Electroacupuncture alleviates streptozotocin-induced diabetic neuropathic pain via suppressing phosphorylated CaMKIIα in rats.

Diabetic neuropathic pain (DNP) is a frequent complication of diabetes. Calcium/calmodulin-dependent protein kinase II α (CaMKIIα), a multi-functional serine/threonine kinase subunit, is mainly located in the surface layer of the spinal cord dorsal horn (SCDH) and the primary sensory neurons in dorsal root ganglion (DRG). Numerous studies have indicated electroacupuncture (EA) takes effect in various kinds of pain. In this research, we explored whether CaMKIIα on rats' SCDH and DRG participated in DNP and further explored the mechanisms underlying the analgesic effects of EA. The DNP model in rats was successfully established by intraperitoneal injection of streptozotocin. Certain DNP rats were treated with intrathecal injections of KN93, a CaMKII antagonist, and some of the DNP rats received EA intervention. The general conditions, behaviors, the expressions of CaMKIIα and phosphorylated CaMKIIα (p-CaMKIIα) were evaluated. DNP rats' paw withdrawal threshold was reduced and the expressions of p-CaMKIIα in SCDH and DRG were upregulated compared with the Normal group, while the level of CaMKIIα showed no significance. KN93 attenuated DNP rats' hyperalgesia and reduced the expressions of p-CaMKIIα. We also found EA attenuated the hyperalgesia of DNP rats and reduced the expressions of p-CaMKIIα. The above findings suggest that p-CaMKIIα in SCDH and DRG is involved in DNP. The analgesic effect of EA in DNP might be related to the downregulation of p-CaMKIIα expression level. Our study further supports that EA can be an effective clinical treatment for DNP.

The potential mechanism of Guizhi Fuling Wan effect in the treatment of cervical squamous cell carcinoma: A bioinformatics analysis investigation.

As a global malignancy with high mortality rate, targeted drug development for Uterine Cervical Neoplasms is an important direction. The traditional formula Guizhi Fuling Wan (GFW) is widely used in gynecological diseases. However, its potential mechanism of action remains to be discovered. We retrieved GFW and cervical squamous cell carcinoma (CSCC) targets from public databases. The protein-protein interaction network was obtained by string computational analysis and imported Cytoscape_v3.9.0 to obtain the core network and the top 10 Hub genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes were used for enrichment analysis of the core network, and then molecular docking to verify whether the selected signaling pathway binds well to the core node. Finally, clinical prognostic analysis and expression differences of Hub genes were validated using the Cancer Genome Atlas database and R language. Our search yielded 152 common targets for GFW and CSCC. The interleukin-17 signaling pathway, tumor necrosis factor signaling pathway, and Toll-like signaling pathway were then selected for further molecular docking from the hub genes enrichment analysis results, which showed good binding. Among the Hub genes, JUN, VEGFA, IL1B, and EGF had a poor prognosis for CSCC. In conclusion, this study illustrates that GFW can have adjuvant therapeutic effects on CSCC through multiple targets and multiple pathways, providing a basis for further research.

Life cycle assessment perspective on waste resource utilization and sustainable development: A case of glyphosate production.

The growing demand for pesticide manufacturing and increasing public awareness of sustainable development, have let to urgent requirements for a refined environmental management framework. It is imperative to conduct process-based life cycle assessments (LCAs) to promote clean and environment-friendly technologies. Herein, the cradle-to-gate LCA of glyphosate production was executed as an example to investigate crucial production factors (materials or energy) and multiple environmental impacts during the production processes. Results showed that methanol caused the highest environmental damage in terms of toxicity, with a normalized value of 85.7 × 10-8, followed by coal-fired electricity in 6.00 × 10-8. Furthermore, optimized schemes were proposed, including energy improvement (electricity generated by switching from coal-fired power to solar power) and wastewater targeted conversion. Regarding the normalization results before and after optimization, the latter showed more significant results with the normalized value decreasing by 21.10 × 10-8, while that of the former only decreased by 6.50 × 10-8. This study provides an integrated LCA framework for organophosphorus pesticides (OPs) from upstream control and offers an important supplement to managing the key pollution factors and control links of the OP industry. Moreover, it reveals the positive influence of optimized schemes in facilitating cleaner production technologies, thus ultimately promoting new methodologies for resource recycling.

Selenium application during fruit development can effectively inhibit browning of fresh-cut apples by enhancing antioxidant capacity and suppressing polyphenol oxidase activity.

