An Injectable Puerarin Depot Can Potentiate Chimeric Antigen Receptor Natural Killer Cell Immunotherapy Against Targeted Solid Tumors by Reversing Tumor Immunosuppression.

Chimeric antigen receptor natural killer (CAR-NK) cell therapy represents a potent approach to suppressing tumor growth because it has simultaneously inherited the specificity of CAR and the intrinsic generality of NK cells in recognizing cancer cells. However, its therapeutic potency against solid tumors is still restricted by insufficient tumor infiltration, immunosuppressive tumor microenvironments, and many other biological barriers. Motivated by the high potency of puerarin, a traditional Chinese medicine extract, in dilating tumor blood vessels, an injectable puerarin depot based on a hydrogen peroxide-responsive hydrogel comprising poly(ethylene glycol) dimethacrylate and ferrous chloride is concisely developed. Upon intratumoral fixation, the as-prepared puerarin depot (abbreviated as puerarin@PEGel) can activate nitrogen oxide production inside endothelial cells and thus dilate tumor blood vessels to relieve tumor hypoxia and reverse tumor immunosuppression. Such treatment can thus promote tumor infiltration, survival, and effector functions of customized epidermal growth factor receptor (HER1)-targeted HER1-CAR-NK cells after intravenous administration. Consequently, such puerarin@PEGel-assisted HER1-CAR-NK cell treatment exhibits superior tumor suppression efficacy toward both HER1-overexpressing MDA-MB-468 and NCI-H23 human tumor xenografts in mice without inducing obvious side effects. This study highlights a potent strategy to activate CAR-NK cells for augmented treatment of targeted solid tumors through reprogramming tumor immunosuppression.

Dietary polyunsaturated fatty acids intake, air pollution, and the risk of lung cancer: A prospective study in UK biobank.

BACKGROUND: Epidemiological evidence on the association between specific types of polyunsaturated fatty acids (PUFAs) intake and lung cancer risk is limited. However, whether dietary-specific PUFAs intake can modify the association between air pollutants and incident lung cancer remains unknown. METHODS: Cox proportional hazard models and restricted cubic spline regression were used to evaluate the associations of omega-3 PUFAs, omega-6 PUFAs and the ratio of omega-6 PUFAs to omega-3 PUFAs intake with lung cancer risk. Furthermore, we evaluated the associations between air pollutants and incident lung cancer, and whether dietary-specific PUFAs intake would modify the relationship using stratification analyses. RESULTS: This study found significant associations between the risk of lung cancer and omega-3 PUFAs intake (hazard ratio [HR], 0.82; 95 % confidence interval [CI], 0.73-0.93; per 1 g/d), and omega-6 PUFAs intake (HR, 0.98; 95 % CI, 0.96-0.99; per 1 g/d). We did not observe an association between the omega-6 to omega-3 PUFAs intake ratio and incident lung cancer. With regard to air pollution, omega-3 PUFAs intake attenuated the positive relationship between nitrogen oxides (NOx) pollution and lung cancer risk, and an increased incidence of lung cancer was found only in the low omega-3 PUFAs intake group (p < 0.05). Surprisingly, PUFAs intake (regardless of omega-3 PUFAs, omega-6 PUFAs, or in total) reinforced the pro-carcinogenic effects of PM2.5 on lung cancer, and a positive association between PM2.5 pollutants and incident lung cancer was observed only in the high PUFAs groups (p < 0.05). CONCLUSIONS: Higher dietary omega-3 and omega-6 PUFAs intake was associated with a decreased risk of lung cancer in the study population. As omega-3 PUFAs have different modification effects on NOX and PM2.5 air pollution related lung cancer incidence, precautions should be taken when using omega-3 PUFAs as health-promoting dietary supplements, especially in high PM2.5 burden regions.

Understanding the Origin and Evolution of Tea (Camellia sinensis [L.]): Genomic Advances in Tea.

