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To explore the relationship between soil nutrients, plant nutrients, and the growth and development of Trichosanthes kirilowii, the soil pH, organic matter, available nitrogen, available phosphorus, available potassium content and lea total nitrogen, total phosphorus, total potassium, and SPAD in different growth stages of T. kirilowii in the main production area of Shisliou City were measured and analyzed. The changes in soil nutrient content and lea nutrient content at different growth stages of T. kirilowii were compared, and correlation analysis was conducted. The results showed that the average soil pH, organic matter content alkaline nitrogen content available phosphorus content and available potassium content during the entire growth period of T. kirilowii were 7.03, 14.01 g/kg, 98.79 mg/kg, 14.84 mg/kg, and 135.20 mg/kg, respectively; the average total nitrogen content total phosphorus content total potassium content and SPAD of the leaves were 0.55%, 0.23%, 1.78%, and 77.66, respectively. The nutrient dynamics of T. kirilowii at different growth stages exhibited certain regularity, with most nutrients reaching their maximum values during the flowering and fruiting stages, and then showing a decreasing or stabilizing trend. There was a varying degree of correlation between the nutrient content of leaves and soil, among which the nitrogen, phosphorus, and potassium contents of leaves were significantly or extremely significantly correlated with soil organic matter and alkaline nitrogen content. It can be seen that the nutrient abundance or deficiency level of soil in T. kirilowii field significantly affected the nutrient content of the leaves at different growth stages, thereby restricting its growth and development status. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii (T. kirilowii) is a valuable plant used for both medicinal and edible purposes. It belongs to the Cucurbitaceae family. However, its phylogenetic position and relatives have been difficult to accurately determine due to the lack of mitochondrial genomic information. This limitation has been an obstacle to the potential applications of T. kirilowii in various fields. To address this issue, Illumina and Nanopore HiFi sequencing were used to assemble the mitogenome of T. kirilowii into two circular molecules with sizes of 245,700 bp and 107,049 bp, forming a unique multi-branched structure. The mitogenome contains 61 genes, including 38 protein-coding genes (PCGs), 20 tRNAs, and three rRNAs. Within the 38 PCGs of the T. kirilowii mitochondrial genome, 518 potential RNA editing sites were identified. The study also revealed the presence of 15 homologous fragments that span both the chloroplast and mitochondrial genomes. The phylogenetic analysis strongly supports that T. kirilowii belongs to the Cucurbitaceae family and is closely related to Luffa. Collinearity analysis of five Cucurbitaceae mitogenomes shows a high degree of structural variability. Interestingly, four genes, namely atp1, ccmFC, ccmFN, and matR, played significant roles in the evolution of T. kirilowii through selection pressure analysis. The comparative analysis of the T. kirilowii mitogenome not only sheds light on its functional and structural features but also provides essential information for genetic studies of the genus of Cucurbitaceae. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii (T. kirilowii) is a valuable plant used for both medicinal and edible purposes. It belongs to the Cucurbitaceae family. However, its phylogenetic position and relatives have been difficult to accurately determine due to the lack of mitochondrial genomic information. This limitation has been an obstacle to the potential applications of T. kirilowii in various fields. To address this issue, Illumina and Nanopore HiFi sequencing were used to assemble the mitogenome of T. kirilowii into two circular molecules with sizes of 245,700 bp and 107,049 bp, forming a unique multi-branched structure. The mitogenome contains 61 genes, including 38 protein-coding genes (PCGs), 20 tRNAs, and three rRNAs. Within the 38 PCGs of the T. kirilowii mitochondrial genome, 518 potential RNA editing sites were identified. The study also revealed the presence of 15 homologous fragments that span both the chloroplast and mitochondrial genomes. The phylogenetic analysis strongly supports that T. kirilowii belongs to the Cucurbitaceae family and is closely related to Luffa. Collinearity analysis of five Cucurbitaceae mitogenomes shows a high degree of structural variability. Interestingly, four genes, namely atp1, ccmFC, ccmFN, and matR, played significant roles in the evolution of T. kirilowii through selection pressure analysis. The comparative analysis of the T. kirilowii mitogenome not only sheds light on its functional and structural features but also provides essential information for genetic studies of the genus of Cucurbitaceae.

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Abstract Background The APETALA 2/ ethylene-responsive element binding factors (AP2/ERF), are thought to be associated with plant abiotic stress response, and involved in some plant hormone signaling pathways. Trichosanthes kirilowii is an important edible and medicinal crop, so far no research has been conducted on the TkAP2/ERF genes. Result In this study, a total of 135 TkERFs were identified, these genes were divided into 4 subfamilies and clustered into 13 groups. Moreover, 37 paralogous pairs were identified, with only two having Ka/Ks values greater than 1, proving that most TkERF genes underwent purifying selection during evolution. Co-expression networks constructed using transcriptome data at various flowering stages revealed that 50, 64, and 67 AP2/ERF genes correlated with members of the ethylene, gibberellin, and abscisic acid signaling pathways, respectively. When tissue cultured seedlings were treated with ETH, GA3 and ABA, 11, 12 and 17 genes were found to be up-regulated, respectively, suggesting that some members of the TkERF gene family may be involved in plant hormone signaling pathways. And under 4 ℃, PEG and NaCl treatment, 15, 20 and 19 genes were up-regulated, respectively, this suggested that these selected genes might be involved in plant abiotic stresses. Conclusions Overall, we identified 135 AP2/ERF family members, a comprehensive analysis of AP2/ERF gene expression patterns by RNA-seq and qRT-PCR showed that they played important roles in flower development and abiotic stress. This study provided a theoretical basis for the functional study of TkAP2/ERF genes and the genetic improvement of T. kirilowii.

