The impact of internet health information seeking on COVID-19 vaccination behavior in China.

BACKGROUND: Amid the COVID-19 pandemic, the significance of vaccination has been emphatically underscored. As the foremost and pivotal measure for preventing COVID-19 transmission, the COVID-19 pneumonia vaccine plays an instrumental role in the global response to this infectious respiratory disease. However, COVID-19 vaccination coverage remains challenging in low- and middle-income countries and regions. Studies have found that frequent seeking to health information is also associated with healthier behaviors, but these studies have not yet focused on the impact of Internet health information seeking on individual vaccination and the mechanism of this effect. METHODS: Data were obtained from the Chinese General Social Survey (CGSS) conducted in 2021, which included 7,218 individual samples. This study used COVID-19 vaccination as indicators for the health of interest, the key independent variable is Internet health information seeking. This study tried to analyze the impact of Internet health information seeking on COVID-19 vaccination using an OLS model and PSM method. RESULTS: The results show that Internet health information seeking has a significant positive impact on COVID-19 vaccination. This result passed a series of robustness tests. The mechanism analysis indicated that compared to non-Internet health information seeking individuals, Internet health information seeking individuals could know the superior advantages of vaccination and the potential for immunization through this method. And individuals who use Internet to seeking health information are more likely to acknowledge the constructive impact of online information on health behavior. This helps to explain why Internet health information seeking individuals have a higher rate of COVID-19 vaccination. CONCLUSIONS: This study delves into the influence of Internet health information seeking on individual COVID-19 vaccination within the digital era. The outcomes underscore that Internet-mediated seeking vaccine information holds the potential to bolster individuals' comprehension of vaccination benefits and foster increased acceptance of such information.

Efficacy and safety of erector spinae plane block for postoperative analgesia in breast cancer surgery-A systematic review and meta-analysis.

BACKGROUND AND OBJECTIVES: We aim to evaluate the analgesic efficacy and safety of erector spinae plane block (ESPB) for postoperative analgesia in breast cancer surgeries. METHODS: PubMed, Web of Science, CBM, Embase, Cochrane, Wanfang, VIPP, and CNKI were searched to identify published eligible randomized controlled trials. The primary results were the postoperative 24 h morphine consumption and pain scores, while the secondary outcomes included pain scores at other times, press times of patient-controlled intravenous analgesia (PCIA), times to request for first rescue analgesia, the incidence of request for rescue analgesia, opioid-related complications, nerve blocks related complications and patient satisfaction. RESULTS: We included 20 studies meeting the inclusion criteria, which involved 1293 participants. The morphine consumption and the pain scores during 24 h postoperatively were significantly decreased in the ESPB group versus the control group (p < 0.00001). Furthermore, ESPB also reduced pain scores at other time points, press times of PCIA, and times to first rescue analgesia requirement. Meanwhile, there was a lower incidence of postoperative nausea and vomiting, and skin pruritus in the ESPB group than in the control group. CONCLUSIONS: Compared to general anesthesia alone, ESPB combined with general anesthesia can effectively reduce the postoperative pain intensity within 48 h and opioid consumption within 24 h after breast cancer surgery, and reduce the incidence of opioid and nerve blocks related complications.

Gender differences in vision health-seeking behavior and vision health outcomes among rural Chinese schoolchildren by birth order and family size.