Tea, which is processed by the tender shoots or leaves of tea plant (Camellia sinensis), is one of the most popular nonalcoholic beverages in the world and has numerous health benefits for humans. Along with new progress in biotechnologies, the refined chromosome-scale reference tea genomes have been achieved, which facilitates great promise for the understanding of fundamental genomic architecture and evolution of the tea plants. Here, we summarize recent achievements in genome sequencing in tea plants and review the new progress in origin and evolution of tea plants by population sequencing analysis. Understanding the genomic characterization of tea plants is import to improve tea quality and accelerate breeding in tea plants.

Selenium treatment promotes anthocyanin accumulation in radish sprouts (Raphanus sativus L.) by its regulation of photosynthesis and sucrose transport.

Red radish sprout become a popular dietary vegetable because of its unique flavor, abundant nutrients and short production cycle. As a cruciferous plant, it has strong ability to absorb and assimilate Se which can promote the content of anthocyanin in plants. However, the mechanisms of Se on anthocyanin accumulation are still unclear. In this study, we explored that appropriate Se promoted growth, antioxidant system and nutrients in radish sprouts. The enhancement of photosynthesis by Se treatment resulted in more sucrose synthesis in radish sprouts. And the transport of sucrose from cotyledon to hypocotyl promoted by Se through up-regulating the gene expression of sucrose transporters, and more sucrose increased the expression of anthocyanin biosynthesis genes to promote anthocyanin accumulation in hypocotyl. These results reveal the beneficial effect of Se on radish sprouts quality, and provide a new insight into the function of Se on sucrose-induced anthocyanin accumulation in radish sprouts.

Peptide signaling in anther development and pollen-stigma interactions.

Polypeptides play irreplaceable roles in cell-cell communication by binding to receptor-like kinases. Various types of peptide-receptor-like kinase-mediated signaling have been identified in anther development and male-female interactions in flowering plants. Here, we provide a comprehensive summary of the biological functions and signaling pathways of peptides and receptors involved in anther development, self-incompatibility, pollen tube growth and pollen tube guidance.

The lineage-specific evolution of the oleosin family in Theaceae.

Oleosins play essential roles in stabilization of lipid droplets (LDs) and seed oil production. However, evolution of this gene family has not been reported in Theaceae, a large plant family that contains many important tea and oil tea species. In this study, a total of 65 oleosin genes were identified in nine genome-sequenced Theaceae species. Among these genomes, the gene number of oleosin showed significant difference, with Camellia sinensis var. sinensis cv. Shuchazao and Camellia lanceoleosa displayed more oleosin numbers than other species. Phylogenetic analyses revealed that Theaceae oleosin genes were classified into three clades (U, SL, SH) respectively. Proteins within the same clade had similar gene structure and motif composition. Segmental duplication was the primary driving force for the evolution of oleosin genes in Shuchazao (SCZ), Huangdan (HD), C.lanceoleosa (Cla), and wild tea (DASZ). Synteny analysis showed that most oleosin genes displayed inter-species synteny among tea and oil tea species. Expression analysis demonstrated that oleosin genes were specifically expressed in seed and kernel of Huangdan (HD) and C.lanceoleosa. Moreover, expression divergence was observed in paralogous pairs and ∼1-2 oleosin genes in each clade have become activate. This study leads to a comprehensive understanding of evolution of oleosin family in Theaceae, and provides a rich resource to further address the functions of oleosin in tea and oil tea species.

Qiangxin recipe improves doxorubicin-induced chronic heart failure by enhancing KLF5-mediated glucose metabolism.