Background: The APETALA 2/ ethylene-responsive element binding factors (AP2/ERF), are thought to be associated with plant abiotic stress response, and involved in some plant hormone signaling pathways. Trichosanthes kirilowii is an important edible and medicinal crop, so far no research has been conducted on the TkAP2/ERF genes. Result: In this study, a total of 135 TkERFs were identified, these genes were divided into 4 subfamilies and clustered into 13 groups. Moreover, 37 paralogous pairs were identified, with only two having Ka/Ks values greater than 1, proving that most TkERF genes underwent purifying selection during evolution. Co-expression networks constructed using transcriptome data at various flowering stages revealed that 50, 64, and 67 AP2/ERF genes correlated with members of the ethylene, gibberellin, and abscisic acid signaling pathways, respectively. When tissue cultured seedlings were treated with ETH, GA3 and ABA, 11, 12 and 17 genes were found to be up-regulated, respectively, suggesting that some members of the TkERF gene family may be involved in plant hormone signaling pathways. And under 4 ℃, PEG and NaCl treatment, 15, 20 and 19 genes were up-regulated, respectively, this suggested that these selected genes might be involved in plant abiotic stresses. Conclusions: Overall, we identified 135 AP2/ERF family members, a comprehensive analysis of AP2/ERF gene expression patterns by RNA-seq and qRT-PCR showed that they played important roles in flower development and abiotic stress. This study provided a theoretical basis for the functional study of TkAP2/ERF genes and the genetic improvement of T. kirilowii. [ABSTRACT FROM AUTHOR]

Copyright of Journal of South China Agricultural University is the property of Gai Kan Bian Wei Hui and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction: Ulcerative colitis (UC) is chronic inflammatory ailment that distinguished by diarrhea, stomach pain, reduced body weight, and hematochezia. UC affects nearly 400 per 100,000 peoples worldwide. In this exploration, we intended to fabricate the copper oxide nanoparticles from the T. kirilowii (TK-CuNPs) and inspect their curative potential against the Dextran sulfate disodium salt (DSS)-provoked colitis in animals. Methods: Formulated TK-CuNPs were characterized by UV–vis spectroscopy, FT-IR, SEM, TEM, and DLS techniques. UC was provoked in mice through administering of 2% DSS for 7-days and treated with the 10 mg/kg bodyweight of TK-CuNPs via intragastric intubation. Myeloperoxidase activity, colon weight, and length, and spleen weight were determined by standard methods. Pro-inflammatory mediators, i.e. IL-6, IL-1β, and TNF-α were inspected by the assay kits. The expression of COX-2 and iNOS were analyzed by immunohistochemistry. Histopathological changes in colon tissues were investigated microscopically by H&E staining. The cytotoxicity and in vitro anti-inflammatory activities of TK-CuNPs were analyzed in RAW-264.7 cells. Results: The outcomes of various characterization studies demonstrated the formation of TK-CuNPs. The TK-CuNPs supplementation to the DSS induced colitis mice demonstrated the appreciable bodyweight gain and reduced the DAI scores. TK-CuNPs improved the colon length and spleen weight and suppressed the MPO activity. The status of pro-inflammatory mediators IL-6, IL-1β, and TNF-α was appreciably suppressed by the TK-CuNPs treatment. The expressions of COX-2 and iNOS was also diminished by the TK-CuNPs. TK-CuNPs did not possessed toxicity to the RAW-264.7 cells and suppressed the inflammatory markers level in the RAW-264.7 cells. Discussion and Conclusion: The current findings revealed that fabricated TK-CuNPs can be a hopeful therapeutic agent to treat acute colitis.