Browning is a crucial factor affecting the quality of fresh-cut apples. A safe, simple, and effective method to inhibit browning is urgently needed in fresh-cut apple production. We carried out this study to explore the effect mechanism of exogenous selenium (Se) fertilizer on fresh-cut apple browning. During the development of apples, 0.75 kg/plant Se fertilizer was exerted on the 'Fuji' apple tree at the critical stage of the young fruit stage (late May), early fruit expansion stage (late June), and fruit expansion stage (late July), an equal amount of Se-free organic fertilizer was used as control. Polyphenol oxidase (PPO), peroxidase (POD), and phenylalanine ammonia-lyase (PAL) activities, phenolic and malondialdehyde (MDA) content, antioxidant enzymes activity, and DPPH free radical scavenging rate of the apple at different development stages were investigated. The highest Se accumulation efficiency was observed in apple fruit one month after applying Se fertilizer, which was 41.1%. Se-rich apples exhibited a more remarkable ability to resist browning than control after fresh-cut. The anti-browning effect of the fertilization group (M7) was the best, the PPO activity decreased to 0.5 × 103 U kg-1, and the browning index was 28.6. The total Se content (TSC) of 331.4 µg kg-1 DW and organic Se content (OSC) of 292.0 µg kg-1 DW were the highest in the apple samples, reached the classification standard of Se content in Se-rich food. The correlation analysis found that fresh-cut apple browning was closely related to antioxidant capacity and PPO activity. The stronger the antioxidant capacity of fresh-cut apples treated with Se fertilizer, the lower their browning degree. Therefore, exogenous Se can alleviate fresh-cut apples browning by improving antioxidant capacity and reducing PPO activity. Se-rich apples could increase the Se content of the human essential trace element and inhibit the browning of fresh-cut apples, which would become a new, safe and effective way to solve the fresh-cut apples browning.

Exploring the mechanisms of self-made Kuiyu Pingchang recipe for the treatment of ulcerative colitis and irritable bowel syndrome using a network pharmacology-based approach and molecular docking.

BACKGROUND: Ulcerative colitis (UC) and irritable bowel syndrome (IBS) are common intestinal diseases. According to the clinical experience and curative effect, the authors formulated Kuiyu Pingchang Decoction (KYPCD) comprised of Paeoniae radix alba, Aurantii Fructus, Herba euphorbiae humifusae, Lasiosphaera seu Calvatia, Angelicae sinensis radix, Panax ginseng C.A. Mey., Platycodon grandiforus and Allium azureum Ledeb. OBJECTIVE: The aim of the present study was to explore the mechanisms of KYPCD in the treatment of UC and IBS following the Traditional Chinese Medicine (TCM) theory of "Treating different diseases with the same treatment". METHODS: The chemical ingredients and targets of KYPCD were obtained using the Traditional Chinese Medicine Systems Pharmacology database and analysis platform (TCMSP). The targets of UC and IBS were extracted using the DisGeNET, GeneCards, DrugBANK, OMIM and TTD databases. The "TCM-component-target" network and the "TCM-shared target-disease" network were imaged using Cytoscape software. The protein-protein interaction (PPI) network was built using the STRING database. The DAVID platform was used to analyze the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Using Autodock Tools software, the main active components of KYPCD were molecularly docked with their targets and visualized using PyMOL. RESULTS: A total of 46 active ingredients of KYPCD corresponding to 243 potential targets, 1,565 targets of UC and 1,062 targets of IBS, and 70 targets among active ingredients and two diseases were screened. Core targets in the PPI network included IL6, TNF, AKT1, IL1B, TP53, EGFR and VEGFA. GO and KEGG enrichment analysis demonstrated 563 biological processes, 48 cellular components, 82 molecular functions and 144 signaling pathways. KEGG enrichment results revealed that the regulated pathways were mainly related to the PI3K-AKT, MAPK, HIF-1 and IL-17 pathways. The results of molecular docking analysis indicated that the core active ingredients of KYPCD had optimal binding activity to their corresponding targets. CONCLUSION: KYPCD may use IL6, TNF, AKT1, IL1B, TP53, EGFR and VEGFA as the key targets to achieve the treatment of UC and IBS through the PI3K-AKT, MAPK, HIF-1 and IL-17 pathways.

[Effect of functional acupoint electrical stimulation combined with tadalafil on erectile dysfunction in middle-aged and elderly patients].