BACKGROUND: Qiangxin recipe (QXF) is a well-known Chinese herbal medicine commonly used in Asia for thousands of years to treat cardiovascular diseases, but its underlying mechanism remains unclear. PURPOSE: This study aimed to illustrate whether Qiangxin Recipe (QXF) induce glucose metabolism and inhibit cardiomyocyte apoptosis by promoting the activation of the transcription factor Krüppel like factor 5 (KLF5). MATERIAL AND METHODS: In vitro experiments, we constructed an H9C2 cardiomyocyte injury model using doxorubicin and used RNA-seq data analysis to detect the mechanism of QXF. In in vivo experiments, C57 BL/6 mice injected with doxorubicin (4 mg/kg every 6 days, for 30 days) to construct a CHF mouse model and randomly divided into to the normal control group, Dox group and Dox+QXF group (2.12 g/kg/day, 4.24 g/kg/day, for 30 days). Using Echocardiography, serum biochemical indices BNP, cTnl; and histopathological tests involving HE staining, Tunel staining and Immuno-dual fluorescence colocalization to analyze the therapeutic mechanism of QXF. RESULTS: We verified that the Qiangxin recipe could reverse cardiomyocyte dying through enhancing glucose metabolism and reducing apoptosis to improve CHF. Mechanistically, we discovered that the Qiangxin recipe promoted the activation of transcription factor Krüppel-like factor 5 (KLF5) to induce glucose metabolism and inhibit apoptosis in cardiomyocytes. Further, we identified that KLF5 increased the promoter activity of hexokinase 2 (HK2) and B-cell CLL/lymphoma 2 (BCL2) genes, which further enhanced glucose metabolism and inhibited apoptosis of cardiomyocytes. CONCLUSIONS: We highlighted the importance of KLF5-mediated signaling pathways in the treatment of CHF as shown by their participation in glucose metabolism and apoptosis in a doxorubicin-induced model of cardiomyocyte injury, as well as show that Qiangxin recipe can be used as a novel targeted therapy for the treatment of CHF. Compared with previous studies, we provide new ideas for the treatment of Doxorubicin-induced CHF from the perspective of energy metabolism.

Prospects of Probiotic Adjuvant Drugs in Clinical Treatment.

In modern society, where new diseases and viruses are constantly emerging, drugs are still the most important means of resistance. However, adverse effects and diminished efficacy remain the leading cause of treatment failure and a major determinant of impaired health-related quality of life for patients. Clinical studies have shown that the disturbance of the gut microbial structure plays a crucial role in the toxic and side effects of drugs. It is well known that probiotics have the ability to maintain the balance of intestinal microecology, which implies their potential as an adjunct to prevent and alleviate the adverse reactions of drugs and to make medicines play a better role. In addition, in the past decade, probiotics have been found to have excellent prevention and alleviation effects in drug toxicity side effects, such as liver injury. In this review, we summarize the development history of probiotics, discuss the impact on drug side effects of probiotics, and propose the underlying mechanisms. Probiotics will be a new star in the world of complementary medicine.

Effect of prior drought and heat stress on Camellia sinensis transcriptome changes to Ectropis oblique (Lepidoptera: Geometridae) resistance.

Tea plants are continuously confronted with a wide range of biotic and abiotic stressors in the field, which can occur concurrently or sequentially. To elucidate the molecular mechanisms in responses to such individual and combined stresses, we used RNAseq to compare the temporal changes in the transcriptome of Camellia sinensis to Ectropis oblique Prout alone or in combination with exposure to drought and heat. Compared with the individual stress, tea plants exhibit significant differences in transcriptome profiles under the combined stresses. Additionally, many unique genes exhibited significant differences in expression in individual and combined stress conditions. Our research showed novel insights into the molecular mechanisms of E. oblique Prout resistance in tea plants and provided a valuable resource for developing tea varieties with broad spectrum stress tolerance.

Comparative study on the in vitro digestibility of chicken protein after different modifications.

The effects of tea polyphenols (TPPs) and ultrasound treatment (UDT) on the digestibility of chicken myofibrillar protein (MPN) in anenhanced oxidation system were investigated. As observed, the original aggregates of MPN were much lower in the UDT-assisted group than in the control protein group, and the difference widened after the incorporation of TPPs. The covalent structures of the UDT-assisted oxidation groups were verified via mass spectrometry and amino acid (AAD) measurements. The peptide abundance increased after the UDT-assisted covalent reaction and most of these peptides were derived from the structural proteins of MPNs according to the results of nano-LC-ESI-MS/MS. Digestion kinetic analysis showed that the digestion level of the EGCG-treated group was better than that of the other treated groups, regardless of the UDT-assisted covalent reaction. Overall, the combination of EGCG oxidation and UDT may be an efficient way to promote the nutritional value of the final MPN products.