Copyright of Chinese Journal of Applied Ecology / Yingyong Shengtai Xuebao is the property of Chinese Journal of Applied Ecology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Industrial development, along with the rapid growth of the economy, has greatly improved the quality of life in humans. Moreover, advancements in medical technology have increased life expectancy. Small particles increase airway inflammation when they penetrate the alveoli. We observed that GHX02 decreased the frequency and delayed the onset time of citric acid-induced coughing in guinea pigs. A phenol red secretion assay indicated that the GHX02 extract exhibits potent expectorant activity. The GHX02 extract also greatly reduced leukocyte levels. Our results indicate that GHX02 inhibits airway inflammation, reduces sputum production, and relieves cough. The GHX02 extract suppressed histamine release from mast cells resulting from compound 48/80-induced degranulation. The extract exhibited antimicrobial activity against Streptococcus pneumoniae and significantly inhibited the formation of LTC4. At high concentrations, the GHX02 extract suppressed the formation of PGE2 (prostaglandin E2). Interleukin (IL)-4 and IL-13 levels decreased with an increasing dosage of GHX02. Oral administration of the GHX02 extract suppressed PM10D-induced inflammatory symptoms in the lung, including increased alveolar wall thickness, accumulation of collagen fibers, and cytokine release. Treatment with the GHX02 extract also resulted in lower levels of inflammatory cells, in bronchoalveolar lavage fluid and lung tissue. Our results indicate that GHX02 may be a useful therapeutic agent for treatment of respiratory diseases. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The genome sequence of a novel RNA virus, Trichosanthes associated rhabdovirus 1 (TrARV1), was identified in a transcriptome dataset isolated from a root sample of Trichosanthes kirilowii, which is a flowering plant belonging to the family Cucurbitaceae. The fruits, seeds, and root tubers of T. kirilowii have been used clinically in traditional Chinese medicine. The TrARV1 genome sequence was predicted to have six open reading frames (ORFs) encoding fi ve canonical structural proteins of the family Rhabdoviridae (N ORF for nucleocapsid, P ORF for phosphoprotein, M ORF for matrix protein, G ORF for glycoprotein, and L ORF for polymerase), and an accessory protein. Sequence comparisons and phylogenetic analyses based on L and N proteins confi rmed that TrARV1 is a novel member of the genus Cytorhabdovirus of the family Rhabdoviridae. TrARV1 is most closely related to Wuhan insect virus 5 and persimmon virus A. The putative cis-regulatory elements involved in transcription termination and polyadenylation, commonly found in the gene junction regions of rhabdoviruses, were also identifi ed in the TrARV1 genome having the consensus sequence 3'- ACUAAAUUAUUUUGAUCUUU-5'. The genome sequence of TrARV1 may be useful to study the evolution and molecular biology of cytorhabdoviruses. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii Maxim. is a vital traditional herbal medicinal plant found in northeastern Asia. Its roots, fruits, and seeds are used as food and medicine. Roots harvested for medicinal use take over 3 yr to mature when the plant is grown in a traditional way through cultivation in the field. This coupled with uncertainty in identification of the plant when collected from the wild calls for a standard in vitro propagation system to meet the increasing demand for it. The purpose of this study was to develop a standard protocol for the in vitro micropropagation of T. kirilowii. Ten different media supplemented with different concentrations of plant growth regulators were evaluated. At 5 wk, De Greef and Jacobs medium supplemented with 0.1 mg L−1 kinetin led to optimal shoot growth, while the same medium supplemented with 0.5 mg L−1 indole 3-butyric acid induced optimal root growth, also at 5 wk. The micropropagated plants that were acclimatized for 8 wk in the greenhouse produced mature root tubers after planted in the field for 3 mo. Therefore, these findings provide a basis for future large-scale in vitro propagation of T. kirilowii. [ABSTRACT FROM AUTHOR]

Most herbs of traditional Chinese medicine (TCM) are used as air-dried decoction pieces that are manufactured and kept at ambient temperature for long periods. Given the ability of some desiccation-tolerant plants to conserve RNA, it could be worthwhile to isolate mRNA from TCM decoction pieces as part of a transcriptomic strategy to identify new substances with potential pharmaceutical application. Here, we report the molecular cloning of a novel trypsin inhibitor (as the probable alleleic variants TKTI-2 and TKTI-3) from the decoction piece of Radix Trichosanthis, representing the dried root of Trichosanthes kirilowii. From this material, the total RNA was extracted and a cDNA library was constructed from the isolated mRNA from which the cDNAs of two precursors were successfully cloned and sequenced. TKTI-3 showed an amino-acid substitution in the otherwise highly-conserved P1-P1′ reaction site of the mature peptide, which we confirmed to not be an artefact. Subsequent analysis using LC-MS/MS detected the presence of specific tryptic peptides expected from TKTI-3, confirming the presence and expression of this locus in Radix Trichosanthis. More generally, this study indicates that mRNA can persist in decoction pieces and so could present a viable option for the molecular cloning from other TCMs. • Success of isolation of RNA using decoction pieces of Chinese medicine, Radix Trichosanthis. • The probable allelic variants of a novel trypsin inhibitor were successfully cloned and sequenced using the cDNA library. • Parallel analysis using LC-MS/MS to determine the amino acid mutation of trypsin inhibitor peptide. [ABSTRACT FROM AUTHOR]

Highlights • A hierarchical extraction method was established for determination flavonoids and triterpenes in T. kirilowii. • An UHPLC-MS/MS method was established for quantification multiple compounds. • Reveal the distribution of resource components in different parts of T. kirilowii. • The results could be useful for the rational utilization of T. kirilowii. Abstract Trichosanthin (root of Trichosanthes kirilowii Maxim.) has received much attention, but T. kirilowii has received little systematic analysis. A comprehensive evaluation of flavones and triterpenes in nine different parts of T. kirilowii would, thus, be of value to fully understand the potential medicinal properties of T. kirilowii. A hierarchical extraction—simultaneous determination method was established, which based on different types of components had different response on the MS detector. The hierarchical extraction method extracted 3, 29-dibenzoyl rarounitriol solely, which response lowly on the MS detector in the mixture of one step extract, in order to obtain accurate results in the next UHPLC-MS/MS assay. By this method, ten flavones, three tetracyclic triterpenoids, and one pentacyclic triterpenoid in the extracts of T. kirilowii were determined. The results showed the level of flavonoids in leaves was higher than those in the other samples, tetracyclic triterpenoids mainly existed in root and root bark, and pentacyclic triterpenoid, especially 3, 29-dibenzoyl rarounitriol was concentrated in the seed kernel. According to principal component analysis, the F values of the leaves of all varieties were generally greater than those in other parts. The leaves might provide a supplementary source of flavonoids, and the root and root bark could be a good choice for tetracyclic triterpenoids. The analysis results revealed the distribution of flavones and triterpenes in different parts of T. kirilowii and provided reference for the research and rational utilization of T. kirilowii. [ABSTRACT FROM AUTHOR]

