in horticultural practices and fragrance industries by overexpression of the RrFPPS1 gene in vivo increased FPP production and subsequently led to elevated sesquiterpene yields in the future. The knowledge gained from this study can potentially lead to the development of enhanced varieties of R. rugosa with improved aroma, medicinal properties, and resilience to environmental stressors.

Identification of Volatile Compounds and Terpene Synthase (TPS) Genes Reveals ZcTPS02 Involved in ß-Ocimene Biosynthesis in Zephyranthes candida.

Zephyranthes candida is a frequently cultivated ornamental plant containing several secondary metabolites, including alkaloids, flavonoids, and volatile organic compounds (VOCs). However, extensive research has been conducted only on non-VOCs found in the plant, whereas the production of VOCs and the molecular mechanisms underlying the biosynthesis of terpenes remain poorly understood. In this study, 17 volatile compounds were identified from Z. candida flowers using gas chromatography-mass spectrometry (GC-MS), with 16 of them being terpenoids. Transcriptome sequencing resulted in the identification of 17 terpene synthase (TPS) genes; two TPS genes, ZcTPS01 and ZcTPS02, had high expression levels. Biochemical characterization of two enzymes encoded by both genes revealed that ZcTPS02 can catalyze geranyl diphosphate (GPP) into diverse products, among which is ß-ocimene, which is the second most abundant compound found in Z. candida flowers. These results suggest that ZcTPS02 plays a vital role in ß-ocimene biosynthesis, providing valuable insights into terpene biosynthesis pathways in Z. candida. Furthermore, the expression of ZcTPS02 was upregulated after 2 h of methyl jasmonate (MeJA) treatment and downregulated after 4 h of the same treatment.

Integrated Metabolomics and Transcriptomics Analysis Reveals New Insights into Triterpene Biosynthesis in Rosa rugosa.

Rosa rugosa is highly regarded for its aesthetic and therapeutic qualities. In particular, R. rugosa's flowers are known to produce essential oils containing a mixture of volatile terpenes, phenylpropanoids, and other compounds. Despite this, extensive research exists on volatile terpenes in flowers, while the knowledge of non-volatile terpenes in distinct tissues is still limited. Using UPLC-ESI-MS/MS, a comprehensive analysis of the terpene metabolites in five different tissues of R. rugosa was conducted. These metabolites accumulated in distinct tissues, and the majority of them were triterpenoids. Transcriptome data were collected from five tissues using RNA-seq. Transcriptomics and metabolomics were utilized to evaluate the triterpene biosynthesis pathway, resulting in new insights into its regulation and biosynthesis. The RrOSC10 was identified as a key enzyme in converting 2,3-oxidosqualene into α-amyrin, potentially contributing to the triterpene biosynthesis pathway. Furthermore, the expression of the RrOSC10 gene was upregulated by salinity for 0.5 h and 1 h, with subsequent downregulation at 2 h. This study lays a foundation for future research on the biosynthesis and accumulation of triterpenes in R. rugosa.

The evaluation of the retinal nerve fiber layer in multiple sclerosis with special-domain optical coherence tomography.

BACKGROUND/AIMS: Retinal nerve fiber layer (RNFL) thinning has been observed on histopathology and time-domain optical coherence tomography in many diseases of the central nervous system. In this study, with a higher resolution of spectral-domain optical coherence tomography (SDOCT), we detected RNFL changes in patients with multiple sclerosis (MS) in China, and compared RNFL thickness between eyes with and without optic neuritis (ON). METHODS: In this retrospective, nonrandom case study, the patients were recruited from the Affiliated Sir Run Run Shaw Hospital of Zhejiang University. RNFL thickness was measured for each eye using SDOCT. The controls were recruited from the healthy population. RESULTS: Peripapillary RNFL thickness of 24 eyes in 12 patients was detected by SDOCT. The average RNFL thickness of the MS patients was 81.9 ± 17.8 µm compared to the control value of 102.1 ± 8.1 µm (p = 0.00). The average RNFL of the patients with a history of ON was thinner than that of patients without ON (71.8 ± 19.2 µm vs. 92.0 ± 8.5 µm, p = 0.001). CONCLUSION: The RNFL thinning in Chinese patients with MS can be detected by SDOCT. The SDOCT scan represents a high-resolution, objective, noninvasive and easily quantifiable in vivo biomarker of MS.