OBJECTIVE: To investigate the application effect of functional acupoint electrical stimulation combined with tadara irregular administration in middle-aged and elderly patients with erectile dysfunction (ED), and to provide reference for clinical treatment. METHODS: A total of 40 middle-aged and elderly patients with ED admitted to the pelvic floor Center of our hospital from March 2021 to March 2023 were randomly divided into two groups with 20 cases in each group.The control group was treated with tadalafil regularly, and the observation group was treated with functional acupoint electrical stimulation on the basis of this treatment. The total course of treatment was 6 weeks.The clinical efficacy of the two groups was compared. The therapeutic efficacy was evaluated by the International Erectile Function Index (IIEF-5), penile hardness score (EHS), serum total testosterone (TT) level, sexual satisfaction scale (SS) and pelvic floor electromyography, and the occurrence of adverse events was recorded. RESULTS: The total effective rate of the observation group was significantly higher than that of the control group (90% vs 70%, P < 0.05). After 6 weeks of treatment, both groups showed improvements in IIEF-5, EHS, SS, and TT compared to before treatment (P < 0.01). However, the improvement in the observation group was significantly better than that in the control groupï¼»IIEF-5: (22.13±2.11) vs (19.69±2.04), EHS: (3.68±0.47) vs (2.89±0.60), SS: (77.41±7.59) vs (70.32±7.28), TT: (13.43±3.89) nmol/L vs (8.85±3.02) nmol/L, all P < 0.01ï¼½; There were no significant changes in pelvic floor muscle electromyography values in the control group before and after treatment (P > 0.05), while in the observation group, pelvic floor muscle electromyography values (PFMV) in the pre-resting phase, fast muscle (Type II muscle) phase, slow muscle (Type I muscle) phase, endurance testing phase, and post-resting phase all improved compared to before treatment and were superior to the control group (P < 0.05). CONCLUSION: Functional acupoint electrical stimulation combined with tadara irregular administration can improve the therapeutic effect of middle-aged and elderly patients with ED, improve pelvic floor function, safe and reliable.

Genome-wide investigation of the ZF-HD gene family in Tartary buckwheat (Fagopyrum tataricum).

BACKGROUND: ZF-HD is a family of genes that play an important role in plant growth, development, some studies have found that after overexpression AtZHD1 in Arabidopsis thaliana, florescence advance, the seeds get bigger and the life span of seeds is prolonged, moreover, ZF-HD genes are also participate in responding to adversity stress. The whole genome of the ZF-HD gene family has been studied in several model plants, such as Arabidopsis thaliana and rice. However, there has been little research on the ZF-HD genes in Tartary buckwheat (Fagopyrum tataricum), which is an important edible and medicinal crop. The recently published whole genome sequence of Tartary buckwheat allows us to study the tissue and expression profiles of the ZF-HD gene family in Tartary buckwheat on a genome-wide basis. RESULTS: In this study, the whole genome and expression profile of the ZF-HD gene family were analyzed for the first time in Tartary buckwheat. We identified 20 FtZF-HD genes and divided them into MIF and ZHD subfamilies according to phylogeny. The ZHD genes were divided into 5 subfamilies. Twenty FtZF-HD genes were distributed on 7 chromosomes, and almost all the genes had no introns. We detected seven pairs of chromosomes with fragment repeats, but no tandem repeats were detected. In different tissues and at different fruit development stages, the FtZF-HD genes obtained by a real-time quantitative PCR analysis showed obvious expression patterns. CONCLUSIONS: In this study, 20 FtZF-HD genes were identified in Tartary buckwheat, and the structures, evolution and expression patterns of the proteins were studied. Our findings provide a valuable basis for further analysis of the biological function of the ZF-HD gene family. Our study also laid a foundation for the improvement of Tartary buckwheat crops.

(+)-Usnic Acid Inhibits Migration of c-KIT Positive Cells in Human Colorectal Cancer.