Efficacy and safety of laser combination therapy and laser alone therapy for keloid: a systematic review and meta-analysis.

To evaluate the efficacy and safety of laser alone therapy and laser combination therapy (mainly combined with other kinds of laser or steroids) for keloid.PubMed, Embase and Web of Science were searched for relevant articles from inception to June 2020. Comprehensive Meta-Analysis software 2.0 (CMA) was used to perform the meta-analysis.A total of 29 articles were included in this meta-analysis. During the mean follow-up of 14 (1-84) months, the overall improvement rates of baseline Vancouver scar scale (VSS) score and itch were 0.454 (95%CI 0.351-0.561, I2 = 0) and 0.786 (95%CI 0.613-0.895, I2 = 0) in the laser combination therapy group. The improvement rates of scar height and flexibility in the laser combination therapy group were 0.629 (95%CI 0.519-0.727, I2 = 52.089) and 0.784 (95%CI 0.251-0.975, I2 = 89.420). The average improvement rate of the scar score in laser combination therapy was 0.338 (0.201-0.510); however, there were insufficient data for laser alone therapy comparison. The laser combination therapy had a greater pain improvement rate, 0.580 (0.389-0.750) versus 0.420 (0.224-0.645), compared to laser alone therapy, and a greater degree of good or excellent (> 50%) improvement in the overall scar, 0.636 (95%CI 0.347-0.852) versus 0.149 (95%CI 0.032-0.482), with laser alone therapy. Moreover, a lower regrowth rate of 0.187 (0.129-0.263) versus 0.249 (0.060-0.631), a lower post-treatment pigmentation rate of 0.125 (0.091-0.169) versus 0.135 (0.058-0.282), and a lower infection rate of 0.047 (0.009-0.209) versus 0.076 (0.012-0.351) were observed in the laser combination therapy compared with those rates in the laser alone therapy.The overall effect of laser combination therapy was better than that of laser alone therapy, and the incidence of adverse reactions was lower in laser combination therapy than in laser alone therapy.

Camellia nitidissima Chi leaf as pancreatic lipase inhibitors: Inhibition potentials and mechanism.

In this study, Camellia nitidissima Chi leaf extract was investigated for its compounds and pancreatic lipase inhibitory potentials. The interaction was determined using ultraviolet (UV) spectroscopy, circular dichroism (CD), fluorescence spectroscopy (FS), and molecular docking to understand the inhibiton, kinetic, and conformation of extraction-pancreatic lipase complex. C. nitidissima Chi leaf extraction inhibited the pancreatic lipase activity in a dose-dependent manner at the concentration of 1-12 mg/ml. The Lineweaver-Burk plots indicated that the inhibition on pancreatic lipase by extraction was noncompetitive. In addition, the decrease in α-helix contents, increase in ß-sheet and ß-turn, and decrease in fluorescence intensity after extraction treatment indicated that the conformation of pancreatic lipase was changed. This work revealed that C. nitidissima Chi leaf extraction played a significant role in inhibiting pancreatic lipase activity and brought out a solution of delay fat accumulation. PRACTICAL APPLICATIONS: This study reports the components in the extract of C. nitidissima Chi leaf and its inhibitory effect and mechanism of pancreatic lipase. C. nitidissima Chi leaf is a good source of bioactive components, including multiflorin B, kaempferol-3-O-rutinoside, vicenin-2, apigenin-6-C-pentosyl-8-C-hexosyl, vitexin, kaempferol, and other ingredients. It can inhibit pancreatic lipase and be used to control obesity and treat hyperlipidemia. This study also revealed the structure changes of C. nitidissima Chi leaf extract on pancreatic lipase, and further revealed the inhibitory mechanism of C. nitidissima Chi leaf extract on lipase, which provides a theoretical basis for C. nitidissima Chi leaf as a lipase inhibitor.

