The use of melittin to enhance transgene expression mediated by recombinant adeno-associated virus serotype 2 vectors both in vitro and in vivo.

OBJECTIVE: Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus (rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based rAAV2 vector delivery strategy. METHODS: The melittin peptide was inserted into the rAAV2 capsid either in the loop VIII of all viral proteins (VPs) or at the N terminus of VP2. Various rAAV2-gfp or -fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression. RESULTS: rAAV2 vectors with melittin peptide inserted in the loop VIII of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of rAAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity. CONCLUSION: Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced rAAV2-mediated transgene expression. Although further in vivo evaluations are required, this research not only expands the pharmacological potential of melittin, but also provides a new strategy to improve gene therapy mediated by rAAV vectors.

Acupuncture for glaucoma-induced optic atrophy: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the clinical effect of acupuncture for glaucoma-induced optic atrophy.@*METHODS@#A total of 70 patients (89 affected eyes) with glaucoma-induced optic atrophy were randomized into an observation group and a control group, 35 cases in each group. The control group was given basic western medicine treatment. In the observation group, on the basis of the treatment in the control group, acupuncture was applied at main acupoints i.e. Baihui (GV 20), Shangjingming (Extra), Chengqi (ST 1), Fengchi (GB 20), Zusanli (ST 36), combined with supplementary acupoints based on syndrome differentiation, once every three days, twice a week. The treatment for 3 months was required in both groups. Before treatment, after treatment and in follow-up of 6 months after treatment, the best corrected visual acuity (BCVA), intraocular pressure (IOP), indexes of visual field (visual field index [VFI], mean deviation [MD], pattern standard deviation [PSD]) and mean thickness of retinal nerve fiber layer (RNFL) were observed in the two groups.@*RESULTS@#Compared before treatment, BCVA was decreased after treatment and in follow-up in the control group (P<0.05); in the follow-up, BCVA in the observation group was higher than that in the control group (P<0.05). On each time point before and after treatment, there was no significant difference within or between the two groups (P>0.05). After treatment and in the follow-up, the mean thickness of RNFL was larger than the control group (P<0.05).@*CONCLUSION@#On the basis of the basic western medicine treatment, acupuncture can delay the decline of vision and the thinning of retinal nerve fiber layer in patients with glaucoma-induced optic atrophy.

Effect of Bushen Chushi() decoction combined with platelet-rich plasma on regulating TGF-β1 and Smad-1 expression in serum of knee osteoarthritis patient / 中国骨伤

OBJECTIVE@#To investigate the effect of Bushen Chushi decoction combined with platelet-rich plasma(PRP) to treat knee osteoarthritis(KOA) in early and middle stage and its regulation on TGF-β1 and Smad-1 expression in serum.@*METHODS@#Total of 45 patients with KOA in early and middle stage from May 2020 to April 2022 were treated and divided into control group and observation group. In control group, there were 30 patients including 12 males and 18 females, aged from 43 to 69 years old with an average of(57.3±6.5) years old and disease duration ranged from 1.5 to 5.0 years with an average of(3.8±1.7) years, and there were 8 cases in gradeⅠ, 13 cases in gradeⅡ, and 9 cases in grade Ⅲ according to Kellgren-Lawrence Grade, PRP 5 ml was injected into knee joint on the first day of No1, 3 week together for 2 times. In the observation group, there were 15 cases including 7 males and 8 females, aged from 45 to 70 years old with an average of (56.7±6.2) years old and disease duration ranged from 1.8 to 5.7 years with an average of (4.0±1.8) years, there were 4 cases in gradeⅠ, 9 cases in gradeⅡand 4 cases in grade Ⅲ according to the Kellgren-Lawrence Grade, PRP 5 ml were injected into knee joints that the time and frequency were the same as those in the control group, and at the same time Bushen Chushi decoction orally were taken 1 dose per day with a total of 28 doses. All patients were treated for four weeks. Visual analogue scale(VAS) and Lequesne MG score before and after treatment were used to evaluate improvement of knee pain and joint function. The TGF-β1 and Smad-1 levels in serum were measured before and after treatment in two groups. The incidence of complications in two groups was observed.@*RESULTS@#All patients were followed up for 26 to 30 days with an average of (28.0±0.6) days. There was no significant difference in VAS and knee Lequesne MG scores between two groups before treatment(P>0.05). The scores of VAS and knee Lequesne MG on the first day after treatment in both groups were lower than those before treatment(P<0.05). The VAS and knee Lequesne MG scores in observation group were lower than those in control group(P<0.05) on the first day after treatment. The TGF-β1 level in serum after treatment were higher significantly than that before treatment in two groups(P<0.05). After treatment, TGF-β1 level in serum in observation group were lower than those in control group with statistically significant differences(P<0.05). The Smad-1 levels in serum after treatment in observation group were higher significantly than that in control group(P<0.05). The levels of Smad-1 were not statistically significant between before and after treatment(P>0.05). There was no significant difference in postopertaive complications between two groups (P>0.05).@*CONCLUSION@#The efficacy of Bushen Chushi decoction combined with PRP in treatment of early and middle KOA is better than that of PRP injection alone. The combined treatment could reduce TGF-β1 level and increase Smad-1 level in serum, which may be a mechanism to inhibit inflammation and alleviate cartilage degeneration to some extent.

