[Berberine alleviates programmed necrosis of metabolic-associated fatty liver disease via activating Nrf2 pathway in mice].

Objective: To investigate the effect of berberine on programmed necrosis of hepatocytes induced by metabolic-associated fatty liver disease (MAFLD) in mice and its related molecular mechanism. Methods: Twenty male C57BL/6N mice were randomly divided into four groups (n=5 in each group): control group (S), fatty liver group (H), berberine group(B), nuclear factor erythroid 2-related factor 2 inhibitor group (Nrf2), and all-trans-retinoic acid (ATRA) group (A). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), triglycerides (TG), total cholesterol (TC), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) concentrations were detected at the end of week 12 to calculate fatty liver index (liver mass/body mass ratio). Liver tissue was stained with HE, Masson and Oil Red O, and SAF score was used to evaluate the degree of liver injury. The expression levels of hepatic programmed necrosis-related proteins, namely receptor-interacting protein kinase 3 (RIPK3), phosphorylated mixed series protease-like domain (p-MLKL) and Nrf2 were detected by Western blot method. One-way ANOVA was used for intragroup comparisons and LSD-t tests were used for intergroup comparisons. Results: Compared with S group, H group serum ALT, AST, LDH, TG, TC, TNF-α, IL-1ß levels and fatty liver index were significantly increased. The liver tissue was filled with vacuolar-like changes and inflammatory cell infiltration. Numerous red lipid droplets were observed with oil red O staining. Collagen fiber hyperplasia was evident with Masson staining. SAF scores (6.60 ± 0.55 and 0.80 ± 0.45) were significantly increased. The expressions of RIPK3 and p-MLKL were up-regulated. Nrf2 level was relatively increased, and the differences were statistically significant (P < 0.05). Compared with H group, berberine intervention group liver biochemical indexes, lipid levels, pro-inflammatory mediator expression, fatty liver index, and SAF score were significantly reduced, and the expression of RIPK3 and p-MLKL were down-regulated, while Nrf2 levels were further increased, and the differences were statistically significant (P<0.05). Compared with B group, treatment with Nrf2 inhibitor had antagonized the protective effect of berberine on fatty liver. Serum ALT, AST, LDH, TG, TC and TNF-α, IL-1ß levels, fatty liver index, and SAF scores were significantly increased and the expressions of RIPK3 and p-MLKL were relatively increased, and the differences were statistically significant (P < 0.05). Conclusion: Berberine can significantly improve the metabolic-associated fatty liver disease injury in mice, and its mechanism is related to activation of Nrf2 and inhibition of programmed necrosis of hepatocytes.

[Epidemiological survey of mental disorders in the rural left behind elderly aged 60 years and older in Gansu].

Objective: To investigate the prevalence, demographic characteristics and social life function of mental disorders in the rural left behind elderly aged 60 years and older in Gansu. Methods: Between November 2017 and June 2018, a multi-stage stratified cluster sampling method was used to randomly select the rural left behind elderly aged 60 years and older in Gansu, and totally 6 000 elderly were enrolled. By using the extended general health questionnaire (GHQ-12) and the American Handbook for Diagnosis and Statistics of Mental Disorders (DSM-Ⅳ) Axis Ⅰ Disorders Formal Clinical Examination Patient Edition, all the included subjects were screened and diagnosed. Functional status was assessed by the Global Assessment Function scale (GAF). Statistical analysis of the prevalence of various mental illnesses, as well as the differences in the prevalence of different gender, marital status and age groups was performed. Results: Totally, 6 000 subjects completed the survey. The adjusted current prevalence of any mental disorder was 20.11% (95%CI 17.70%-22.85%). The six most prevalent specific disorders were major depressive disorder (9.20%), pain disorder (2.71%), mood disorder due to the body condition (2.08%), generalized anxiety disorder (1.99%), anxiety disorder not otherwise specified (1.15%) and dysthymic disorder (0.84%). The lifetime prevalence of mental disorders was 20.54% (95%CI 18.40%-23.39%). The overall current prevalence of mental disorders was higher in women (242.89‰) than in men (119.55‰), and the unmarried (248.37‰) was higher than those married (187.53‰). There was no significant difference in the prevalence of mental disorders among different age groups (P>0.05). The GAF score of mental disorders was 56±11, and 71.82% was moderate to severe functional impairment. Conclusions: The prevalence of mental disorders is high in rural left-behind population aged 60 years and over in Gansu Province. Major depression is a condition that deserves special attention.