Effects of cinnamon essential oil on the physical, mechanical, structural and thermal properties of cassava starch-based edible films.

The edible films were mainly made from oxidized hydroxypropyl cassava starch incorporated with cinnamon essential oil (CEO). The effects of CEO amount on the physical and mechanical properties of films were studied, and the structures of films with and without CEO were characterized. The results showed that the elongation at break, water resistance, water vapor transmission coefficient, as well as oxygen and ultraviolet barrier properties of the films (p < 0.05) significantly increased with addition of CEO, while the tensile strength of the films decreased. The field emission scanning electron microscopic (FE-SEM) images and infrared (IR) spectra showed that the CEO had good compatibility with other components and could be evenly dispersed in the film, which was conducive to the stable release of the active components. X-ray diffraction (XRD) patterns showed that the addition of CEO increased the crystallinity of the film, indicating that the compatibility and structural stability of the crystal structure of the film were improved. The thermogravimetric analysis results showed that CEO was beneficial to improve the thermal stability of the films. This study provided a potential to develop edible films from modified cassava starch with CEO.

Differential effects of EPA and DHA on DSS-induced colitis in mice and possible mechanisms involved.

BACKGROUND: The anti-inflammatory effect of n-3 PUFAs has been widely documented. Emerging evidence suggests that the main component of n-3 PUFAs, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may have differential effects in ulcerative colitis (UC). It was aimed to clarify their differential effects in UC. METHODS: Eight-week-old male C57BL/6J mice were randomly divided into 7 groups, namely control, UC model, salicylazosulfapyridine (SASP), low-dose DHA, high-dose DHA, low-dose EPA, and high-dose EPA. DHA, EPA and SASP treatment groups were orally treated accordingly for 9 weeks. During the 5th to 9th week the control group was given distilled water, while other groups were given distilled water with 2% dextran sodium sulfate (DSS) to induce UC. Body weight loss, diarrhea, and stool bleeding were recorded to calculate the disease activity index (DAI). The level of tight junction proteins Claudin-1 and Occludin, and cytokines including TNF-α, IL-6, and IL-1ß as well as inflammatory cell markers such as MPO, F4/80, and MCP-1 in the intestinal epithelium were measured using western blotting. Activation of IL-6/STAT3 and NLRP3/IL-1ß inflammatory pathways was also assessed. Levels of proliferation-related proteins of the Wnt/ß-catenin pathway with c-myc, Cyclin-D1, and PCNA were detected. RESULTS: EPA, superior to DHA, significantly attenuated DSS-induced colitis evidenced by reduced DAI scores, cytokine production and inflammatory cell infiltration. Mechanically, EPA triggered a marked up-regulation of Claudin-1 and Occludin with down-regulation of their up-stream Akt and ERK. EPA also inhibited NLRP3/IL-1ß and IL-6/STAT3 inflammatory pathways and up-regulated the Wnt/ß-catenin pathway. CONCLUSIONS: EPA is more suitable to be used for the treatment of UC than DHA.

Effects of different preheat treatments on volatile compounds of camellia (Camellia oleifera Abel.) seed oil and formation mechanism of key aroma compounds.

In this study, volatile compounds of camellia seed oil (CSO) prepared by different preheat treatments (microwave, frying, roasting, and steaming) were identified by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS). A total of 107 volatile compounds were identified in CSO samples, including aldehydes (16), alcohols (6), ketones (3), heterocyclic compounds (26), esters (23), hydrocarbons (15), and others (17). Among them, untreated CSO is mainly hydrocarbons, roasting and steaming CSO are mainly aldehydes and alcohols, while microwave and roasting CSO are dominated by aldehydes and heterocyclic compounds. Fourteen volatile compounds with high relative odor activity value (ROAV ≥ 1) were selected as key aroma compounds (KACs). Principal Component Analysis (PCA) and Cluster Analysis (CA) were performed on 14 KACs, which determined that there were 3, 3, 3, 7, and 6 characteristic aroma compounds (CACs) in untreated, microwaved, frying, roasting, and steaming CSO. Additionally, the potential formation pathways and mechanism of KACs were discussed. PRACTICAL APPLICATIONS: Flavor is an important factor for consumers to choose edible oils, and it is also one of the indicators of oil quality. Different flavors of CSO can cater to the needs of different consumers. CSO manufactories can choose different preheat treatments to produce CSO with various flavors to meet different customers' need. CSO with new flavor can extend its market share and increase its value.