GT Transcription Factors of Rosa rugosa Thunb. Involved in Salt Stress Response.

Rosa rugosa was a famous aromatic plant while poor salt tolerance of commercial cultivars has hindered its culture in saline-alkali soil. In many plants, the roles of GT (or trihelix) genes in salt stresses responses have been emerging. In the wild R. rugosa, a total of 37 GTs (RrGTs) were grouped into GT-1, GT-2, GTγ, SH4, and SIP1 lineages. SIP1 lineage expanded by transposition. The motifs involved in the binding of GT cis-elements were conserved. Four RrGTs (RrGT11/14/16/18) significantly differentially expressed in roots or leaves under salt stress. The responsive patterns within 8 h NaCl treatment indicated that RrGTγ-4 (RrGT18) and RrGT-1 (RrGT16) were significantly induced by salt in roots of R. rugosa. Subcellular localizations of RrSIP1 (RrGT11) and RrGTγ-4 were on chloroplasts while RrGT-1 and RrSIP2 (RrGT14) located on cell nucleus. Regulation of ion transport could be the most important role of RrSIPs and RrGTγ-4. And RrGT-1 could be a halophytic gene with higher transcription abundance than glycophytic GT-1. These results provide key clue for further investigations of roles of RrGTs in salt stress response and would be helpful in the understanding the salt tolerance regulation mechanism of R. rugosa.

BOX38, a DNA Marker for Selection of Essential Oil Yield of Rosa × rugosa.

Rosa rugosa L. was a famous aromatic plant whose cultivars (Rosa × rugosa) have been widely used in the perfume industry in Asia. The perfume market looks for rose cultivars bearing higher essential oil, while the oil yields of most R. × rugosa have not been evaluated due to limiting conditions, such as insufficient cultivation areas. Here, we tested the yield and the aroma components of essential oil of 19 R. × rugosa. The results indicated that the yields of nerol, citronellol, and geraniol could represent an alternative index of the total yield of essential oil. Sequence syntenic analysis indicated that the Rosa genus specific cis-element Box38 was highly polymorphic. The Box38 region isolation of Rosa × rugosa by flanked primers proved that Box38 repeat number was significantly positively correlated with the essential oil yield of the corresponding cultivar. In the breeding of Rosa × rugosa, six-Box38-repeat could be a robust threshold for selection of high-essential-oil roses. Together, we found that Box38 was a DNA marker for essential oil yield and that it would be helpful in the early selection and breeding of essential oil roses.

Simple Sequence Repeat Fingerprint Identification of Essential-Oil-Bearing Rosa rugosa via High-Resolution Melting (HRM) Analysis.

Oil-bearing Rosa rugosa are popular in the essential oil and perfume markets. The similar botanical characteristics between high-oil-yield or low-oil-yield cultivars are confusing and it is hard for farmers or breeders to identify the high-oil-yield cultivar by phenotype difference. High-resolution melting (HRM) analysis of simple sequence repeats (SSRs) can construct accurate DNA fingerprints quickly, which was shown to be effective for identification of closely related cultivars of R. rugosa. Optimization of HRM-SSR indicated that the 10 µL HRM reaction mixture containing 20 ng of genomic DNA of R. rugosa and 0.75 µL of 10 µmol/L of each primer with an annealing temperature of 64 °C was a robust SSR genotyping protocol. Using this protocol, 9 polymorphic SSR markers with 3-9 genotypes among the 19 R. rugosa cultivars were identified. The top three polymorphic makers SSR9, SSR12 and SSR19 constructed a fingerprint of all cultivars, and the rare insertion in the flanking sequences of the repeat motif of SSR19 generated three characteristic genotypes of three high-oil-yield cultivars. These results may be economical and practical for the identification of high-oil-yield R. rugosa and be helpful for the selection and breeding of oil-bearing roses.

Genome-Wide Identification of LATERAL ORGAN BOUNDARIES DOMAIN (LBD) Transcription Factors and Screening of Salt Stress Candidates of Rosa rugosa Thunb.