The Moderating Effect of Compassionate Mindfulness on the Psychological Needs and Emotions of Generation Y in the 21st Century in Taiwan.

During the 2020 COVID-19 pandemic in Taiwan, 6.5% of Generation Y required medical treatment for emotional and stress-related mental disorders. This study explores the moderating effect of mindfulness training on psychological needs and emotions to propose effective measures to promote the mental health of Generation Y. This study was carried out by questionnaire, using the data of respondents born in 1980-1999, collected in three different periods for quantitative analysis with compassionate mindfulness as the main variable. The results show that the compassionate mindfulness effect on emotion regulation varies greatly among different educational levels. However, it still plays a positive role in the psychological needs of Generation Y. Most members of Generation Y who receive compassionate mindfulness training have fewer basic needs and more interpersonal trust. They pay more attention to individual-oriented self-realization. Compassionate mindfulness has a greater positive moderating effect on the mental health of women aged 30-39 and those who are highly educated. Compassionate mindfulness has a more positive moderating effect on the psychological needs of members of Generation Y who were born more recently. During the COVID-19 pandemic, providing compassionate mindfulness has a significant positive effect on the prevention of mental disorders of Generation Y in Taiwan.

[Icariin improves renal interstitial fibrosis in a rat model of chronic renal failure by regulating mitochondrial dynamics].