Dingxin Recipe IV attenuates atherosclerosis by regulating lipid metabolism through LXR-α/SREBP1 pathway and modulating the gut microbiota in ApoE-/- mice fed with HFD.

ETHNOPHARMACOLOGICAL RELEVANCE: Dingxin Recipe (DXR) is a traditional Chinese medicine formula that has been reported to be effective and safe treatment for cardiovascular diseases, such as arrhythmias, coronary heart disease. Dingxin Recipe IV (DXR IV) was further improved from the DXR according to the traditional use. However, the mechanism of DXR IV in atherosclerosis is unclear. AIM OF THE STUDY: This study aimed to illustrate whether DXR IV improve atherosclerosis through modulating the lipid metabolism and gut microbiota in atherosclerosis mice. MATERIALS AND METHODS: 40 male ApoE-/- mice were fed on HFD for 12 weeks and were then treated with DXR IV (1.8, 0.9, or 0.45 g/kg/d) for another 12 weeks. The decroation of DXR IV contains four traditional Chinese medicines: the dried rhizome of Coptis chinensis Franch. (15.09%), the root of Salvia miltiorrhiza Bunge (28.30%), the seed of Ziziphus jujuba Mill. (37.74%) and the fruiting body of Ganoderma lucidum (Leyss.ex Fr.) Karst. (18.87%). 8 male c57BL/6 mice fed a normal diet served as control group. The atherosclerotic plaque was quantified by oil-red O staining and masson trichrome staining. Mice feces were collected. The gut micobiota were detected by 16S rRNA gene sequencing and fecal metabolites were analyzed by 1H NMR spectroscopy. The effect of DXR IV on blood lipids (TG, TC, LDL-C, HDL-C) was investigated. The lipid metabolism related genes were determined by RT-qPCR and western blotting respectively. RESULTS: DXR IV exerted the anti-atherosclerosis effect by inhibiting the excessive cholesterol deposition in aorta and regulating the level of TG, TC, LDL-C and HDL-C. The composition of gut microbiota was changed. Interestingly, the relative abundance of Muribaculaceae and Ruminococcaceae increased after DXR IV administration, whereas the abundance of Erysipelotrichaceae decreased, which have been beneficial to lipid metabolism. Nine potential metabolic biomarkers, including acetate, butyrate, propionate, alanine, succinate, valerate, xylose, choline, glutamate, were identified, which were related to fatty acid metabolism. Further, the pathway of fatty acid was detected by the RT-qPCR and western blotting. Compared with model group, the level of LXR-α and SREBP1 decreased significantly in DXR IV group while LXR-ß, SREBP2 showed no statistical significance. It indicated that DXR IV modulated lipid metabolism by LXR-α/SREBP1 but not LXRß and SREBP2. CONCLUSIONS: DXR IV exhibits potential anti-atherosclerosis effect, which is closely related to lipid metabolism and the gut microbiota. This study may provide novel insights into the mechanism of DXR IV on atherosclerosis and a basis for promising clinical usage.

Shen-Ling-Bai-Zhu-San alleviates functional dyspepsia in rats and modulates the composition of the gut microbiota.