LATERAL ORGAN BOUNDARIES DOMAIN (LBD) transcription factors are regulators of lateral organ morphogenesis, boundary establishment, and secondary metabolism in plants. The responsive role of LBD gene family in plant abiotic stress is emerging, whereas its salt stress responsive mechanism in Rosa spp. is still unclear. The wild plant of Rosa rugosa Thunb., which exhibits strong salt tolerance to stress, is an ideal material to explore the salt-responsive LBD genes. In our study, we identified 41 RrLBD genes based on the R. rugosa genome. According to phylogenetic analysis, all RrLBD genes were categorized into Classes I and II with conserved domains and motifs. The cis-acting element prediction revealed that the promoter regions of most RrLBD genes contain defense and stress responsiveness and plant hormone response elements. Gene expression patterns under salt stress indicated that RrLBD12c, RrLBD25, RrLBD39, and RrLBD40 may be potential regulators of salt stress signaling. Our analysis provides useful information on the evolution and development of RrLBD gene family and indicates that the candidate RrLBD genes are involved in salt stress signaling, laying a foundation for the exploration of the mechanism of LBD genes in regulating abiotic stress.

Retinal nerve fiber layer characteristics in patients with spontaneous intracranial hypotension: a retrospective case series.

OBJECTIVE: To evaluate changes in retinal nerve fiber layer (RNFL) and macular thicknesses, included ganglion cell-inner plexiform layer (GCIPL) thickness, in patients with spontaneous intracranial hypotension (SIH). METHODS: This was a retrospective, nonrandom, observational case series study. Comprehensive ophthalmic examinations and systemic examinations were performed. Spectral domain optical coherence tomography angiography scanning was used to measure peripapillary RNFL thickness and macular volume. RESULTS: In total, 108 eyes in 54 patients with SIH were evaluated; these were compared with 108 eyes in 54 healthy controls. The mean ages were 38.2 ± 9.4 years (patients with SIH) and 38.9 ± 9.4 years (healthy controls). In both groups, 33 patients were women (61.1%). The peripapillary RNFL and GCIPL were thinner in patients with SIH than in healthy controls (100.08 ± 9.94 µm vs 104.83 ± 8.35 µm and 81.46 ± 5.67 µm vs 85.67 ± 4.57 µm, respectively). Among patients with SIH, the GCIPL was thinner in patients with visual field defects (79.81 ± 5.62 µm vs 82.39 ± 5.12 µm). CONCLUSIONS: The RNFL and GCIPL were thinner in patients with SIH than in healthy controls. The GCIPL was thinner in eyes with visual field defects among patients with SIH.

Overexpression of a Rosa rugosa Thunb. NUDX gene enhances biosynthesis of scent volatiles in petunia.

Rosa rugosa is an important natural perfume plant in China. Rose essential oil is known as 'liquid gold' and has high economic and health values. Monoterpenes are the main fragrant components of R. rugosa flower and essential oil. In this study, a member of the hydrolase gene family RrNUDX1 was cloned from Chinese traditional R. rugosa 'Tang Hong'. Combined analysis of RrNUDX1 gene expression and the aroma components in different development stages and different parts of flower organ, we found that the main aroma component content was consistent with the gene expression pattern. The RrNUDX1 overexpressed Petunia hybrida was acquired via Agrobacterium-mediated genetic transformation systems. The blades of the transgenic petunias became wider and its growth vigor became strong with stronger fragrance. Gas chromatography with mass spectrometry analysis showed that the contents of the main aroma components of the transgenic petunias including methyl benzoate significantly increased. These findings indicate that the RrNUDX1 gene plays a role in enhancing the fragrance of petunia flowers, and they could lay an important foundation for the homeotic transformation of RrNUDX1 in R. rugosa for cultivating new R. rugosa varieties of high-yield and -quality essential oil.

Comparative analysis of headspace volatiles of Chinese Rosa rugosa.