Carboxymethyl chitosan and resistant starch exhibit good performance in diabetes regulation. We prepared carboxymethyl chitosan - resistant starch complex. Test the properties of composite resistant starch by using X-ray diffraction, water contact angle, infrared spectroscopy, and scanning electron microscopy, interactions with intestinal microbiota and mouse experiments were also conducted. The results indicated that the composite resistant starch had a good effect on promoting the proliferation of probiotics on Bifidobacterium and a significant inhibitory effect on Escherichia coli than resistant starch (P < 0.05). After administration, the water intake and weight of diabetic mice were significantly reduced. The blood glucose of diabetic mice was also reduced, and oral glucose tolerance showed that the glucose degradation rates of composite resistant starch were significantly improved compared to model mice. Cholesterol, triglycerides, high-density lipoprotein and low-density lipoprotein were significantly lower than those in the diabetes group (P < 0.05). The diversity of the gut microbiota was also proven. • Carboxymethyl chitosan can enhance the efficacy of resistant starch. • Carboxymethyl chitosan-resistant starch complex can adjust the gut flora better the resistant starch. • Carboxymethyl chitosan-resistant starch complex can improve the diabetes mice better the resistant starch. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Two new sterols [trichosanhemiketal A, B; (1 – 2)] and 13 known compounds were isolated from Trichosanthes kirilowii root. • The 13,14- seco -13,14-epoxyporiferastane skeleton (1 – 2) was found from the Cucurbitaceae family for the first time. • Cucurbitacins 4 , 5 , 6 , and 8 showed significant NO inhibitory activities. • Compound 4 inhibited inducible iNOS and COX-2 expression. Abstract Of the 32 Trichosanthes species in China, T. kirilowii Maxim. is the most renowned species used in traditional Chinese medicine and has diverse pharmacological properties. However, most of the phytochemical studies of T. kirilowii have focused on the fruits and seeds. In our investigation of the chemical constituents of T. kirilowii roots, two previously undescribed sterols [trichosanhemiketal A and B (1 and 2)], together with 13 known compounds, were isolated and their structures were elucidated. To the best of our knowledge, this represents the first isolation of compounds with a 13,14- seco -13,14-epoxyporiferastane (1 – 2) skeleton from the Cucurbitaceae family. The anti-inflammatory activity of the isolated compounds was determined through an analysis of their inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in macrophage RAW264.7 cells. Of the compounds, 4 , 5 , 6 , and 8 showed significant inhibitory activities, with IC 50 values of 8.5, 15.1, 25.4, and 28.5 µM, respectively. In addition, compound 4 inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expression in a concentration-dependent manner. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii Maxim. is a climbing herb with considerable medicinal value. In this study, efficient protocols for callus-mediated regeneration and in vitro tuberization of this plant were developed. Sterilized stem and leaf tissues were cultured on Murashige and Skoog (MS) medium with plant growth regulators (PGRs), and additives that promoted callus induction and regeneration. Both stem and leaf tissues showed the best response (100%) for callus initiation on MS medium supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D). Efficient shoot organogenesis was obtained by exposing the callus tissue to 4.6-μM kinetin, 2.2-μM 6-benzylaminopurine, and 2.7-μM 1-naphthylacetic acid (NAA) along with 12.6-μM copper sulfate, which yielded a shoot regeneration rate of 85.5% and 28 shoots derived from each callus. In vitro shoots were best rooted on half-strength (1/2) MS medium with 2.7-μM NAA. Tuberous roots were efficiently induced on rooting medium with 5% (w/v) sucrose under short illumination conditions (8 h photoperiod). Rooted plantlets were successfully acclimatized in pots with a > 90% survival rate. This protocol provides an effective method for callus-mediated regeneration and in vitro root tuberization. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ten new terpenoids were isolated from the pericarps of Trichosanthes kirilowii. • Seventeen terpenoids (1 – 15 , 18 , and 20) exhibited anticomplement activity. • The long aliphatic chains might enhance the monoterpenoids' anticomplement activity. • Anticomplement terpenoids 8 and 11 obviously attenuated H1N1-induced ALI in vivo. The pericarps of Trichosanthes kirilowii are often used to treat cough in traditional Chinese medicine, and its ethanol extract exhibited effective therapeutic effects on acute lung injury (ALI) in vivo caused by H1N1. An anticomplement activity-guided fractionation on the extract resulted in the isolation of ten new terpenoids, including seven monoterpenoids, trichosanates A–G (1 – 7), and three cucurbitane-type triterpenoids, cucurbitacins W–Y (8 – 10), as well as eleven known terpenoids (11 – 21). The new terpenoids' structures were determined by spectroscopic analysis, X-ray crystallographic analysis (1), electronic circular dichroism (ECD) analysis and calculations (2 – 10). Twelve monoterpenoids (1 – 7 and 11 – 15) and five cucurbitane-type triterpenoids (8 – 10 , 18 , and 20) exhibited anticomplement activity in vitro. For the monoterpenoids, the long aliphatic chain substituents might enhance their anticomplement activity. Additionally, two representative anticomplement terpenoids, 8 and 11 , obviously attenuated H1N1-induced ALI in vivo by inhibiting complement overactivation and reducing inflammatory responses. [ABSTRACT FROM AUTHOR]

