Expansin SlExp1 and endoglucanase SlCel2 synergistically promote fruit softening and cell wall disassembly in tomato.

Fruit softening, an irreversible process that occurs during fruit ripening, can lead to losses and waste during postharvest transportation and storage. Cell wall disassembly is the main factor leading to loss of fruit firmness, and several ripening-associated cell wall genes have been targeted for genetic modification, particularly pectin modifiers. However, individual knockdown of most cell wall-related genes has had minimal influence on cell wall integrity and fruit firmness, with the notable exception of pectate lyase. Compared to pectin disassembly, studies of the cell wall matrix, the xyloglucan-cellulose framework, and underlying mechanisms during fruit softening are limited. Here, a tomato (Solanum lycopersicum) fruit ripening-associated α-expansin (SlExpansin1/SlExp1) and an endoglucanase (SlCellulase2/SlCel2), which function in the cell wall matrix, were knocked out individually and together using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9-mediated genome editing. Simultaneous knockout of SlExp1 and SlCel2 enhanced fruit firmness, reduced depolymerization of homogalacturonan-type pectin and xyloglucan, and increased cell adhesion. In contrast, single knockouts of either SlExp1 or SlCel2 did not substantially change fruit firmness, while simultaneous overexpression of SlExp1 and SlCel2 promoted early fruit softening. Collectively, our results demonstrate that SlExp1 and SlCel2 synergistically regulate cell wall disassembly and fruit softening in tomato.

Tubson-2 decoction ameliorates rheumatoid arthritis complicated with osteoporosis in CIA rats involving isochlorogenic acid A regulating IL-17/MAPK pathway.

BACKGROUND: Osteoporosis (OP) is considered as one of the major comorbidities of rheumatoid arthritis (RA), and is responsible for fragility fracture. However, there is currently no effective treatment for RA complicated with OP. Tubson-2 decoction (TBD), a Mongolian medicine also known as Erwei Duzhong Decoction, has been shown to exert a preventive effect on post-menopausal osteoporosis (PMOP). The preventive effects of TBD on RA-induced OP, as well as the bioactive compound responsible and the underlying mechanisms, remain to be elucidated. OBJECTIVE: To explore the effects of TBD on RA-induced OP in vivo, and to elucidate the mechanism of isochlorogenic acid A (ICA), the effective component of TBD, in vitro. METHODS: To evaluate the anti-arthritic and anti-osteoporotic effects of TBD, we conducted H&E straining and safranine O/fast green, TEM, immunohistochemistry (IHC), bone histomorphometry, micro-CT imaging, and biomechanical testing in collagen induced arthritis (CIA) rats. The active ingredient in TBD was identified using network pharmacology and molecular docking. The identification was supported by in vivo IHC assay, and further confirmed using qRT-PCR, Western blot, and SEM analysis in TNF-α-treated MH7A cells and/or in LPS-exposed RAW264.7 cells. RESULTS: Oral administration of TBD attenuated the severity of arthritis and osteopenia as well as poor bone quality, in CIA rats. Additionally, TBD and the positive control, tripterygium glycosides (TG), exhibited similar effects in reducing inflammation in both the synovium and ankle joint. They also were both effective in improving bone loss, microarchitecture, and overall bone quality. TBD reduced the expression of MMP13, IL-17, and p-JNK protein in the synovium of CIA rats. ICA, which was screened, suppressed TNF-α or LPS-triggered inflammatory responses via down-regulating IL-17 signaling, involving in MMP13, IL-1ß, IL-23, and IL-17, and the MAPK pathway including p-ERK, p-JNK, and p-P38, both in MH7A cells and in RAW264.7 cells. Furthermore, ICA prevented osteoclasts from differentiating and bone resoprtion in a dose-dependent manner in vitro. CONCLUSION: This study provides the first evidence that TBD exerts intervening effects on RA-induced OP, possibly through the downregulation of the IL-17/MAPK signaling pathway by ICA. The findings of our study provides valuable insights for further research in this area.