Inhibition of TRPV1 prevented skin irritancy induced by phenoxyethanol. A preliminary in vitro and in vivo study.

BACKGROUND: Phenoxyethanol is a widely used preservative in personal care products. Transient receptor potential vanilloid 1 (TRPV1) on cell membrane is activated by TRPV1 agonist capsaicin resulting in an opening of the channel for calcium influx, which is linked with neurosensory sensations characterized by itching, burning and stinging of skin. Whether uncomfortable skin sensations caused by phenoxyethanol claimed by people having sensitive skin are also due to activation of TRPV1 has not been reported in the literature. OBJECTIVE: The aim of this study was to evaluate whether the TRPV1 is involved in the induction of itching and burning sensation by phenoxyethanol. METHODS AND MATERIALS: The effect of phenoxyethanol on TRPV1 was assessed in vitro on HaCaT cells. The activation of TRPV1 and its inhibition by a TRPV1 antagonist were evaluated by cellular calcium influx. TRPV1 protein expression was also investigated by Western blot. In vivo in a split-face study, phenoxyethanol formulated at 1% was compared to a formulation additionally containing a TRPV1 antagonist. By applying the formulations to the nasolabial fold, the scores of phenoxyethanol-induced sensations were compared to those of the TRPV1 antagonist. RESULTS: In vitro phenoxyethanol induced calcium influx in HaCaT cells in a dose-dependent manner after 20 min. This effect was abolished by a solution containing the TRPV1 antagonist trans-tert-butyl cyclohexanol (ID1609). Phenoxyethanol had no effect on the expression of TRPV1, whereas capsaicin caused a significant downregulation of this receptor in the same experiment. In vivo 1% phenoxyethanol induced a skin burning and itching sensation in a cohort of 60 of 243 Chinese female subjects being sensitive to phenoxyethanol discomfort. The uncomfortable skin sensations were significantly inhibited by ID1609. CONCLUSIONS: Different to capsaicin, phenoxyethanol did not downregulate the expression of TRPV1 in HaCaT cells, suggesting that different regulatory mechanisms may exist for capsaicin and phenoxyethanol. Our experiments demonstrated that phenoxyethanol induces skin misperception and uncomfortable skin sensations like itching and burning comparable to capsaicin via activating TRPV1. The stimulation was inhibited by blocking TRPV1 with ID1609. The present data strengthened hitherto studies that TRPV1 plays a critical role in sensitive skin.

Genetic diversity analysis of sweet kernel apricot in China based on SSR and ISSR markers.

Simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers were used to evaluate genetic diversity among 22 sweet kernel apricot accessions and 12 cultivars in China to provide information on how to improve the utilization of kernel apricot germplasms. The results showed that 10 pairs of SSR primers screened from 40 primer pairs amplified 43 allelic variants, all of which were polymorphic (100%), and 9 ISSR primers selected from 100 primers amplified 67 allelic variants with 50 polymorphic bands (74.63%). There was a relatively distant genetic relationship between the 34 samples, where their genetic similarity coefficient was between 0.62 and 0.99. The UPGMA dendrogram constructed using combined data of the two marker systems separated the genotypes into three main clusters.

Early strategy vs. late initiation of renal replacement therapy in adult patients with acute kidney injury: an updated systematic review and meta-analysis of randomized controlled trials.