A 46-year-old Korean woman was diagnosed with stage III breast cancer and underwent 8 cycles of neoadjuvant chemotherapy, breast conservation surgery and adjuvant radiotherapy. However, the cancer recurred in the right upper lung (RUL) and the right pulmonary hilum after 8 months. The RUL nodule was removed through a wedge resection, and the pathologic finding was revealed as a metastatic adenocarcinoma. Adjuvant chemotherapy was recommended, but she refused it because she feared adverse reactions to chemotherapy. Instead, Korean Medicine Therapy with intravenous wild ginseng pharmacopuncture (WGP), Cordyceps sinensis pharmacopuncture, Trichosanthes kirilowii pharmacopuncture, Euonymus alatus pharmacopuncture (EAP) and Astragalus membranaceus pharmacopuncture was started. After a month, the disease looked stable, but findings of newly occurring metastatic lymphadenopathies appeared on CT after 6 months. Salvage chemotherapy was recommended, but she also refused it. At this time, Prunella vulgaris pharmacopuncture was started. Finally, a complete resolution was confirmed on PET-CT after 5 months, and she has remained in stable condition for more than 6 months with WGP, EAP, a Soram nebulizer solution inhalation and the oral intake of Soramdan S and Hangamdan S.

Ultraviolet B (UVB) radiation induces DNA damage, oxidative stress and inflammation, and suppresses the immune system in the skin, which collectively contribute to skin aging and carcinogenesis. The DNA damage response, including DNA repair, can be regulated by the circadian clock and microRNA (miRNA) expression. The aim of the present study was to evaluate the reparative action of Trichosanthes kirilowii extract (TKE) against UVB irradiation-induced DNA damage in human keratinocytes. TKE demonstrated low cytotoxicity in normal HaCaT keratinocytes at low doses (up to 100 µg/ml). The results of a comet assay revealed that TKE enhanced the repair of UVB-induced DNA damage. TKE significantly upregulated the expression of the core clock protein, brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein-1 (BMAL1), and downregulated the expression of miRNA (miR)-142-3p, as demonstrated using western blotting and the reverse transcription-quantitative polymerase chain reaction. Furthermore, the suppression of miR-142-3p by a specific inhibitor positively correlated with the repair activity. Overall, the data obtained demonstrated that TKE enhanced the repair of UVB-induced DNA damage by regulating the expression of BMAL1 and miR-142-3p. Consequently, TKE can be considered a potential candidate for the treatment of skin diseases associated with UVB-induced damage. [ABSTRACT FROM AUTHOR]

Punicic acid (PuA) is a conjugated linolenic acid (C18:3Δ) with a wide range of nutraceutic effects with the potential to reduce the incidence of a number of health disorders including diabetes, obesity, and cancer. It is the main component of seed oil from Punica granatum and Trichosanthes kirilowii. Previously, production of relatively high levels of this unusual fatty acid in the seed oil of transgenic Arabidopsis thaliana plant was accomplished by the use of A. thaliana fad3/fae1 mutant high in linoleic acid (18:2∆) and by co-expression of P. granatum FATTY ACID CONJUGASE ( PgFADX) with Δ12-DESATURASE ( FAD2). In the current study, P. granatum cDNAs governing PuA production were introduced into the yeast Schizosaccharomyces pombe. Expression of PgFADX alone resulted in production of PuA at the level of 19.6% of total fatty acids. Co-expression PgFADX with PgFAD2, however, further enhanced PuA content to 25.1% of total fatty acids, the highest level reported to date for heterologous expression. Therefore, microbial systems can be considered as a potential alternative to plant sources for a source of PuA for nutraceutic applications. [ABSTRACT FROM AUTHOR]

An efficient and green protocol for the synthesis of isoaurones by condensation of benzofuranone with benzaldehydes using polyethylene glycol (PEG) is demonstrated. Excellent yields, no reagents/additional catalysts,E-selectivity, short reaction times, and no work-up/extraction makes an attractive benign procedure. Additionally, 4′,6-dihydroxy-4-methoxyisoaurone isolated fromTrichosanthes kirilowiihas been synthesized for the first time and confirmed its geometry asE. [ABSTRACT FROM AUTHOR]