Effect of intranasal acupuncture on the pathology of nasal mucosa in rabbits with allergic rhinitis and related regulatory mechanism of transient receptor potential vanillic acid receptor 1-substance P axis / 解剖学报

Objective To investigate the pathological changes of nasal mucosa by intranasal acupuncture, and to explore the possible signal transduction mechanism through the distribution and expression of transient receptor potential vanillic acid receptor 1 (TRPV1)-substance P (SP) axis in nasal mucosa of experimental allergic rhinitis. Methods An animal model of allergic rhinitis was constructed by ovalbumin and aluminium hydroxide gel treatment. Twenty five New Zealand rabbits were randomly divided into normal group, model group, sham acupuncture group, acupuncture Wai Ying Xiang group and intranasal acupuncture group, with 5 rabbits in each group. To observe the effect of intranasal acupuncture on animal behavior, nasal mucosa pathology, eosinophils (EOS) count and IgE content in arterial blood. In addition, HE staining was used to determine the distribution of EOS in nasal mucosa and immunohistochemistry was used to observe the distribution and expression of TRPV1 and SP in nasal mucosa membrane. Results Compared with the model group, the behavioral score of the sham acupuncture group did not decrease significantly, while the behavioral score of the Wai Ying Xiang acupuncture group and the intranasal acupuncture group showed a downward trend, and the decrease degree of the intranasal acupuncture group was more significant. Compared with the normal group, the eosinophil count in the model group increased slightly (P0.05). HE staining found that eosinophilic distribution increased significantly in the model group and the sham acupuncture group compared with the normal group, and the difference was statistically significant (P<0.05). Compared with the model group, the distribution of eosinophil reduced in the nasal acupuncture group and the Wai Ying Xiang group. The difference was statistically significant (P<0.05 ). Immunohistochemical found that TRPV1 and SP expressed significantly in the nasal mucosa of the model group and the sham acupuncture group compared with the normal group, the difference was statistically significant (P<0.05). Compared with the model group, TRPV1 and SP expression decreased significantly in the intranasal acupuncture group and the external yingxiang group, the difference was statistically significant (P<0.05). Conclusion Intranasal acupuncture can alleviate the nasal symptoms of allergic rhinitis model by reducing the Chemotaxis of eosinophils in nasal mucosa and the content of IgE in serum.

Targeted copper supplementation oriented theranostic for fluorescence and 19F NMR detection of tumors.

Alteration of the levels of copper is a promising approach for cancer therapy. Herein, we develop a dual-mode copper vehicle, M985. The biotin-tailed M985 can exert tumor-directed copper supplementation and undergo self-immolative cleavage in living cancerous cells, resulting in the liberation of F542 along with the generation of excess reactive oxygen species. Thus, fluorescence and 19F NMR detection is realized to specifically discriminate cancer cells. F542 acts as a fluorescence reporter and a potent cytotoxic agent, facilitating the visualization of molecular release and distribution, as well as confirming the ER autophagy-induced apoptosis. Therefore, we present a promising dual-mode theranostic M985 for the efficient detection and therapy of cancer.

Near-infrared laser-induced phase-shifted nanoparticles for US/MRI-guided therapy for breast cancer.

Breast cancer is a refractory malignancy particularly in women, with an accruing morbidity and mortality worldwide. The purpose of this study was to evaluate the efficacy of laser-induced near-infrared (NIR) PFP/Gd-DTPA/ICG@PLGA NPs (PGINPs) in US/MR imaging and regimen for breast cancer; gadolinium-DTPA (Gd-DTPA), perfluoropentane (PFP) and indocyanine green (ICG) were wrapped in the poly (lactic-co-glycolic) acid (PLGA) shell membrane via a double emulsion approach. In this study, under the irradiation of NIR laser, the ICG enriched in the cancerous tissue not only converted optic energy into thermal energy to rapidly heat up the cancer focus but also convert O2 to singlet oxygen (1O2), which can effectively destroy the cancer tissues through photothermal therapy (PTT) and photodynamic therapy (PDT). Meanwhile, the thermal energy thus generated could promote the gasification of PFP to enable visualization of cancer tissues under US imaging. Gd-DTPA in combination with US imaging can also significantly enhance MR imaging to provide US/MR dual-modal imaging. This study proves the efficacy of NIR-inspired multifunctional nano-system PGINPs to potentiate US/MR imaging and regimen for breast cancer.

A comprehensive genomic and growth proteomic analysis of antitumor lipopeptide bacillomycin Lb biosynthesis in Bacillus amyloliquefaciens X030.