OBJECTIVE: The optimal time to start renal replacement therapy (RRT) for acute kidney injury (AKI) remains controversial. We aim to compare the effects of early vs. delayed RRT initiation on clinical outcomes in adult patients with AKI. MATERIALS AND METHODS: PubMed, Embase, Cochrane Library, Web of Science, Chinese Biomedical Literature Database, ClinicalTrials.gov, and the International Clinical Trial registry platform were systematically searched from inception to 7 August 2022. The review included randomized clinical trials (RCTs) comparing early and delayed initiation of RRT in AKI patients. The selected primary outcomes were short-term and long-term mortality. Secondary outcomes included RRT dependency, intensive care unit (ICU) length of stay, hospital length of stay, mechanical ventilator-free days, vasoactive agents-free days, RRT-free days, and adverse events. RESULTS: Overall, 15 RCTs, including 5,625 patients, were analyzed. Early RRT showed no survival benefit when compared to the delayed therapy (28-or 30-day mortality: RR, 1.01, 95% CI: 0.94-1.08, p = 0.87; 60-day mortality: RR, 0.87, 95% CI: 0.71-1.06, p = 0.16; 90-day mortality: RR, 1.00, 95% CI: 0.88-1.13, p = 0.97; in-hospital mortality: RR, 1.05, 95% CI: 0.88-1.24, p = 0.58; ICU mortality: RR, 1.00, 95% CI: 0.91-1.10, p = 0.98). The delayed RRT did not lead to a higher risk of RRT dependency, ICU, or hospital length of stay than the early RRT. Similarly, early initiation of RRT did not lead to longer ventilator-free, vasoactive agent-free, and RRT-free days. However, early RRT initiation was associated with more adverse events. CONCLUSIONS: Our study suggested that early RRT initiation was not associated with survival benefits or better clinical outcomes and increased the risk of RRT-associated adverse events. Current evidence does not support the use of early RRT for AKI patients without urgent indications.

Polyamines conjugated to deoxyribonucleic acid-protein in cell nucleus from filling grain embryos were involved in tolerance of wheat to drought.

Deoxyribonucleic acid-protein (DNAP) of the cell nucleus was purified from developing wheat (Triticum aestivum L.) embryo cells under drought stress, with two cultivars differing in drought tolerance as experimental materials - Longmai No. 079 (drought-tolerant) and Wanmai No. 52 (drought-sensitive). Levels of polyamines (PAs) non-covalently conjugated to the DNA and covalently conjugated to the proteins of DNAP were detected. After soil drought treatment for 10 days, in drought-tolerant Longmai No. 079, the increases in the levels of spermine and spermidine non-covalently conjugated to DNA of DNAP were more statistically significant (P<0.05) than in drought-sensitive Wanmai No. 52. Treatment of Wanmai No. 52 with exogenous Spm could not only enhance the tolerance of the cultivar to drought stress, as judged by flag leaf water content, plasma membrane permeability and grain growth, but also elevate the levels of spermine and spermidine noncovalently conjugated to the DNA of the cultivar. On the contrary, treatment of Longmai No. 079 with methylglyoxyl-bis guanylhydrazone, an inhibitor of S-adenosylmethionine decarboxylase, could significantly (P<0.05) aggravate the drought stress to this cultivar, accompanied by a marked decreases in the levels of spermine and spermidine non-covalently conjugated to the DNA of the cultivar. On the other hand, the content of putrescine covalently conjugated to the proteins of DNAP rose more markedly (P<0.05) in Longmai No. 079 than in Wanmai No. 52. The transglutaminase inhibitor, o-phenanthrolin, could markedly reduce the drought-induced increase in the level of putrescine covalently conjugated to the proteins of DNAP and aggravate drought stress to the two cultivars. Collectively, it could be inferred that spermine and spermidine non-covalently conjugated to the DNA and putrescine covalently conjugated to the proteins of DNAP in the developing grain embryo cell nucleus might enhance the tolerance of wheat plants to soil drought.

Relationship between polyamines conjugated to mitochondrion membrane and mitochondrion conformation from developing wheat embryos under drought stress.

The mitochondrion conformation and the contents of conjugated polyamines were investigated using the embryos of developing wheat (Triticum aestivum L.) grains of two cultivars differing in drought tolerance as experiment materials. After drought stress treatment for 7 days, the relative water content of embryo and relative increase rate of embryo dry weight of the drought-sensitive Yangmai No. 9 cv. decreased more significantly than those of the drought-tolerant Yumai No. 18 cv. Furthermore, the changes in mitochondrion conformation of Yangmai No. 9 were more marked. Meanwhile, the increases of the contents of conjugated non-covalently spermidine (CNC-Spd) and conjugated covalently putrescine (CC-Put) of Yumai No. 18 were more obvious than those of Yangmai No. 9. Treatment with exogenous Spd not only alleviated the injury of drought stress to Yangmai No. 9, but also enhanced the increase of CNC-Spd content and inhibited the change in the mitochondrion conformation of this cultivar. The treatments of Yumai No. 18 with two inhibitors, methylglyoxyl-bis (guanylhydrazone) and phenanthrolin, significantly inhibited the drought stress-induced increases of CNC-Spd and CC-Put contents of the cultivar, respectively. Meanwhile, the treatments with the two inhibitors aggravated the injury of drought stress to Yumai No. 18 and enhanced the change in the mitochondrion conformation of this cultivar. These results mentioned above suggested that the CNC-Spd and CC-Put in embryo mitochondrion membrane isolated from developing grains could enhance the wheat tolerance to drought stress by maintaining the mitochondrion conformation.