In conducting this study, the extraction of Trichosanthes kirilowii maxim seed oils (TSO) was carried out with the help of cold pressing (CP), hot pressing (HP) and soxhlet extraction (SE). Investigation, together with comparison, was carried out with respect to the physicochemical properties, thermal stability and antioxidant action of TSO. The key ingredients in the seeds consisted of fat, fiber and protein. The physicochemical characteristics of the oils brought to light the fact that CPTSO possessed top oil quality. The findings also suggested that linoleic acid, punicic acid and oleic acid were the leading unsaturated fatty acids in TSO. It was also discovered that TSO had an almost identical chemical composition regardless of the extraction method was used. It was demonstrated by TG/ DTG curves that both HPTSO and CPTSO had more thermal stability in comparison with SETSO. Furthermore, the antioxidant activity assessments emphasized that CPTSO had better radical scavenging potential. CP had the ability to deliver an extract with higher quality as well as antioxidant activity in comparison with HP and SE methods and can be taken into consideration as a more suitable method in order to attain high quality oil. [ABSTRACT FROM AUTHOR]

This report presents a case that shows a significant anticancer effect of Korean medicine therapy (KMT). A 79-year-old man, who was diagnosed as stage Ⅳ non-small cell lung cancer (NSCLC) in December 2012, was treated with KMT including intravenous pharmacopunctures and oral herbal medicine from February 22, 2013, until September 2013 without any surgical intervention, chemotherapy or radiotherapy. The intravenous pharmacopunctures were the wild ginseng pharmacopuncture, Cordyceps sinensis pharmacopuncture and Trichosanthes kirilowii pharmacopuncture. The oral herbal medicine used was soramdan, made of cultivated wild ginseng. The effectiveness of this therapy was evaluated with computed tomography and the Eastern Cooperative Oncology Group (ECOG) performance scale. The size of the tumor mass was markedly decreased and the ECOG performance scale was also improved. These results suggest that KMT alone can be an effective method to treat NSCLC.

Background: Diabetes is a serious chronic metabolic disorder. Trichosanthes kirilowii Maxim. (TK) is traditionally used for the treatment of diabetes in traditional Chinese medicine (TCM). However, the clinical application of TK on diabetic patients and the hypoglycemic efficacies of TK are still unclear. Methods: A retrospective cohort study was conducted to analyze the usage of Chinese herbs in patients with type 2 diabetes in Taiwan. Glucose tolerance test was performed to analyze the hypoglycemic effect of TK. Proteomic approach was performed to identify the protein constituents of TK. Insulin receptor (IR) kinase activity assay and glucose tolerance tests in diabetic mice were further used to elucidate the hypoglycemic mechanisms and efficacies of TK. Results: By a retrospective cohort study, we found that TK was the most frequently used Chinese medicinal herb in type 2 diabetic patients in Taiwan. Oral administration of aqueous extract of TK displayed hypoglycemic effects in a dose-dependent manner in mice. An abundant novel TK protein (TKP) was further identified by proteomic approach. TKP interacted with IR by docking analysis and activated the kinase activity of IR. In addition, TKP enhanced the clearance of glucose in diabetic mice in a dose-dependent manner. Conclusions: In conclusion, this study applied a bed-to-bench approach to elucidate the hypoglycemic efficacies and mechanisms of TK on clinical usage. In addition, we newly identified a hypoglycemic protein TKP from TK. Our findings might provide a reasonable explanation of TK on the treatment of diabetes in TCM. [ABSTRACT FROM AUTHOR]

Background: The androgen comprises a group of hormones that play roles in male reproductive activity as well as personal characteristics. Objective: We investigated the androgenic activity of various herbal medicines in human prostate cancer 22Rv1 cells. Materials and Methods: Herbal extracts of Trichosanthes kirilowii (TK), Asarum sieboldii (AS), Sanguisorba officinalis (SO), and Xanthium strumarium (XS) were selected to have androgenic effects based on a preliminary in vitro screening system. Results: TK, AS, SO, and XS enhanced the proliferation of 22Rv1 cells without having cytotoxic effects. All tested herbal extracts increased androgen receptor (AR)-induced transcriptional activity in the absence or presence of dihydrotestosterone (DHT). In an AR-binding assay, TK, but not AS, SO, or XS, produced a significant inhibition of AR binding activity, indicating it has androgenic activity. Additionally, TK treatment positively regulated mRNA expression of the AR-related molecular targets prostate-specific antigen (PSA) and kallikrein 2 (KLK2) compared with untreated control. Conclusion: Taken together, TK-enhanced AR-mediated transcriptional activity might be an attractive candidate drug for treating androgen-related diseases. [ABSTRACT FROM AUTHOR]