BACKGROUND: The gender gap remains a major impediment in the path toward equality, and it is especially wide in low-income countries. Gender differences in health-seeking behaviors may be a factor. Family size and childbirth order are two critical factors affecting family resource allocation. This study examines gender differences in healthcare-seeking behaviors among children with visual impairment in rural China across different family structures (birth order and family size). METHODS: We draw on a dataset containing 19,934 observations constructed by combining data from 252 different school-level surveys spanning two provinces. The surveys were all conducted in 2012 using uniform survey instruments and data collection protocols in randomly selected schools across western provinces in rural China. The sample children range in grades from 4 to 5. Our analysis compares rural girls with rural boys regarding vision health outcome and behavior (vision examination and correction). RESULTS: The findings revealed that girls have worse vision than boys. Regarding vision health behaviors, girls have a lower overall vision examination rate than boys. There is no gender difference when the sample student is the only child or the youngest child in the family, but there is still a gender difference when the sample student is the oldest child in the family or the middle child in the birth order. When it comes to vision correction behavior, boys are more likely to own eyeglasses than girls are for groups of students with mild visual impairment, even when the sample student is the only child in the family. However, when the sample student has another brother or sister (the sample student is the youngest, the oldest child in the family, or the middle child in the birth order), the gender difference disappears. CONCLUSIONS: Gender differences in vision health outcomes are correlated with gender differences in vision health-seeking behaviors among rural children. Depending on the birth order and family size, gender disparities in visual health practices vary. In the future, consideration should be given to providing medical subsidies to reduce the cost of vision health behaviors and to provide information interventions to change gender inequality in households and promote equality in children's vision health behaviors. TRIAL REGISTRATION: The trial was approved by the Stanford University Institutional Review Board (Protocol No. ISRCTN03252665). Permission was received from local Boards of Education in each region and the principals of all schools. The principles of the Declaration of Helsinki were followed throughout. Written informed consent was obtained from at least one parent for all child participants.

The impact of parental migration on left-behind children's vision health in rural China.

BACKGROUND: Parental migration is an important factor affecting left-behind children's health. However, few studies have addressed the effect of parental migration on children's vision health in China. To fill the gap, this study aimed to assess the impact of parental migration on left-behind children's vision health and to explore the possible mechanisms of the effect. METHODS: Data were obtained from the baseline survey of the China Education Panel Survey (CEPS), which included over 10,000 junior high school students. This study used myopia, the most common vision problem among junior high school students, and tried to analyze whether myopia was corrected with eyeglasses as indicator variables of vision health. The impact of parental migration on vision health was assessed using an instrumental variables approach. RESULTS: The results show that parental migration reduced the likelihood of myopia in left-behind children and decreased the possibility of myopic left-behind children being corrected. This result passed a series of robustness tests. The mechanism analysis indicated that compared to non-left-behind children, left-behind children spent more time on outdoor activities and less time on after-school classes, reducing their risk of being myopic. Further, because left-behind children live apart from their parents, their myopia problem is more difficult for parents to notice, and left-behind children are less likely to inform their parents of their myopia than non-left-behind children actively. This helps to explain why left-behind children have a lower correction rate with eyeglasses. CONCLUSIONS: Our findings suggest that parental migration, while not increasing the prevalence of myopia in left-behind children, has led to inequity in myopic left-behind children's correction. Given the severe consequences of uncorrected myopia, action is required to enhance the correction rate of myopic left-behind children.

A Comprehensive Investigation on Multi-User Interference Effects in Vehicular Visible Light Communications.

Vehicular visible light communications (VLC) are considered a suitable technology for vehicular platooning applications. Nevertheless, this domain imposes strict performance requirements. Although numerous works have shown that VLC technology is compatible with platooning applications, existing studies are mainly focused on the physical layer performances, mostly ignoring the disruptive effects generated by neighboring vehicular VLC links. Nevertheless, the 5.9 GHz Dedicated Short Range Communications (DSRC) experience has shown that mutual interference can significantly affect the packed delivery ratio, pointing out that these effects should be analyzed for vehicular VLC networks as well. In this context, this article provides a comprehensive investigation focused on the effects of mutual interference generated by neighboring vehicle-to-vehicle (V2V) VLC links. Therefore, this work provides an intensive analytical investigation based on simulation and also on experimental results that demonstrate that although ignored, the influence of mutual interference is highly disruptive in vehicular VLC applications. Hence, it has been shown that without preventive measures, the Packet Delivery Ratio (PDR) can decrease below the imposed 90% limit for almost the entire service area. The results have also shown that although less aggressive, multi-user interference affects V2V links even in short-distance conditions. Therefore, this article has the merit of emphasizing a new challenge for vehicular VLC links and points out the importance of multiple-access techniques integration.