Involvement of putrescine in osmotic stress-induced ABA signaling in leaves of wheat seedlings.

To elucidate one mechanism by which putrescine (Put) functions in plant signaling under osmotic stress, Put and ABA contents, and plasma membrane-NADPH oxidase (PM-NOX) activity were detected in wheat seedling leaves. Under osmotic stress, ABA and Put contents, PM-NOX activity, and PM-NOX-dependent O2.- production all increased. The inhibitor tungstate (T) of ABA bio-synthesis reduced the increases in ABA and Put contents under osmotic stress. The inhibitor D-arginine (D-Arg) of Put bio-synthesis didn't reduce osmotic-induced increase of ABA, but it inhibited the increases of PM-NOX activity and O2 . - production, and the inhibitory effects were reversed by exogenous Put. These findings suggested that ABA might regulate Put biosynthesis, and Put might regulate PM-NOX activity. Treatments with three inhibitors imidazole (I), diphenylene iodonium (DPI) and pyridine (P) of PM-NOX reduced significantly not only O2 . - production, but also the stress-induced increase of Put content, which indicated that O2 . - production might regulate Put biosynthesis. Treatments with EGTA (Ca2+ chelator), La3+ and verapamil (V) (Ca2+ channel blockers) reduced significantly the stress-induced increase of Put content, which suggested that Ca2+ might regulate Put biosynthesis. With these findings, it could be concluded that Put was involved in ABA signaling induced by osmotic stress via regulating PM-NOX activity in wheat seedling leaves.

MiR-1271 as a tumor suppressor in breast cancer proliferation and progression via targeting SPIN1.

OBJECTIVE: As breast cancer has become the most common malignant tumor in women worldwide and several microRNAs involved in the mechanism of breast cancer development and progression have been identified, we aimed at investigating the role of miR-1271 in breast cancer. PATIENTS AND METHODS: By quantitative Real-time polymerase chain reaction (qRT-PCR), miR-1271 expression levels in 94 pairs of breast cancer tissue samples and five breast cancer-derived cell lines were detected. Using miR-1271 mimics and inhibitors, the effects of miR-1271 over-expression and knockdown on the proliferation, invasion and migration of MCF-7 cells were analyzed, respectively. Dual-luciferase activity assay was recruited to examine the potential target gene SPIN1 that was predicted by several databases. Protein level was studied using Western blotting. RESULTS: MiR-1271 was significantly lowly expressed in breast cancer tissue samples and cell lines. Over-expression of miR-1271 in MCF-7 cells significantly decreased the cell proliferation, invasion, and migration abilities while down-regulation of miR-1271 in MDA-MB-453 cells increased these abilities oppositely. Dual-luciferase and Western blotting were used to confirm SPIN1 as a target gene of miR-1271. Furthermore, up-regulation SPIN1 reserved the suppressive effect of miR-1271 over-expression on cell growth and progression. CONCLUSIONS: miR-1271 can suppress breast cancer cell proliferation and progression via SPIN1, which may provide a potential therapeutic target in treatment for breast cancer.

Photoactivation of hypericin down-regulates glutathione S-transferase activity in nasopharyngeal cancer cells.

Photodynamic therapy (PDT) is a new modality of treatment for cancer. Hypericin is a photosensitizer, which is known to generate reactive oxygen species upon activation with light. We observed that photoactivated hypericin induces the generation of reactive oxygen intermediates in nasopharyngeal cancer (NPC) cells in vitro. There was also significant reduction of Glutathione S-transferase (GST) activity in HK1 and CNE-2 NPC cells and in tumor tissues from the NPC/HK1 murine tumor model by hypericin-mediated PDT. As antioxidants protect cells against phototoxicity, down-regulation of GST activity would potentiate the efficacy of hypericin-PDT treatment.