The physicochemical properties and volatile profiles of cold-pressedTrichosanthes kirilowiiMaxim (T. kirilowii) seed oils from four regions in China were determined in this study. The total oil content and cold-pressed oil yield of the four different sourced seeds were 38.06–44.33% and 15.17–30.97%, respectively. All the cold-pressed oil samples were found to be rich in polyunsaturated fatty acids, with content ranging from 45.41 to 75.32% of the total fatty acids. Punicic, α-eleostearic and catalpic acids were the main conjugated linolenic acid isomers in the cold-pressedT. kirilowiiseed oils. The results of melting and crystallization profiles indicated that each oil sample exhibited different transitions steps due to its triacylglycerol composition, crystal structure and total unsaturation. Analysis of volatile profiles showed that 2,4-nonadienal was one of the most important aldehydes in the cold-pressedT. kirilowiiseed oils, and less short chain acids (0.20%) but more esters (5.48%) were found in the sample with high content of punicic acid (Hebei sample). Results of oil quality indices indicated that cold-pressedT. kirilowiiseed oils were liable to be oxidized, and their stabilities reduced with the increase of acid values. In general, more attention should be paid to improve the oxidative stability of cold-pressedT. kirilowiiseed oils in their further application in food industry. [ABSTRACT FROM AUTHOR]

Tomato mottled mosaic virus (ToMMV) was first identified in tomato in Mexico (Li et al. 2013). It belongs to the genus Tobamovirus and family Virgaviridae, and is a positive-sense single-stranded RNA virus. The viral genome contains about 6400 nucleotides, encoding four proteins, including the 126 K protein, 183 K protein, movement protein (MP) and coat protein (CP) (Tu et al. 2021). ToMMV mainly poses a serious risk to solanaceous crops. The virus-infected plants appear stunted growth and top necrosis, and the disease leaves show mottled, shrinkage and necrosis symptoms, resulting in a significant decline in tomato fruit yield and quality (Li et al. 2017; Tu et al. 2021). Chinese snake gourd (Trichosanthes kirilowii Maxim) is a perennial climbing herb in the family Cucurbitaceae, and the fruit, seed, peel and root can all be used as traditional Chinese medicine. In May of 2021, twenty-seven symptomless seedlings (developed from tissue culture plantlets) were randomly collected from nursery in Fengyang, Anhui Province. Total RNA of each sample was extracted, and RT-PCR was performed using degenerate tobamovirus primers Tob-Uni1 (5'-ATTTAAGTGGASGGAAAAVCACT-3') and Tob-Uni2 (5'-GTYGTT GATGAGTTCRTGGA-3') (Letschert et al. 2002). Amplicons with expected size were obtained from 6 of 27 samples and sequenced. Alignment results showed that the nucleotide sequence identities ranged from 98.7 to 100% with all ToMMV isolates deposited in NCBI GenBank. Then, ToMMV coat protein (CP) gene was amplified using specific primers CP-F (5'-ATGTCTTACGCTATTACTT CTCCG-3') and CP-R (5'-TTAGGACGCTGGCGCAGAAG-3'). The CP fragment was obtained and sequenced. Sequence alignment indicated that CP sequence of isolate FY (GenBank accession no. ON924176) exhibited a 100% identity with ToMMV isolate LN (MN853592.1). The anti-ToMMV polyclonal antibody (PAb) was prepared by the author (S.L.) by immunizing rabbit with purified virus from Nicotiana benthamiana, and serological tests (dot-enzyme linked immunosorbent assay, Dot-ELISA) of RNA-positive T. kirilowii leaf samples using anti-ToMMV PAb were also positive. To fulfill a Koch's postulate, a pure culture of ToMMV was obtained from N. benthamiana using infectious cDNA clone of ToMMV (Tu et al. 2021), and then healthy T. kirilowii plants were mechanically inoculated with a prepared inoculum from ToMMV-infected N. benthamiana, as described previously (Sui et al. 2017). T. kirilowii seedlings showed chlorosis and leaf tip necrosis symptoms at 10 and 20 day post-inoculation respectively, and ToMMV infection on symptomatic plants was also verified by RT-PCR detection using primers CP-F and CP-R. These results demonstrated that T. kirilowii is a host of ToMMV under natural conditions, which might threaten the production of this medicinal plant. The seedlings from nursery appeared to be asymptomatic, but the plants showed chlorosis and necrosis symptoms after indoor inoculation. In qRT-PCR analysis, viral accumulation level in greenhouse-inoculated plants was a 25.6-fold of that in field-collected samples, which may be the reason of different symptom expression between field samples and inoculated samples. ToMMV has now been detected from the solanaceous (tomato, pepper and eggplant) and leguminous (pea) crops in the field (Li et al. 2014; Ambrós et al. 2017; Zhang et al. 2022). To our knowledge, this is the first report of natural infection of ToMMV in T. kirilowii as well as its natural infection on Cucurbitaceae plants.

A rapid, sensitive and high-throughput liquid chromatography-tandem mass spectrometry was established and validated to assay the concentrations of 3,29-dibenzoyl rarounitriol in rat plasma. Plasma samples were processed by liquid-liquid extraction with ethyl acetate and separated on a Hypersil Gold C18 column (50 × 4.6 mm, 3 µm) at an isocratic flow rate of 0.5 mL/min using methanol-10 m m ammonium acetate-formic acid (90:10:0.1, v/v/v) as mobile phase. The total run time was 5 min for each sample. MS/MS detection was accomplished in selected reaction monitoring mode with positive electrospray ionization. The calibration curve was linear over the concentration range of 0.125-50 ng/mL with lower limit of quantification of 0.125 ng/mL. The intra- and inter-day precisions were <10.1% in terms of coefficient of variation, and the accuracy was within ±11.7% in terms of relative error. The developed method was successfully applied to a pharmacokinetic study of 3,29-dibenzoyl rarounitriol following intragastric administration of 3.65 mg/kg to Wistar rats. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