The floral headspace compounds of Chinese Rosa rugosa germplasms that were isolated by an automated headspace sampler with built-in trap, and followed by gas chromatography-mass spectrometry for identification and quantification. Up to 33 volatile compounds were identified from the 23 rose germplasms, including nine alcohols, five esters, three alkanes, 10 terpenes, three aldehydes, two ketones, and one ether. The main floral components identified were 2-phenylethanol, ß-citronellol, ethanol, and n-hexane. 'xizi', 'miaofengshan', 'xiangciguo', and 'tangbai' contained the highest amounts of 2-phenylethanol at 84.66 µg·g⁻¹, ß-citronellol at 70.98 µg·g⁻¹, ethanol at 83.87 µg·g⁻¹, and n-hexane at 18.23 µg·g⁻¹, respectively. 'Rongchengyesheng', 'tanghong', 'xizi', 'miaofengshan', and 'baizizhi' could be considered good materials for extracting rose oil and breeding new cultivars.

Contribution of biogenic sources to secondary organic aerosol in the summertime in Shaanxi, China.

A revised Community Multi-scale Air Quality (CMAQ) model with updated secondary organic aerosol (SOA) yields and a more detailed description of SOA formation from isoprene (ISOP) oxidation was applied to study the spatial distribution of SOA, its components and precursors in Shaanxi in July of 2013. The emissions of biogenic volatile organic compounds (BVOCs) were generated using the Model of Emissions of Gases and Aerosols from Nature (MEGAN), of which ISOP and monoterpene (MONO) were the top two, with 1.73 × 109 mol and 1.82 × 108 mol, respectively. The spatial distribution of BVOCs emission was significantly correlated with the vegetation coverage distribution. ISOP and its intermediate semi-volatile gases were up to ∼7.0 and ∼1.4 ppb respectively in the ambient. SOA was generally 2-6 µg/m3, of which biogenic SOA (BSOA) accounted for as high as 84% on average. There were three main BVOCs Precursors including ISOP (58%) and MONO (8%) emit in the studied domain, and ISOP (9%) transported. The Guanzhong Plain had the highest BSOA concentrations of 3-5 µg/m3, and the North Shaanxi had the lowest of 2-3 µg/m3. More than half of BSOA was due to reactive surface uptake of ISOP epoxide (0.2-0.7 µg/m3, ∼19%), glyoxal (GLY) (0.2-0.5 µg/m3, ∼11%) and methylglyoxal (MGLY) (0.4-1.4 µg/m3, ∼32%), while the remaining was due to the traditional equilibrium partitioning of semi-volatile components (0.1-1.2 µg/m3, ∼25%) and oligomerization (0.2-0.4 µg/m3, ∼12%). Overall, SOA formed from ISOP contributed 1-3 µg/m3 (∼80%) to BSOA.

Mps1 is associated with the BRAFV600E mutation and predicts poor outcome in patients with colorectal cancer.

Colorectal cancer (CRC) with the V600E mutation of B-Raf proto-oncogene serine/threonine kinase (BRAFV600E) mutation is insensitive to chemotherapy and is indicative of a poor patient prognosis. Although BRAF inhibitors have a marked effect on malignant melanoma harboring the BRAFV600E mutation, they have a limited effect on patients with CRC with the same BRAF mutation. A previous study identified a novel gene, monopolar spindle protein kinase 1 (Mps1), a downstream target of BRAFV600E only, rather than of wild-type BRAF as well, which contributes to tumorigenesis in melanoma. In the present study, the incidence of BRAFV600E in patients with CRC was identified and the correlation of Mps1, phospho-extracellular-signal-regulated kinase (p-ERK) and BRAFV600E was investigated. The results indicated that the mutation rate of BRAFV600E was 5.2% in CRC. Poorly differentiated tumors and mucinous tumors have a significantly higher incidence of BRAFV600E compared with well-differentiated tumors and non-mucinous tumors (P<0.05). Kaplan-Meier survival analysis indicated that the survival rate was markedly lower in patients with BRAFV600E compared with in patients with wild-type BRAF (BRAFWT). The expression of p-ERK and Mps1 in CRC with BRAFV600E was significantly higher compared with in CRC with BRAFWT (P<0.05), and their expression is associated with cancer classification, degree of differentiation and lymph node transfusion (P<0.05). In addition p-ERK expression was positively correlated with Mps1 expression, with a contingency coefficient of 0.679 (P=0.002). In conclusion, the results of the present study indicated that Mps1 was significantly associated with BRAFV600E/p-ERK and may serve a crucial function in the development of CRC. The results of the present study raise the possibility that targeting the oncogenic BRAF and Mps1, particularly when in conjunction, could provide promising therapeutic opportunities for the treatment of CRC.