Lipopeptides (such as iturin, fengycin, and surfactin) from Bacillus possess antibacterial, antifungal, and antiviral activities and have important application in agriculture and pharmaceuticals. Although unremitting efforts have been devoted to improve lipopeptide production by designing gene regulatory circuits or optimizing fermentation process, little attention has been paid to utilizing multi-omics for systematically mining core genes and proteins during the bacterial growth cycle. Here, lipopeptide bacillomycin Lb from new Bacillus amyloliquefaciens X030 was isolated and first found to have anticancer activity in various cancer cells (such as SMMC-7721 and MDA-MB-231). A comprehensive genomic and growth proteomic analysis of X030 revealed bacillomycin Lb biosynthetic gene cluster, key enzymes and potential regulatory proteins (PerR, PhoP, CcpA, and CsfB), and novel links between primary metabolism and bacillomycin Lb production in X030. The antitumor activity of the fermentation supernatant supplemented with amino acids (such as glutamic acid) and sucrose was significantly increased, verifying the role of key metabolic switches in the metabolic regulatory network. Quantitative real-time PCR analysis confirmed that 7 differential expressed genes exhibited a positive correlation between changes at transcriptional and translational levels. The study not only will stimulate the deeper and wider antitumor study of lipopeptides but also provide a comprehensive database, which promotes an in-depth analysis of pathways and networks for complex events in lipopeptide biosynthesis and regulation and gives great help in improving the yield of bacillomycin Lb (media optimization, genetic modification, or pathway engineering).

The food safety of DP-356Ø43 soybeans on SD rats reflected by physiological variables and fecal microbiota during a 90-day feeding study.

Soybean is an important food resource for the eastern countries and herbicide-tolerant genetically modified soybeans (GMS) were widely developed to deal with weeds problems. Unprocessed soybean flour instead of dehulled and defatted soybean meal was used to reflect the safety of soybean food in whole. Rats were given formulated diets containing DP-356Ø43 or non-GM soybean JACK at an incorporation rate of 7.5%, 15%, or 30% (w/w), respectively for 90 days. Targeted traditional toxicological response variables were measured to reflect the holistic health of animals. No treatment-related adverse or toxic effects were observed based on an examination of the daily clinical signs, body weight, food consumption, hematology, serum biochemistry, and organ weight or based on gross and histopathological examination. The results demonstrate that the soybean DP-356Ø43 is as safe for consumption as conventional soybean JACK. In the current study, the effect of a herbicide-tolerant GMS DP-356043 on identified intestinal microbiota was evaluated in a rodent feeding study compared with its conventional control JACK. Feces samples from rats consuming different diets were collected before the start of the experiment (time 0) and at monthly intervals (at the end of the 1st, 2nd and 3rd months) over the course of 90 days. Six types of bacterias shared by humans and rats were detected with Q-PCR. The results of QPCR indicated that the GMS 356Ø43 had a comparable effect on the abundance of Bifidobacterium group, Clostridium perfringens subgroup, Escherichia coli, and Bacteroides-Prevotella group as the non-GMS JACK.

Effects of Pulsatile Cupping on Body Pain and Quality of Life in People with Suboptimal Health:A Randomized Controlled Exploratory Trial.

Objective: The curative effect of pneumatic pulsatile cupping on pain has been shown. This study was conducted to investigate effects of the pulsating frequency of pneumatic pulsatile cupping, compared with traditional cupping (TC), on body pain and quality of life (QoL) in people with suboptimal health status (SHS). Materials and Methods: Ninety-six participants with SHS were randomized to low-frequency (LF; n = 24) or high-frequency (HF; n = 24) pulsating cupping, traditional cupping (TC; n = 24), or wait-list (WL; n = 24) groups. The LF, HF, and TC groups received 4 sessions of cupping over 2 weeks. Visual analogue scale (VAS; 0-100 mm) pain level and Short-Form-36 (SF-36) QoL measurements were taken before and after the intervention. Results: Both LF and HF reduced pain significantly (VAS: -28.26; 95% confidence interval [CI] -36.18 to -20.34; and -31.88, 95% CI -39.81 to -23.96; both P = 0.000) and improved QoL more than WL (SF-36, Bodily Pain dimension: 1.46, 95% CI: 0.85 to 2.07; and 1.75, 95% CI: 1.14 to 2.36, both P = 0.000). Compared to TC, LF and HF significantly reduced pain (VAS: -7.92, 95% CI: -15.75 to -0.08, P LT = 0.048; and -11.54, 95% CI: -19.38 to -3.70, P HT = 0.004) and improved QoL (SF-36, Bodily Pain dimension: 0.61, 95% CI: 0.01 to 1.21, P LT = 0.046; and 0.90, 95% CI: 0.30 to 1.50, P HT = 0.004). There was no significant difference between LF and HF. Conclusions: This study showed that, in patients with SHS, pulsatile cupping therapy could have a more-favorable effect to relieve body pain, compared to TC. LF and HF pulsation produced equivalent pain relief. Further studies investigating the underlying mechanism are needed. Trial registration: This trial was registered in the Chinese Clinical Trial Registry (ChiCTR-INR-16009345).