Inhibition of tumor cell migration is a treatment strategy for patients with colorectal cancer (CRC). SCF-dependent activation of c-KIT is responsible for migration of c-KIT positive [c-KIT(+)] cells of CRC. Drug resistance to Imatinib Mesylate (c-KIT inhibitor) has emerged. Inhibition of mTOR can induce autophagic degradation of c-KIT. (+)-usnic acid [(+)-UA], isolated from lichens, has two major functions including induction of proton shuttle and targeting inhibition of mTOR. To reduce hepatotoxicity, the treatment concentration of (+)-UA should be lower than 10 µM. HCT116 cells and LS174 cells were employed to investigate the inhibiting effect of (+)-UA (<10 µM) on SCF-mediated migration of c-KIT(+) CRC cells. HCT116 cells were employed to investigate the molecular mechanisms. The results indicated that firstly, 8 µM (+)-UA decreased ATP content via uncoupling; secondly, 8 µM (+)-UA induced mTOR inhibition, thereby mediated activation suppression of PKC-A, and induced the autophagy of the completed autophagic flux that resulted in the autophagic degradation and transcriptional inhibition of c-KIT and the increase in LDH release; ultimately, 8 µM (+)-UA inhibited SCF-mediated migration of CRC c-KIT(+) cells. Taken together, 8 µM could be determined as the effective concentration for (+)-UA to inhibit SCF-mediated migration of CRC c-KIT(+) cells.

Effect of Chaiqin Chengqi Decoction on cholecystokinin receptor 1-mediated signal transduction of pancreatic acinar cells in acute necrotizing pancreatitis rats.

OBJECTIVE: To investigate the effect of Chaiqin Chengqi Decoction (,CQCQD) on cholecystokinin receptor 1 (CCKR1)-mediated signal transduction of pancreatic acinar cell in rats with acute necrotic pancreatitis (ANP). METHODS: Twenty-seven Sprague-Dawley rats were randomized into three groups: the control group, the ANP group, and the CQCQD group (9 in each group). ANP rats were induced by two intraperitoneal injections of 8% L-arginine (pH=7.0, 4.4 g/kg) over a 2-h period. Rats were treated with 1.5 mL/100 g body weight of CQCQD (CQCQD group) or physiological saline (control and ANP groups) at 2 h interval. And 6 h after induction, pancreatic tissues were collected for histopathological examination. Pancreatic acinar cells were isolated for determination of CCKR1 mRNA and protein expression, phospholipase C (PLC) and inositol-1,4,5-triphosphate (IP3), and determination of fluorescence intensity (FI) as a measure of intracellular calcium ion concentration [Ca(2+)]i. RESULTS: The pancreatic histopathological score (6.2 ± 1.1) and the levels of PLC (1,187.2 ± 228.2 µg/mL) and IP3 (872.2 ± 88.4 µg/mL) of acinar cells in the ANP group were higher than those in the control (2.8 ± 0.4, 682.5 ± 121.8 µg/mL, 518.4 ± 115.8 µg/mL) and the CQCQD (3.8 ± 0.8, 905.3 ± 78.5 µg/mL, 611.0 ± 42.5 µg/mL) groups (P<0.05). [Ca(2+)]i FI for the ANP group (34.8±27.0) was higher than that in the control (5.1 ± 2.2) and CQCQD (12.6 ± 2.5) groups (P<0.05). The expression of pancreatic acinar cell CCKR1 mRNA in the ANP group was up-regulated (expression ratio=1.761; P=0.024) compared with the control group. The expression of pancreatic acinar cell CCKR1 mRNA in the CQCQD group was down-regulated (expression ratio=0.311; P=0.035) compared with the ANP group. The ratio of gray values of the CCKR1 and ß-actin in the ANP group (1.43 ± 0.17) was higher than those in the control (0.70 ± 0.15) and CQCQD (0.79 ± 0.11) groups (P<0.05). CONCLUSIONS: Pancreatic acinar cell calcium overload of ANP induced by L-arginine was related to the up-regulated expressions of pancreatic acinar cell CCKR1 mRNA and protein. CQCQD can down-regulate expressions of pancreatic acinar cell CCKR1 mRNA and protein to reduce the PLC and IP3 of pancreatic acinar cells, relieving the calcium overload and reducing the pathological changes in rats with ANP.

Traditional Chinese medicine (Shun-Qi-Tong-Xie Granule) for irritable bowel syndrome: study protocol for a randomised controlled trial.