Addressing Multi-User Interference in Vehicular Visible Light Communications: A Brief Survey and an Evaluation of Optical CDMA MAC Utilization in a Multi-Lane Scenario.

Visible Light Communications (VLC) are developing as an omnipresent solution for inter-vehicle communications. Based on intensive research efforts, the performance of vehicular VLC systems has significantly improved in terms of noise resilience, communication range, and latencies. Nevertheless, in order to be ready for deployment in real applications, solutions for Medium Access Control (MAC) are also required. In this context, this article provides an intensive evaluation of several optical CDMA MAC solutions and of their efficiency in mitigating the effect of Multiple User Interference (MUI). Intensive simulation results showed that an adequately designed MAC layer can significantly reduce the effects of MUI, ensuring an adequate Packet Delivery Ratio (PDR). The simulation results showed that based on the use of optical CDMA codes, the PDR can be improved from values as low as 20% up to values between 93.2% and 100%. Consequently, the results provided in this article show the high potential of optical CDMA MAC solutions in vehicular VLC applications, reconfirm the high potential of the VLC technology in inter-vehicle communications, and emphasize the need to further develop MAC solutions designed for such applications.

Glucose-dependent insulinotropic polypeptide modifies adipose plasticity and promotes beige adipogenesis of human omental adipose-derived stem cells.

The adipocyte precursors (APs) located in white adipose tissue (WAT) are functionally significant in adipose plasticity and browning. Modifying adipogenesis or WAT browning targeted on APs is a promising mechanism for anti-obesity drug. We herein explored the in vitro actions and mechanisms of glucose-dependent insulinotropic polypeptide (GIP), a gut-derived peptide, in human adipose-derived mesenchymal stem cells (hADSCs) isolated from omentum. The hADSCs were cotreated with 100 nM GIP with or without equimolar concentration of GIP3-42 (a GIP receptor antagonist), and subsequently examined in vitro. CCK-8, EdU incorporation, and flow cytometry assays were used to assess cellular proliferation. Annexin V FTIC/PI double stain, TUNEL staining, and Western blot were applied for apoptosis evaluation. Adipogenesis was reflected by Western blot, real-time PCR, Oil Red O staining, mitochondrial staining, and mitochondrial DNA analysis. Results showed that GIP promoted proliferation and inhibited apoptosis of hADSCs via pleiotropic effects. Besides, GIP facilitated de novo beige adipogenesis, by accelerating mitotic clonal expansion (MCE), upregulating core adipogenic regulators (C/EBPα and PPARγ), augmenting beige-related genes (UCP1, PGC1α, and PRDM16), increasing mitochondrial content and improving beige adipocyte functionalities. Above all, our study expands knowledge on the mechanisms of GIP modifying adipogenesis especially in inducing beige adipogenesis, and thus provides a theoretical support for clinical usage of GIP on obesity treatment.

Long noncoding RNA LINC00673-v4 promotes aggressiveness of lung adenocarcinoma via activating WNT/ß-catenin signaling.

It is well recognized that metastasis can occur early in the course of lung adenocarcinoma (LAD) development, and yet the molecular mechanisms driving this capability of rapid metastasis remain incompletely understood. Here we reported that a long noncoding RNA, LINC00673, was up-regulated in LAD cells. Of note, we first found that LINC00673-v4 was the most abundant transcript of LINC00673 in LAD cells and its expression was associated with adverse clinical outcome of LAD. In vitro and in vivo experiments demonstrated that LINC00673-v4 enhanced invasiveness, migration, and metastasis of LAD cells. Mechanistically, LINC00673-v4 augmented the interaction between DDX3 and CK1ε and thus the phosphorylation of dishevelled, which subsequently activated WNT/ß-catenin signaling and consequently caused aggressiveness of LAD. Antagonizing LINC00673-v4 suppressed LAD metastasis in vivo. Together, our data suggest that LINC00673-v4 is a driver molecule for metastasis via constitutively activating WNT/ß-catenin signaling in LAD and may represent a potential therapeutic target against the metastasis of LAD.