The pathogenesis of functional dyspepsia (FD) is multifactorial, and the gut microbiota may play a significant role. Shen-Ling-Bai-Zhu-San (SLBZS), a traditional Chinese herbal medicine, has been widely used in the treatment of FD, and appears to influence the gut microbiota. Therefore, we hypothesized that SLBZS would alleviate dyspeptic symptoms by adjusting the composition of the gut microbiota. To test this hypothesis, we aimed to evaluate the effects of SLBZS on FD and elucidate the mechanism that underlies the interactions between gut microbiota and FD during SLBZS treatment. We employed a rat model of FD induced by multiple forms of chronic mild stimulation. 16S rRNA gene sequencing and shotgun metagenomic sequencing were used to analyze the microbial communities in fecal samples from the rats. We found that the SLBZS improved dyspeptic symptoms in FD rats, such as weight loss, decreased intestinal motility, reduced absorptive capacity. Moreover, the SLBZS treatment reversed gut dysbiosis in FD. With SLBZS treatment, FD biomarkers including Prevotella, Mucispirillum and Akkermansia were decreased while SCFA-producing bacteria such as Adlercreutzia and Clostridium, and sulfate-reducing bacteria Desulfovibrio were enriched. Additionally, SLBZS normalized the dysregulated function of the microbiome, upregulating the pathways of energy metabolism and decreasing the oxidative stress as well as bacterial pathogenesis. Our study demonstrated that SLBZS could ameliorate dyspepsia, and amend the dysregulated composition and function of the gut microbial community, providing insight into the mechanism of SLBZS treatment for FD from the perspective of gut microbiota.

The zinc-regulated protein (ZIP) family genes and glutathione s-transferase (GST) family genes play roles in Cd resistance and accumulation of pak choi (Brassica campestris ssp. chinensis).

The molecular mechanisms of the differences among the Cd tolerance and accumulation of different pak choi cultivars are essential to further breed Cd-safe genotypes pak choi. In our research, via morphological comparison, qRT-PCR and yeast function complementary approaches, we explored the differences of Cd tolerance and capacity for Cd uptake in nine various pak choi varieties. Results showed that higher expressions of BcZIPs involved in Cd uptake in 'Kang Re605' may lead to its higher capacity for Cd accumulation. The lowest expressions of transporter gene in 'Wu Yueman' were consistent with its fewest ability to uptake Cd. Beyond that, the difference of resistance was very great among varieties. Meanwhile, the expressions of the BcGSTUs were differentially induced by Cd exposure in different pak choi varieties, and 'Kang Re605' performed the highest BcGSTUs expression overall. To verify the role of GSTUs played in Cd resistance of pak choi, four BcGSTUs, BcGSTU4, BcGSTU11, BcGSTU12 and BcGSTU22 in a high-Cd accumulation and tolerance variety 'Kang Re605' were cloned, quantitated and transferred to Cd-sensitive yeast mutant strain. And finally found that BcGSTU11 increased the Cd tolerance of yeast, which may associate with a high Cd resistance of 'Kang Re605'. Simultaneously, less BcGSTUs abundance in 'Shang Haiqing' may result in its weak tolerance to Cd. These findings will help us to comprehend the roles of BcZIPs and BcGSTUs in Cd absorption and detoxification as well as promote our understanding of the Cd-resistant and Cd-accumulated mechanisms in pak choi.

Transcriptome analyses of Paris polyphylla var. chinensis, Ypsilandra thibetica, and Polygonatum kingianum characterize their steroidal saponin biosynthesis pathway.