This study aims to explore the effect of icariin(ICA) on mitochondrial dynamics in a rat model of chronic renal failure(CRF) and to investigate the molecular mechanism of ICA against renal interstitial fibrosis(RIF). CRF was induced in male Sprague-Dawley(SD) rats with 5/6(ablation and infarction, A/I) surgery(right kidney ablation and 2/3 infarction of the left kidney). Four weeks after surgery, the model rats were randomized into the following groups: 5/6(A/I) group, 5/6(A/I)+low-dose ICA group, and 5/6(A/I)+high-dose ICA group. Another 12 rats that received sham operation were randomly classified into 2 groups: sham group and sham+ICAH group. Eight weeks after treatment, the expression of collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), mitochondrial dynamics-related proteins(p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2), and mitochondrial function-related proteins(TFAM, ATP6) in the remnant kidney tissues was detected by Western blot. The expression of α-smooth muscle actin(α-SMA) was examined by immunohistochemical(IHC) staining. The NRK-52 E cells, a rat proximal renal tubular epithelial cell line, were cultured in vitro and treated with ICA of different concentration. Cell viability was detected by CCK-8 assay. In NRK-52 E cells stimulated with 20 ng·mL~(-1) TGF-ß1 for 24 h, the effect of ICA on fibronectin(Fn), connective tissue growth factor(CTGF), p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 was detected by Western blot, and the ATP content and the mitochondrial morphology were determined. The 20 ng·mL~(-1) TGF-ß1-stimulated NRK-52 E cells were treated with or without 5 µmol·L~(-1) ICA+10 µmol·L~(-1) mitochondrial fusion promoter M1(MFP-M1) for 24 h and the expression of fibrosis markers Fn and CTGF was detected by Western blot. Western blot result showed that the levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were increased and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were decreased in 5/6(A/I) group compared with those in the sham group. The levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were significantly lower and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were significantly higher in ICA groups than that in 5/6(A/I) group. IHC staining demonstrated that for the expression of α-SMA in the renal interstitium was higher in the 5/6(A/I) group than in the sham group and that the expression in the ICA groups was significantly lower than that in the 5/6(A/I) group. Furthermore, the improvement in the fibrosis, mitochondrial dynamics, and mitochondrial function were particularly prominent in rats receiving the high dose of ICA. The in vitro experiment revealed that ICA dose-dependently inhibited the increase of Fn, CTGF, and p-Drp1 S616, increased p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6, elevated ATP content, and improved mitochondrial morphology of NRK-52 E cells stimulated by TGF-ß1. ICA combined with MFP-M1 further down-regulated the expression of Fn and CTGF in NRK-52 E cells stimulated by TGF-ß1 compared with ICA alone. In conclusion, ICA attenuated RIF of CRF by improving mitochondrial dynamics.

[Electroacupuncture of "Shangjuxu"(ST37) and "Tianshu"(ST25) reduces colonic injury by suppressing NF-κB/NLRP3 signaling in rats with ulcerative colitis].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Shangjuxu"(ST37) and "Tianshu"(ST25) on colonic mucosal injury and activities of nuclear factor-κB (NF-κB)/Nod-like receptor family，pyrin domain-containing 3 (NLRP3) inflammasome signaling in the colonic tissue in rats with ulcerative colitis (UC), so as to explore its mechanisms underlying improvement of UC. METHODS: Male SD rats were randomly divided into blank, model, medication and EA groups, with 12 rats in each group. The UC model was established by enema of 2-4-6 trinitrobenzene sulfonic acid +50% ethanol (2.5 mL). EA (10 Hz/50 Hz) was applied to bilateral ST37 and ST25 for 20 min, once a day, for a total of 10 days. Rats of the medication group received gavage of mesalazine suspension (2 mL：0.2 g/kg+0.9% saline) once a day, for 10 days. The rats' general conditions were recorded for calculating the disease activity index (DAI) score (0-4 points). The colonic tissue was sampled for giving colonic mucosa damage index (CMDI, 0-5 points) score and for observing histopathological changes after hematoxylin-eosin (HE) staining, and for detecting expression levels of NF-κB and NLRP3 by using immunohistochemistry and Western blot, separately. The contents of serum interleukin-1ß (IL-1ß), NLRP3 and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay. RESULTS: Compared with the blank group, the DAI and CMDI scores, contents of serum IL-1ß, NLRP3, and TNF-α, as well as the immunoactivity and expression of NF-κB and NLRP3 proteins were significantly increased in the model group (P<0.05). Relevant to the model group, modeling-induced increases of DAI and CMDI scores, serum IL-1ß, NLRP3 and TNF-α contents, and NF-κB and NLRP3 expression were reversed in both medication and EA groups (P<0.05), the effect of EA was apparently superior to that of mesalazine in down-regulating CMDI score and serum IL-1ß level (P<0.05). No significant diffe-rences were found between the medication and EA groups in down-regulating DAI score, serum TNF-α and NLRP3 contents, and expression of NF-κB and NLRP3 proteins (P>0.05). The rats' general conditions including arch back sloth, anorexia, loss of fur gloss, weight loss, lethargy and loose of stool, and histopathological changes such as necrosis of intestinal mucosa, formation of obvious ulcerative surface, with many neutrophils and pus cells and inflammatory cell infiltration were obvious in the model group, which were relative milder in both medication and EA groups. CONCLUSION: EA can relieve colonic injury in UC rats, which may be related to its functions in down-regulating serum IL-1ß, TNF-α and NLRP3 levels by suppressing colonic NF-κB / NLRP3 inflammasome signaling.