SLC6A15 acts as a tumor suppressor to inhibit migration and invasion in human papillary thyroid cancer.

Solute Carrier Family 6 Member 15 (SLC6A15), a sodium-dependent neutral amino acid transporter, has been found with dysregulated expression in several kinds of cancers. However, the expression pattern and the biological functions of SLC6A15 in papillary thyroid cancer (PTC) remain unknown. In this study, we found that SLC6A15 was downregulated in PTC, which was related to N classification. Ectopic overexpression of SLC6A15 impaired migratory and invasive abilities of PTC cell in vitro. In addition, we identified intercellular adhesion molecule-1, a vital oncogene in thyroid cancer progression, was involved in the effects of SLC6A15 on PTC cell. These results indicate that SLC6A15 acts as a tumor suppressor and might be a potential therapeutic target in the treatment of PTC.

Polyoxometalates-Based Semi-flexible Metal-Semiconductor Triboelectric Nanogenerators for Low Frequency and Small Amplitude Mechanical Energy Harvesting.

The development of high-performance and low-cost durable triboelectric nanogenerators (TENGs) is essential for converting mechanical energy into electrical energy. Many organic polymer friction materials used widely have thermal stability problems, which makes TENGs with semiconductors as friction materials stand out. Here, we report a semi-flexible TENG based on metal and TiO2 modified by polyoxometalates (POMs) as pure inorganic friction materials. Six different POMs are firstly selected to modify the friction materials of TENGs, and the output performance of TENGs with different POMs-modified semiconductors and different metals as friction materials are tested. Compared with the unmodified TENGs, the open-circuit voltage (VOC ) of the optimal Ag-K6 P2 Mo18 O62 (P2 Mo18 )/TiO2 TENG device is increased by more than 4 times, which is mainly attributed to the strong electron-accepting and storage capabilities of POMs. This study has demonstrated that TENGs modified by POMs have potential application prospects and provided a new method for increasing the electrical output of TENGs.

Enhanced terahertz focusing for a graphene-enabled active metalens.

Graphene-based terahertz (THz) metasurfaces have the advantages of ultra-small thickness, electrical tunability, and fast tuning speed. However, many such structures suffer low efficiency, especially for transmissive devices. Here we propose a hybrid structure for focusing THz waves with tunability and enhanced focusing efficiency, which is composed of a graphene-loaded metallic metasurface sandwiched by two mutually orthogonal gratings. Experimental results show that due to the multi-reflection between the metasurface layer and the grating layer, the focusing efficiency is enhanced by 1.8 times, and the focal length of the metalens is increased by 0.61 mm when the applied gate voltage on the graphene is increased from 0 V to 1.4 V. We hope the proposed structure may open a new avenue for reconfigurable THz metasurfaces with high efficiencies.

Identification and functional characterization of diterpene synthases for triptolide biosynthesis from Tripterygium wilfordii.