Histopathological nature of myofascial trigger points at different stages of recovery from injury in a rat model.

OBJECTIVE: To investigate the histopathological nature of myofascial trigger points (MTrPs) or spots (MTrSs) at different stages of recovery from injury in a rat model. METHODS: Forty Sprague-Dawley rats were randomly divided into two groups: a control group (CG) and experimental group (EG). The CG was further randomly subdivided into CG1 and CG2 subgroups. The CG2 was used for palpating the taut band and CG1 as a blank. EG was subdivided into three groups according to recovery times: 4 weeks (4W), 8 weeks (8W) and 12 weeks (12W); these groups consisted of eight rats each. All CG rats received no intervention, whereas the intervention in EG rats was by a blunt strike to the vastus medialis and eccentric exercise for 8 weeks. The taut bands with spontaneous electrical activity were then detected in the muscle to guide a muscle biopsy. The histopathological findings were investigated under optical and electron microscopes in all groups. RESULTS: Under optical microscopy, the differently augmented sizes of round fibres (contracture knots) with deep staining in the transverse section and fusiform shapes in a longitudinal view were clearly seen in CG2 and EGs with a large diameter; the number of contracture knots was significantly more in EGs than in CGs. Under an electron microscope, the mitochondria in EGs significantly decreased with abnormal structures. The sarcomeres were significantly shortened in the 8W and 12W EGs. CONCLUSION: An injury can cause activation of MTrSs in a muscle and an activated level of MTrPs depending on the number of contracture knots in muscle with impaired energy production.

[Clinical Trial of Acupuncture Treatment of Gastro-esophageal Reflex Disease by Needling Dorsal Segment of the Governor Vessel].

OBJECTIVE: To observe the curative effect of needling stimulation of the dorsal segment (7 spots below the spinous processes from T3 to T9) of the Governor Vessel for gastro-esophageal reflux disease (GERD). METHODS: A total of 60 cases of GERD patients were randomly divided into acupuncture group and medication group (n = 30 in each group). For patients of the acupuncture group, the depression spots below the spinous processes from T3 to T9, including both acupoints [Shenzhu (GV 12), Shendao (GV 11), Lingtai (GV 10), Zhiyang (GV 9), Jinsuo (GV 8)] and non-acupoints (T4, T8) were punctured with filiform needles, once every other day for 8 weeks. Patients of the medication group were asked to take Omeprazole capsules (20 mg/time) twice a day for 8 weeks. The curative effects were assessed by "Reflux Disease Questionnaire" score (RDQ score of heartburn, retrosternal pain, acid regurgitation and food regurgitation, 0-40 points, the "Guidelines for Clinical Research on Chinese New Herbal Drugs" (GRED symptom scale scores about the 38 symptoms of the digestive system, respiratory system, five-sense organs, cardiovascular system, back-pain, sleep, headache, daily life activity, etc. 0-114 points, 0-3 scales), and "the Consensus about Diagnosis and Treatment of GERD of Traditional Chinese Medicine", respectively. RESULTS: Following the treatment, both RDQ score and GERD symptom scale score were significantly decreased in patients of the acupuncture and medication groups (P<0.05), and the effect of acupuncture was markedly superior to that of the medication in lowering RDQ and GERD scale scores (P<0.05). The cured plus markedly effective rate for RDQ score and the effective rate for GERD scale score were 70.0% (21/30) and 93.3% (28/30) respectively in the acupuncture group, being significantly higher than those [43.3% (13/30) and 66.7% (20/30)] in the medication group (P<0.05). CONCLUSION: Acupuncture stimulation of the dorsal segment

[Andrographolide inhibits extracellular signal-regulated kinase 1/2 signaling pathway in activated macrophages].

OBJECTIVE: To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages. METHODS: LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay. RESULTS: Andrographolide at 1-100 µg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 µg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 µg/mL andrographolide and 20 µmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels. CONCLUSION: In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.