Green tea powder inclusion promoted hatchability through increased yolk antioxidant activity.

Dietary supplementation of green tea powder (GTP) changes egg quality of hens, however, whether these changes affect incubation is still unknown. This study was to compare the proteomic difference of incubated eggs from hens with GTP supplemented or not. Huainan partridge chickens (1,080) at 35 wk of age were allocated into 2 groups, one group fed basal diet (CG) and one group fed basal diet plus 1% GTP (EG). After 4 wk feeding, artificially fertilized eggs were collected for yolk cholesterol determination and incubation. During incubation, 6 embryos from each group were randomly selected in each day for yolk protein extraction and quantification. Yolk cholesterol content was significantly lower, while the hatchability was significantly higher in EG than that of the CG group (P < 0.05). Yolk protein concentration at embryonic days (ED) of 0, 2, 6, and 13 showed significant changes and were selected for proteomic analysis by 2-dimensional gel electrophoresis combined with liquid chromatography-tandem mass spectrometry. Fifty-one differentially expressed (DE) protein spots were identified among different incubation stages between CG and EG group which were mainly classified into vitellogenin, immunoglobulin, and ovoinhibitor, and occupied 45.1, 23.5, and 15.7%, respectively, to the total DE proteins. Ovotransferrin, participated in extracellular sequestering of iron ion process, was significantly lower in EG group than that of the CG group (P < 0.05). Ig light chain precursor (Immunoglobulin) exhibited higher expression at ED6 in EG group as compared with that of the CG group, and was participated in immune response related processes. Ovoinhibitor, mainly involved in protease binding activity, showed lower abundance at ED13 in EG group as compared with that of the CG group. Vitellogenin-3, showed lower expression in EG group as compared with that of the CG group, was mainly participated in lipid transportation and localization according to GO enrichment. Chickens fed diet with GTP provided eggs more antioxidant ability that increased hatchability, indicated that GTP could be considered as additive in breeding layer.

Efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo and its effect on skin sympathetic response / 中国基层医药

Objective:To investigate the efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo (BPPV) and its effect on skin sympathetic response.Methods:A total of 110 patients with residual dizziness after successful reduction of BPPV who were admitted to Wenzhou Hospital of Traditional Chinese Medicine from January 2019 to May 2021 were included in this study. They were randomly divided into control and study groups, with 55 patients in each group. Patients in the control group were treated with drugs, and those in the study group were treated with auricular acupoint bean-embedding therapy. Before and after treatment, dizziness handicap inventory (DHI) score, activities-specific balance confidence (ABC) score, and Hamilton Anxiety Scale (HAMA) score were compared between the two groups. The latency and amplitude of sympathetic skin response (SSR) were measured.Results:After 15 days of treatment, DHI scores of all dimensions in the study group were significantly lower than those in the control group ( t = 16.13-20.62, all P < 0.05). ABC score in the study group was significantly higher than that in the control group [(87.90 ± 6.01) points vs. (80.55 ± 8.73) points, t = 3.10, P < 0.05). HAMA score in the study group was significantly lower than that in the control group [(7.85 ± 1.06) points vs. (13.30 ± 2.49) points, t = 8.43, P < 0.001]. The initial latency value of SSR in the study group was significantly higher than that in the control group [(1.95 ± 0.27) ms vs. (1.67 ± 0.21) ms, and the amplitude of SSR in the study group was significantly lower than that in the control group [(1.49 ± 0.15) mV vs. (1.70 ± 0.22) mV, t = 4.73, 4.04, both P < 0.001]. The incidence of adverse reactions in the study group was significantly lower than that in the control group [0.0% (0/55) vs. 10.9% (6/55), χ2 = 4.40, P < 0.05]. Conclusion:Auricular acupoint bean-embedding therapy can effectively alleviate the symptoms of dizziness after successful reduction of BPPV, improve patient's psychological status and autonomic nerve function, which is worthy of clinical promotion.