One new lignan, trichobenzolignan ( 1), and seven known compounds, ligballinol ( 2), (−)-pinoresinol ( 3), ehletianol C ( 4), luteolin 7- O-β-D-glucopyranoside ( 5), chrysoeriol-7- O-β-D-glucopyranoside ( 6), 10α-cucurbita-5,24-dien-3β-ol ( 7), and arvenin I ( 8). Their structures were established on the basis of spectral and chemical evidence, which were in agreement with those reported in literature. The cytotoxic activities of these compounds were evaluated on four cancer cell lines such as A-549 (human lung cancer), HT-29 (human colon adenocarcinoma), OVCAR (human ovarian carcinoma), and MCF-7 (human breast cancer). As the results, compound 7 showed significant activity on HT-29 and OVCAR cancer cell lines with IC of 4.1 and 6.5 µM, respectively. Compounds 1, 5, 6, and 8 exhibited moderate activities in all cancer cell lines with IC ranging from 11.3 to 42.8 µM. [ABSTRACT FROM AUTHOR]

Root extract of Boerhaavia diffusa L. induced systemic resistance in tobacco against Tobacco mosaic virus. A 30 kDa protein was isolated as the active component, called BDP-30 on the basis of the molecular weight and source plant. BDP-30, a glycoprotein, was found to be temperature and protease resistant. It was basic, possessing a pI greater than 9.0. In-gel proteolytic digestion of BDP-30 generated two peptides that possessed the amino acid sequence KLYDIPPLR and KVTLPYSGNYER by LC/MS/MS. Both peptides shared absolute sequence identity with trichosanthin, a ribosome-inactivating protein from Trichosanthes kirilowii, and a 78% and 100% homology respectively with an RIP from Bryonia dioica, bryodin. Further, effort was made to look at the fate of TMV in induced resistant Nicotiana tabacum cv. Xanthi, a systemic host of the virus, at specified days after inoculation in control and treated plants. TMV coat protein (CP) was detected by immunoblot 7 days post inoculation up to 21 days in the control set, but not in treated resistant plants. TMV RNA was detected by RT-PCR using TMV-CP specific primers. Resistant tobacco did not show presence of TMV RNA up to 21 days of inoculation. This suggests that BDP-30 may be suppressing TMV replication. [ABSTRACT FROM AUTHOR]

Anthracnose disease is an important fungal disease on Trichosanthes kirilowii Maxim. ( T. kirilowii), which has recently become a major problem in production areas of T. kirilowii in Anhui Province, central China. A total of 78 putative Colletotrichum spp. isolates were obtained from infected leaves and fruits of T. kirilowii showing anthracnose symptoms during the years 2009-12. All the fungal isolates were consistently identified as Colletotrichum gloeosporioides by a combination of morphological characteristics, species-specific PCR assays and sequence analysis of the ITS region. The results of pathogenicity tests showed difference in virulence among 24 isolates of C. gloeosporioides representatively selected from different regions, when disease symptoms were evaluated after artificially inoculation on the detached T. kirilowii fruits for 2 weeks. The mycelial growth and germination rate of C. gloeosporioides isolates in vitro were determined under temperatures of 10, 15, 20, 25, 28, 30, 33 and 36°C. The optimum growth temperatures of these isolates were observed at 25-30°C, and the germination rates were rather high in the range of 20-33°C. Host range study revealed that the pathogen, under experimental conditions, can infect nine cultivated and weed species belonging to six families (Cucurbitaceae, Rosaceae, Solanaceae, Convolvulaceae, Celastraceae and Cannabinaceae). Moreover, C. gloeosporioides can overwinter safely in infected tissues in field soil, suggesting the potential contribution of plant debris to disease epidemics in the region. The study presents useful information for epidemiological surveillance of T. kirilowii anthracnose caused by C. gloeosporioides in the central region of China. [ABSTRACT FROM AUTHOR]

Context: Three-phase partitioning (TPP), a unique technique which has been explored for protein separation, was used for extraction of trichosanthin (TCS). Objective: TPP was used to optimize the TCS extraction and to determine its anticancer activity. Materials and methods: The process consists of the simultaneous addition of t-butanol and ammonium sulfate to the aqueous slurry of Trichosanthes kirilowii Maxim (Cucurbitaceae) root powder. The extraction of TCS was optimized with respect to the concentration of ammonium sulfate loading, the ratio of t-butanol to slurry, extraction time and pH. The anticancer activity was performed using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay in vitro. Results: The extraction time with this technique is lower in comparison to conventional solvent extraction. The optimized protocol resulted in maximum recovery of 98.68% (w/w) protein within 1 h. The in vitro cytotoxic activity of the TCS was evaluated against HepG2 and WRL 68 cancer cell line and results showed that TCS possesses quite highly significant anticancer activity having IC50 values of 10.38 and 15.45 μmol/l, respectively, comparable to standard drugs. Conclusion: This framework is utilized as a basis for optimization for protein separation using TPP technique which is economical and eco-friendly. [ABSTRACT FROM AUTHOR]