Tripterygium wilfordii, which has long been used as a medicinal plant, exhibits impressive and effective anti-inflammatory, immunosuppressive and anti-tumor activities. The main active ingredients are diterpenoids and triterpenoids, such as triptolide and celastrol, respectively. A major challenge to harnessing these natural products is that they are found in very low amounts in planta. Access has been further limited by the lack of knowledge regarding their underlying biosynthetic pathways, particularly for the abeo-abietane tri-epoxide lactone triptolide. Here suspension cell cultures of T. wilfordii were found to produce triptolide in an inducible fashion, with feeding studies indicating that miltiradiene is the relevant abietane olefin precursor. Subsequently, transcriptome data were used to identify eight putative (di)terpene synthases that were then characterized for their potential involvement in triptolide biosynthesis. This included not only biochemical studies which revealed the expected presence of class II diterpene cyclases that produce the intermediate copalyl diphosphate (CPP), along with the more surprising finding of an atypical class I (di)terpene synthase that acts on CPP to produce the abietane olefin miltiradiene, but also their subcellular localization and, critically, genetic analysis. In particular, RNA interference targeting either both of the CPP synthases, TwTPS7v2 and TwTPS9v2, or the subsequently acting miltiradiene synthase, TwTPS27v2, led to decreased production of triptolide. Importantly, these results then both confirm that miltiradiene is the relevant precursor and the relevance of the identified diterpene synthases, enabling future studies of the biosynthesis of this important bioactive natural product.

Probing the function of protein farnesyltransferase in Tripterygium wilfordii.

KEY MESSAGE: We found two subunits FTase/GGTaseI-α and FTase-ß formed a heterodimer to transfer a farnesyl group from FPP to protein N-dansyl-GCVLS, confirming they are responsible for protein farnesylation in planta. Tripterygium wilfordii is a medicinal plant with a broad spectrum of anti-inflammatory, immunosuppressive and anti-cancer activities. Recently, a number of studies have focused on investigating the biosynthetic pathways of its bioactive compounds, whereas little attention has been paid to the enzymes which play important roles in regulating diverse developmental processes of T. wilfordii. In this study, we report for the first time the identification and characterization of two subunits of farnesyltransferase (FTase), farnesyltransferase/geranylgeranyltransferase I-α (TwFTase/GGTase I-α) and farnesyltransferase-ß (TwFTase-ß), in this important medicinal plant. Cell-free in vivo assays, yeast two-hybrid (Y2H) and pull-down assays showed that the two subunits interact with each other to form a heterodimer to perform the role of specifically transferring a farnesyl group from FPP to the CAAX-box protein N-dansyl-GCVLS. Furthermore, we discovered that the two subunits had the same cytoplasmic localization pattern and displayed the same tissue expression pattern. These results indicated that we identified a functional TwFTase enzyme which contains two functionally complementary subunits TwFTase/GGTase I-α and TwFTase-ß, which provides us promising genetic targets to construct transgenic plants or screen for more adaptable T. wilfordii mutants, which are able to survive in changing environments.

Eudesmane-type sesquiterpene diols directly synthesized by a sesquiterpene cyclase in Tripterygium wilfordii.

Cryptomeridiol, a typical eudesmane diol, is the active principle component of the antispasmodic Proximol. Although it has been used for many years, the biosynthesis pathway of cryptomeridiol has remained blur. Among terpenoid natural products, terpenoid cyclases are responsible for cyclization and generation of hydrocarbon backbones. The cyclization is mediated by carbocationic cascades and ultimately terminated via deprotonation or nucleophilic capture. Isoprene precursors are, respectively, converted into hydrocarbons or hydroxylated backbones. A sesquiterpene cyclase in Tripterygium wilfordii (TwCS) was determined to directly catalyze (E,E)-farnesyl pyrophosphate (FPP) to unexpected eudesmane diols, primarily cryptomeridiol. The function of TwCS was characterized by a modular pathway engineering system in Saccharomyces cerevisiae The major product determined by NMR spectroscopy turned out to be cryptomeridiol. This unprecedented production was further investigated in vitro, which verified that TwCS can directly produce eudesmane diols from FPP. Some key residues for TwCS catalysis were screened depending on the molecular model of TwCS and mutagenesis studies. As cryptomeridiol showed a small amount of volatile and medicinal properties, the biosynthesis of cryptomeridiol was reconstructed in S. cerevisiae Optimized assays including modular pathway engineering and the CRISPR-cas9 system were successfully used to improve the yield of cryptomeridiol in the S. cerevisiae The best engineered strain TE9 (BY4741 erg9::Δ-200-176 rox1::mut/pYX212-IDI + TwCS/p424-tHMG1) ultimately produced 19.73 mg/l cryptomeridiol in a shake flask culture.