BACKGROUND: Irritable bowel syndrome (IBS) is a common gastrointestinal functional disorder with no effective therapy. Traditional Chinese medicine (TCM) is one of the most common complementary therapies in China. We designed this study to evaluate the efficacy and safety of Shun-Qi-Tong-Xie Granule (SQTX Granule), a TCM treatment, in patients with IBS with diarrhea (IBS-D). METHODS/DESIGN: A randomised, double-blinded, placebo-controlled, multi-centre, superiority clinical trial to evaluate the efficacy and safety of SQTX Granule is proposed. Eligible patients (Rome III) with IBD-S will be randomly assigned into SQTX Granule group and the placebo group. Patients will receive a 28-day treatment and a 2-month follow-up. The primary outcome measures include the scores of IBS-quality of life (IBS-QOL) rating scale and IBS-symptom severity scale (IBS-SSS) rating scale. The secondary outcome measures include the improvement of symptom scores, and the duration of abdominal pain and diarrhea. DISCUSSION: According to TCM theory, SQTX Granule has a regulating effect on abdominal pain, diarrhea and the syndrome of liver-spleen disharmony, which is similar to the symptoms of IBS-D. This study will provide objective evidence to evaluate the efficiency and safety of SQTX Granule in IBS-D treatment. TRIAL REGISTRATION: ChiCTR-TRC-14004241. Date of registration: 9 February 2014.

Artesunate exerts an anti-immunosuppressive effect on cervical cancer by inhibiting PGE2 production and Foxp3 expression.

Artesunate (ART), derived from a common traditional Chinese medicine, has beeen used an antimalarial for several years. In this study, the effect and mechanism of ART on anti-human cervical cancer cells was examined. The level of prostaglandin E2 (PGE2 ) and the population of CD4+CD25+Foxp3 regulatory T cells (Treg) in peripheral blood were detected by flow cytometry. In vivo antitumor activity was investigated in mice with cervical cancer by the subcutaneous injection of various concentrations of ART. The concentrations of PGE2 in the supernatants of CaSki cells were measured using an ELISA kit. Cyclooxygenase-2 (COX-2) and Foxp3 expression were determined using quantitative polymerase chain reaction (qPCR) and western blot analysis. The effect of ART on the viability of CaSki and Hela cells was evaluated with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. It was identified that the level of PGE2 and the population of CD4+CD25+Foxp3 Treg cells in the peripheral blood were significantly higher in cervical cancer patients and mice with cervical cancer. ART was capable of inhibiting orthotopic tumor growth, which correlated with a decrease in the level of PGE2 and the percentage of Treg cells in mice with cervical cancer. Furthermore, ART decreased COX-2 expression and the production of PGE2 in CaSki and Hela cells. Notably, the supernatants of CaSki cells treated with ART lowered the expression of Foxp3 in Jurkat T cells, which was capable of being reversed by exogenous PGE2 . Our data revealed that ART may elicit an anti-tumor effect against cervical cancer by inhibition of PGE2 production in CaSki and Hela cells, which resulted in the decrease of Foxp3 expression in T cells. Therefore, ART may be an effective drug for immunotherapy of cervical cancer.

Effect of H2O2/HCl heat treatment of implants on in vivo peri-implant bone formation.

PURPOSE: To investigate the effect of H2O/HCl heat treatment on peri-implant bone formation in vivo. MATERIALS AND METHODS: Twenty Ti-6Al-4V implants and 30 Ti-6Al-4V discs were used in this study. The implants and discs were separated into 2 groups: sandblasted and dual acid-etched group (control group) and sandblasted, dual acid-etched and H2O2/HCl heat-treated group (test group). Surface morphology, roughness, and crystal structure of the discs were analyzed by field-emission scanning electron microscopy, atomic force microscopy, and low angle X-ray diffractometry. The implants were inserted into the femurs of 10 adult white rabbits. Animals were injected with fluorescent bone labels at 1, 5, and 7 weeks following surgery to monitor progress of bone formation. Animals were euthanized 8 weeks postsurgery, and block biopsies were prepared for histologic and histometric analysis. RESULTS: Microscopic evaluation showed the surfaces were quite irregular for both techniques; however, the test surface demonstrated consistently smaller surface irregularities. The differences in Sa values were significant (P = .022). No significant differences were found in the maximum peak-to-valley ratio values (P = .258). X-ray diffractometry analysis showed that titanium dioxide was found on the test surface. New bone was formed on both implant surfaces. The bone-implant contact pattern appeared to produce a broad-based direct contact. Test implants demonstrated 7.13% more bone to implant contact (P = .003) and 15.42% more bone to implant contact for 3 consecutive threads (P = .001) than control implants. Test implants demonstrated 37.04% more bone area 500 microm outside of implant threads (P = .004) and 51.97% more bone area within 3 consecutive threads (P = .001) than control implants. No significant differences were found in bone area within all implant threads between the two groups (P = .069). CONCLUSION: This study demonstrated that implants heat-treated with H2O2/HCl solution enhanced peri-implant bone formation.