Steroidal saponins, one of the most diverse groups of plant-derived natural products, elicit biological and pharmacological activities; however, the genes involved in their biosynthesis and the corresponding biosynthetic pathway in monocotyledon plants remain unclear. This study aimed to identify genes involved in the biosynthesis of steroidal saponins by performing a comparative analysis among transcriptomes of Paris polyphylla var. chinensis (PPC), Ypsilandra thibetica (YT), and Polygonatum kingianum (PK). De novo transcriptome assemblies generated 57,537, 140,420, and 151,773 unigenes from PPC, YT, and PK, respectively, of which 56.54, 47.81, and 44.30% were successfully annotated, respectively. Among the transcriptomes for PPC, YT, and PK, we identified 194, 169, and 131; 17, 14, and 26; and, 80, 122, and 113 unigenes corresponding to terpenoid backbone biosynthesis; sesquiterpenoid and triterpenoid biosynthesis; and, steroid biosynthesis pathways, respectively. These genes are putatively involved in the biosynthesis of cholesterol that is the primary precursor of steroidal saponins. Phylogenetic analyses indicated that lanosterol synthase may be exclusive to dicotyledon plant species, and the cytochrome P450 unigenes were closely related to clusters CYP90B1 and CYP734A1, which are UDP-glycosyltransferases unigenes homologous with the UGT73 family. Thus, unigenes of ß-glucosidase may be candidate genes for catalysis of later period modifications of the steroidal saponin skeleton. Our data provide evidence to support the hypothesis that monocotyledons biosynthesize steroidal saponins from cholesterol via the cycloartenol pathway.

[Correlation of MRI apparent diffusion coeffcient with molecular marker Ki-67 in gastric cancer].

OBJECTIVES: To study the correlation between MRI apparent diffusion coefficient (ADC) and expression of Ki-67 in gastric cancers, and to investigate the application of ADC value in diagnosing the malignance of gastric cancer. METHODS: A retrospective cohort analysis was performed on 87 gastric cancer patients who received MRI examination and radical resection at Zhejiang Provincial Hospital of Traditional Chinese Medicine from November 2014 to August 2015. All the postoperative resected samples were confirmed as gastric cancer. Preoperative MRI examination was performed by using Siemens 3.0-T Verio MRI with following parameters: section thickness 3 mm, gap 1 mm, matrix 182×320, field of view 40 cm. Plain scan was followed by T1-weighted fat suppression technique VIBE 3D(TR3.92/TE1.39,90degree) scans at arterial phase (the 30th second), portal venous phase (the 60th second), lag period (the 90th second), axial planes and coronal planes (the 180th second), and sagittal planes (the 210th second), respectively. ADC value of tumor was measured at b-factor of 800 s/mm2 and ADC map was generated from DWI data on the work station. The expression of Ki-67 in cancer tissue was detected by routine immunohistochemical (SP) staining after surgery. Correlation between ADC value and the expression of Ki-67 in gastric cancer was analyzed. RESULTS: Irregular thickening of the gastric wall and inhomogeneous enhancement of the tumor after injection of the contrast material appeared in gastric cancer. Gastric cancer tissue presented hyperintensity and normal gastric wall presented isointensity in DWI image (b=800 s/mm2). Compared with normal gastric tissue, mean ADC value of gastric cancer tissue was significant lower [(1.114±0.265)×10-3 mm2/s vs. (2.032±0.202)×10-3 mm2/s, t=26.209, P=0.000]. The ADC values of high-middle differentiation group, middle-low differentiation group, low differentiation group and signet ring cell carcinoma/mucinous adenocarcinoma group were (1.347±0.234)×10-3 mm2/s, (1.179±0.257)×10-3 mm2/s, (0.996±0.185)×10-3 mm2/s and (1.082±0.230)×10-3 mm2/s, respectively. The difference of mean ADC value among different tumor stages was significant(F=8.498, P=0.000). Along with the Ki-67 expression up-regulated, the ADC value decreased in cancer tissue. The Ki-67 expressions in cancer tissue was negatively correlated with cancer ADC values (r=-0.570, P=0.000). Furthermore, negative correlations of Ki-67 expressions with ADC values of high-middle differentiation group (r=-0.627, P=0.016), low differentiation group (r=-0.787, P=0.000) and signet ring cell carcinoma/mucinous adenocarcinoma group (r=-0.792, P=0.000) were observed respectively, while Ki-67 expression was not correlated with ADC value of middle-low differentiation group. CONCLUSION: The ADC value of gastric cancer can reflect the level of tumor differentiation, and is negatively correlated with Ki-67 expression in cancer tissues.