Exendin-4 modifies adipogenesis of human adipose-derived stromal cells isolated from omentum through multiple mechanisms.

BACKGROUND: Abdominal obesity is considered a major factor in the development of metabolic disorders. Glucagon-like peptide-1 (GLP-1) has been reported to have positive effects on improving body metabolism and to reducing insulin resistance. However, it remains less clear whether GLP-1 plays a role in the adipogenesis process of visceral fat. METHODS: Here, we analyzed the in vitro actions and probable mechanisms of Exendin-4, a GLP-1 receptor agonist, on human adipose-derived stromal cells (hADSCs) isolated from omentum. RESULTS: Our results demonstrated that Exendin-4 improved cell viability via promoting proliferation and inhibiting apoptosis in hADSCs isolated from omentum. Mechanistically, the activation of MAPK/ ERK1/2, Akt/GSK-3ß, and PKA/CREB pathways and downstream consequences induced are involved in the proliferative and anti-apoptotic roles of Exendin-4. More intriguingly, Exendin-4 could promote the differentiation of omental hADSCs. Underlying mechanisms of the differentiation of hADSCs are associated with the upregulation of the expression of pro-adipogenic genes and downregulation of the expression of anti-adipogenic genes. CONCLUSION: Our data demonstrate that Exendin-4 modifies adipogenesis of hADSCs isolated from omentum through multiple mechanisms, these effects could contribute to the protective actions of GLP-1 receptor agonist body metabolism and insulin sensitivity.

Cloning and functional analysis of two sterol-C24-methyltransferase 1 (SMT1) genes from Paris polyphylla.

The biosynthetic pathways of phytosterols and steroidal saponins are located in two adjacent branches which share cycloartenol as substrate. The rate-limiting enzyme S-adenosyl-L-methionine-sterol-C24-methyltransferase 1 (SMT1) facilitates the metabolic flux toward phytosterols. It catalyzes the methylation of the cycloartenol in the side chain of the C24-alkyl group, to generate 24(28)-methylene cycloartenol. In this study, we obtained two full-length sequences of SMT1 genes from Pari polyphylla, designated PpSMT1-1 and PpSMT1-2. The full-length cDNA of PpSMT1-1 was 1369 bp long with an open reading frame (ORF) of 1038 bp, while the PpSMT1-2 had a length of 1222 bp, with a 1005 bp ORF. Bioinformatics analysis confirmed that the two cloned SMTs belong to the SMT1 family. The predicted function was further validated by performing in vitro enzymatic reactions, and the results showed that PpSMT1-1 encodes a cycloartenol-C24-methyltransferase, which catalyzes the conversion of cycloartenol to 24-methylene cycloartenol, whereas PpSMT1-2 lacked this catalytic activity. The tissue expression patterns of the two SMTs revealed differential expression in different organs of Paris polyphylla plants of different developmental stage and age. These results lay the foundation for detailed genetic studies of the biosynthetic pathways of steroid compounds, which constitute the main class of active substances found in P. polyphylla.

Experimental comparison of pulse-amplitude and spatial modulations for vehicle-to-vehicle visible light communication in platoon configurations.