Improvement in Submergence Tolerance of Cherry Through Regulation of Carbohydrate Metabolism and Plant Growth by PsERF and PsCIPK.

Cherry is an important fruit tree with delicious taste and high economic value, which have been planted worldwide. However, this species cannot withstand the presence of excessive amount of water; submergence injury sometimes occurs during cultivation of cherry and results in severe economic losses. By using a submergence-tolerant germplasm Prunus serrulata "Yimeng" and a submergence-sensitive germplasm Prunus pseudocerasus "Aihua" as test materials, this study cloned PsERF and PsCIPK, which are related to submergence tolerance in cherry, and analyzed the expression of PsERF and PsCIPK in submergence-tolerant and submergence-sensitive germplasms under submergence stress; moreover, the consistency and correlation of such expression with carbohydrate metabolism and plant growth-related genes (PsPDC, PsSUS, PsRAMY, and PsEXP) were analyzed. The results showed that PsERF and PsCIPK influence the expression of PsPDC, PsSUS, PsRAMY, and PsEXP at different extents under submergence and during recovery to systematically improve the submergence resistance of P. serrulata "Yimeng". This study lays the important theoretical and practical foundation for molecular improvement and germplasm innovation in submergence tolerance in cherry through genetic engineering.

Cloning and Functional Verification of Genes Related to 2-Phenylethanol Biosynthesis in Rosa rugosa.

In China, Rosa rugosa is cultivated as a source of natural perfumes. Rose essential oil is known as "liquid gold", given its high economic and health value. 2-phenylethanol accounts for more than 10% of the total mass fraction of the essential oil derived from R. rugosa. The regulatory mechanisms underlying 2-phenylethanol metabolism in R. rugosa, however, remain unclear. In this study, RrAAAT and RrPPDC1, two genes related to 2-phenylethanol synthesis, were cloned from R. rugosa. Expression analysis revealed that RrAAAT and RrPPDC1 were highly expressed in rose flowers in the full opening and withering stages, and in calyxes. The overexpression vectors of RrAADC, RrAAAT, and RrPPDC1 were established and transformed into Petunia hybrida via Agrobacterium-mediated genetic transformation. Results demonstrated that the overexpression of RrAADC and RrAAAT increased the 2-phenylethanol content of transgenic petunia flowers. The results of this study provide a basis for the introduction of genes related to 2-phenylethanol synthesis into roses to increase the 2-phenylethanol content of rose essential oil.

Effects of miR-144 on the sensitivity of human anaplastic thyroid carcinoma cells to cisplatin by autophagy regulation.

BACKGROUND: We investigated the influence of miR-144 on the cisplatin-sensitivity of anaplastic thyroid carcinoma (ATC) cells and explored the internal molecular mechanism of miR-144. METHODS: Thyroid cancer cells ARO, TPC1 and normal thyroid cells HT-ori3 were used in this research. Expressions of miR-144 and TGF-α were uncovered by western blot and qRT-PCR. Expressions of autophagy-related protein LC3 II and apoptosis-related protein Caspase-3 and PARP were explored by western blot and immunofluorescence. Cell viability was detected by MTT assay and apoptosis condition was revealed by flow cytometric analysis and TUNEL staining. Dual-luciferase reporter assay was employed to verify the target relationship. Tissue sections were detected by IHC. Xenograft assay was conducted to further verify conclusions in vivo. RESULTS: MiR-144, which was low expressed in ATC cells and tissues, could inhibit autophagy activation induced by cisplatin, enhancing the sensitivity of ATC cells to cisplatin, and promoting cell apoptosis. TGF-α was the target of miR-144 and was negatively regulated by it. MiR-144 could improve the sensitivity of ATC cells to cisplatin and inhibit tumor growth by suppressing TGF-α both in vitro and in vivo. CONCLUSION: MiR-144 could inhibit autophagy of ATC cells by down-regulating TGF-α, enhancing the cisplatin-sensitivity of ATC cells.