Application of acupuncture in inhibiting intestinal peristalsis in colonoscopy / 中国针灸

Acupuncture regulating gastrointestinal motility has the characteristics of bidirectional benign regulation, acupoint specificity and immediacy. And its regulation is mainly achieved through the "neuro-endocrine-immune" network system. Acupuncture at Neiguan (PC 6) and Hegu (LI 4) to inhibit intestinal peristalsis may have good application value in colonoscopy.

Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo preliminarily predict the active components， action targets， and signaling pathways of Arnebia euchroma in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in in vitro experiments. MethodThe active components and related targets of A. euchroma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of A. euchroma and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of A. euchroma against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in A. euchroma， was docked to the target by AutoDock Vina 1.1.2. The in vitro experiments were then carried out to verify the anti-melanoma effect of A. euchroma. ResultA total of 271 common targets of A. euchroma and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis， A. euchroma mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （P<0.05）. ConclusionA. euchroma alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with A. euchroma.

Identification of chemical constituents of Xiaochengqi Decoction by UPLC-Q-TOF/Fast DDA combined with UNIFI software / 中国中药杂志

Based on the combination of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF) and Waters UNIFI software, the chemical constituents of the classic prescription Xiaochengqi Decoction were qualitatively analyzed and identified. The UPLC conditions are as follows: Acquity HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm), column temperature of 30 ℃, mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B), and flow rate of 0.3 mL·min~(-1). High-resolution MS data of Xiaochengqi Decoction were collected in ESI~(+/-) modes by Fast DDA. The structures of the chemical constituents were tentatively characterized or identified by UNIFI software according to the retention time of reference standards and characteristic fragment ions in MS profile, and literature data. A total of 233 components in Xiaochengqi Decoction were identified, with 93 from wine-processed Rhei Radix et Rhizoma, 104 from bran-processed Aurantii Fructus Immaturus, and 36 from ginger-processed Magnoliae Officinalis Cortex. These 233 components included anthraquinones, flavonoids, lignans, alkaloids, coumarins, and phenylethanoid glycosides. The result provided experimental evidence for the further study on establishment of quality standard and product development of the formula.

Icariin improves renal interstitial fibrosis in a rat model of chronic renal failure by regulating mitochondrial dynamics / 中国中药杂志

This study aims to explore the effect of icariin(ICA) on mitochondrial dynamics in a rat model of chronic renal failure(CRF) and to investigate the molecular mechanism of ICA against renal interstitial fibrosis(RIF). CRF was induced in male Sprague-Dawley(SD) rats with 5/6(ablation and infarction, A/I) surgery(right kidney ablation and 2/3 infarction of the left kidney). Four weeks after surgery, the model rats were randomized into the following groups: 5/6(A/I) group, 5/6(A/I)+low-dose ICA group, and 5/6(A/I)+high-dose ICA group. Another 12 rats that received sham operation were randomly classified into 2 groups: sham group and sham+ICAH group. Eight weeks after treatment, the expression of collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), mitochondrial dynamics-related proteins(p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2), and mitochondrial function-related proteins(TFAM, ATP6) in the remnant kidney tissues was detected by Western blot. The expression of α-smooth muscle actin(α-SMA) was examined by immunohistochemical(IHC) staining. The NRK-52 E cells, a rat proximal renal tubular epithelial cell line, were cultured in vitro and treated with ICA of different concentration. Cell viability was detected by CCK-8 assay. In NRK-52 E cells stimulated with 20 ng·mL~(-1) TGF-β1 for 24 h, the effect of ICA on fibronectin(Fn), connective tissue growth factor(CTGF), p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 was detected by Western blot, and the ATP content and the mitochondrial morphology were determined. The 20 ng·mL~(-1) TGF-β1-stimulated NRK-52 E cells were treated with or without 5 μmol·L~(-1) ICA+10 μmol·L~(-1) mitochondrial fusion promoter M1(MFP-M1) for 24 h and the expression of fibrosis markers Fn and CTGF was detected by Western blot. Western blot result showed that the levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were increased and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were decreased in 5/6(A/I) group compared with those in the sham group. The levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were significantly lower and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were significantly higher in ICA groups than that in 5/6(A/I) group. IHC staining demonstrated that for the expression of α-SMA in the renal interstitium was higher in the 5/6(A/I) group than in the sham group and that the expression in the ICA groups was significantly lower than that in the 5/6(A/I) group. Furthermore, the improvement in the fibrosis, mitochondrial dynamics, and mitochondrial function were particularly prominent in rats receiving the high dose of ICA. The in vitro experiment revealed that ICA dose-dependently inhibited the increase of Fn, CTGF, and p-Drp1 S616, increased p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6, elevated ATP content, and improved mitochondrial morphology of NRK-52 E cells stimulated by TGF-β1. ICA combined with MFP-M1 further down-regulated the expression of Fn and CTGF in NRK-52 E cells stimulated by TGF-β1 compared with ICA alone. In conclusion, ICA attenuated RIF of CRF by improving mitochondrial dynamics.