[Correlation between the age-related memory impairment and the level of free Ca(2+) in brain synaptosomes].

In this study, We investigated the changes in the ability of learning-memory of 12-month- and 18-month-old mice, and the intrasynaptosomal Ca(2+) levels of some mouse brain areas (hippocampus, cerebral cortex, corpus quadrigem and cerebellum) on the basis of our last behavior observation and biochemistry detection. Meanwhile, the intrasynaptosomal Ca(2+) levels of the four brain areas were compared between the mice of impaired and good memory. The main results are as follows. With the increase of age, the ability of learning-memory of mice decreased significantly; and the concentration of the intrasynaptosomal free Ca(2+) (i) in mouse brain areas (except for cerebral cortex) increased markedly, especially in the memory-impaired mice. The results suggest that age-related memory impairment may be associated with the overloading of intrasynaptosomal free Ca(2+) (i).

[Correlation between ability of learning-memory and synaptosomal free [Ca2+]i in mice of different age].

In the present investigation, the behavior of learning and memory of 1-month and 6-month-old mice was studied by using Y-maze and one-trial passive avoidance response device. The synaptosomal free [Ca2+]i of four main brain regions (Hippocampus, Cerebral cortex, Cerebellum, Tectum of midbrain) of these mice were measured by fluorescent probe Ca2+ indicator Fura-2 and an AR-CM-MIC cation measurement system. The results showed that, in comparison with 1-month-old mice, the ability of discrimination learning and memory of 6-month-old ones were attenuated, and the synaptosomal free [Ca2+]i of hippocampus was increased.

[Regulation of NGF on intra-synaptosomal free [Ca2+]i in mouse brain].

In the present study, the influence of direct intrahippocampal injection of NGF (nerve growth factor) on synaptosomal free [Ca2+]i in 1-month-old and 18-month-old mice was observed. The regulation of NGF on synaptosomal free [Ca2+]i in vitro was also investigated. The synaptosomal free [Ca2+]i in the main brain regions (cerebral cortex, hippocampus, cerebellum, diencephalon) of these mice was measured by fluorescent probe Fura-2 of Ca2+ indicator and an AR-CM-MIC cation measurement system. The results are as follows: (1) NGF showed no detectable effect on hippocampal intrasynaptosomal free [Ca2+]i in 1-month-old mice, but appropriate dose of NGF was shown to markedly reduce [Ca2+]i in 18-month-old mice. (2) Under the condition of low [Ca2+]i level caused by the drug, NGF was found to increase [Ca2+]i; conversely, was reduced [Ca2+]i when intracellular Ca2+ was overloaded. These results suggest that NGF has dual regulation on Ca2+ level in brain and this might be an important mechanism for its action in improving memory lesion, in aged person.

Hypericin photoactivation triggers down-regulation of matrix metalloproteinase-9 expression in well-differentiated human nasopharyngeal cancer cells.

Recently, we have shown that hypericin-mediated photodynamic therapy (PDT) is a promising modality for the treatment of nasopharyngeal cancer (NPC). The present study evaluated the expression of matrix metalloproteinase-9 (MMP-9) following hypericin-PDT in well-differentiated HK1 NPC cells. Down-regulation of MMP-9 by hypericin-PDT was observed at the mRNA level in HK1 cells in vitro and in vivo and at the protein level in vitro. Transcriptional activities of the activator protein-1 (AP-1) and nuclear factor (NF)-kappaB regulatory elements were inhibited by PDT. We also found that PDT reduced secreted granulocyte-macrophage colony stimulating factor (GM-CSF), which is known to activate transcription of NK-kappaB and AP-1. However, incubation of untreated HK1 cells with exogenous GM-CSF abrogated the reduction of MMP-9 production in hypericin-PDT-treated cells. It would appear that PDT downregulates MMP-9 expression via inhibition of GM-CSF production, which in turn modulates AP1/NF-kappaB transcriptional activities. Suppression of MMP-9 by hypericin-PDT may have therapeutic implications.