Visible light communication (VLC) is an attractive complementary communication technology for vehicular applications such as platooning. Although data rates around 100 kbps are enough for crucial data transmission, it may be useful to reach a few megabits per second for other applications like networking. Such data rates can be reached by using appropriate modulations and clock rates. In this paper, three forms of pulse amplitude modulations (PAM) are compared in a vehicular context: on-off keying (OOK), PAM-4 and generalized space-shift keying (GSSK). A prototype based on off-the-shelf light-emitting diodes (LED) headlamps is used for static tests in straight line configuration, with an inter-vehicle distance up to 30 m, and curves of minimum radius 100 m and inter-vehicle distance of 10 m. These tests show that OOK and GSSK are the most interesting modulations for highway platooning applications. OOK provides indeed a good mobility while remaining simple to implement. A 1 Mbps link of BER below 10-6 is thus demonstrated. In GSSK, the data rate reaches 2 Mbps for an equivalent BER. These performances are obtained by using, in particular, two spatially distinct receivers, which limits strongly the complexity of GSSK decoding.

Trop2 enhances invasion of thyroid cancer by inducing MMP2 through ERK and JNK pathways.

BACKGROUND: Mounting evidence has showed that Tumor-associated calcium signal transducer 2 (Trop2) is upregulated in various kinds of human cancers and plays important roles in tumorigenesis. However, the expression status and functional significance of Trop2 in thyroid cancer are largely unknown. METHODS: We first determined the expression of Trop2 by using RNAseqV2 data sets for thyroid cancer deposited on The Cancer Genome Atlas (TCGA) website. The expression of Trop2 was then confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry assays. Cell invasion and migration were assessed by conducting Transwell and wound healing assays. Furthermore, we explored the underlying mechanisms by using real-time RT-PCR, Western blot, zymography, and luciferase reporter assays. RESULTS: In this study, we demonstrated that the expression of Trop2 was significantly elevated in thyroid cancer and that its expression level was correlated with the tumor-node-metastasis (TNM) staging and N classification. Dysregulation of Trop2 altered the invasive capability of thyroid cancer cells. Further mechanistic study revealed that MMP2 expression was upregulated by Trop2. Moreover, we found that the effects of Trop2 were dependent on ERK and JNK pathways. The results from clinical specimens showed that Trop2 expression correlated with MMP2 expression in primary thyroid cancer. CONCLUSION: The current study suggests that elevated expression of Trop2 may represent an important molecular hallmark that is biologically and clinically relevant to the progression of thyroid cancer.

[Cloning and expression analysis of kaurenoic acid oxidase gene in Tripterygium wilfordii].

Kaurenoic acid oxidase involved in biosynthesis pathway of gibberellin. According to the transcriptome database, the specific primers were designed and used in cloning the full-length cDNA of TwKAO, the bioinformatic analysis of the sequence was performed. The qRT-PCR were used to detect the expression level of TwKAO after MeJA treatment.The full-length cDNA of the TwKAO was 1 874 bp encoding a polypeptide of 487 amino acids.The calculate molecular weight was about 56.02 kDa,and the theoretical isoelectric point (pI) was 8.89. The relative expression level of TwKAO was deduced by MeJA and reached the highest at 12 h after the treatment.Plant tissue expression analysis indicated that, TwKAO expressed the highest in leaves,while lowest in roots.For the first time, we cloned and analyzed the expression characteristics of TwKAO, which laid a foundation for deep analysis of growing development and terpenoid secondary metabolites in T. wilfordii.

[Cloning and protein expression analysis of monoterpene synthase gene TwMS in Tripterygium wilfordii].

In this study, we cloned a monoterpene synthases, TwMS from Tripterygium wilfordii suspension cells. TwMS gene contained a 1 797 bp open reading frame (ORF), encoding a polypeptide of 579 amino acids, which deduced isoelectric point (pI) was 6.10 and the calculated molecular weight was 69.75 kDa. Bioinformation analysis showed that the sequence of TwMS was consistent with the feature of monoterpene synthases. Differential expression analysis revealed that the relative expression level of TwMS increased significantly after being induced by methyl jasmonate (MeJA). The highest expression level occurred at 24 h. TwMS protein was successfully expressed in Escherichia coli BL21 (DE3), which laid the foundation for identifying the function of T. wilfordii monoterpene synthases.