Analysis of muscle activation in children and adolescents with severe cerebral palsy / Análise da ativação muscular em crianças e adolescentes com paralisia cerebral grave

Abstract Introduction: Children and adolescents with spastic quadriparesis have a worse selective motor control, and studies with this population are still very scarce. The same is true with scientific evidence of one of the methods most used as a physiotherapeutic treatment in this population, the Bobath Concept. Objective: To evaluate spine erector muscles activation, gluteus medius and gluteus maximus, through the handling of the Bobath Concept and the sustained kneeling posture in subjects with a diagnosis of severe cerebral palsy; and to compare muscle activation with a reference group, in order to increase the reliability of this study. Methods: A cross-sectional study was carried out with 38 children and adolescents with cerebral palsy, classified by GMFCS at levels IV and V, and 20 healthy participants, aged between 3 and 18 years. They were submitted to the handling of the Bobath Concept and to the sustained kneeling posture, with muscle activation obtained by electromyography. Results: We observed significant muscle activation during handling in side-sitting, with weight transfer and without the help of another therapist, and in the sustained kneeling posture, for the erector of the spine and gluteus medius. Conclusion: The evidence from this study suggests that both the handling in side-sitting and the sustained kneeling posture cause significant muscle activation in the erector of the spine and gluteus medius for severe quadriparesis subjects, GMFCS IV and V, which can contribute to the improvement of postural control and decision-making in physical therapy practice.

Resumo Introdução: Crianças e adolescentes com quadriparesia espástica apresentam pior controle motor seletivo e estudos com essa população ainda são muito escassos. O mesmo ocorre com as evidências científicas de um dos métodos mais utilizados como tratamento fisioterapêutico nessa população, o Conceito Bobath. Objetivo: Avaliar a ativação dos músculos eretores da coluna, glúteo médio e glúteo máximo por meio do manuseio do Conceito Bobath e da postura ajoelhada sustentada, em indivíduos com diagnóstico de paralisia cerebral grave; e comparar a ativação muscular com um grupo de referência, a fim de aumentar a confiabilidade deste estudo. Métodos: Realizou-se um estudo transversal com 38 crianças e adolescentes com paralisia cerebral, classificados pelo GMFCS nos níveis IV e V, e 20 participantes saudáveis com idade entre 3 e 18 anos. Eles foram submetidos ao manuseio do Conceito Bobath e à postura ajoelhada sustentada, com ativação muscular obtida por eletromiografia. Resultados: Observou-se ativação muscular significativa durante o manuseio na posição sentada de lado, com transferência de peso e sem auxílio de outro terapeuta, e na postura ajoelhada sustentada para o eretor da coluna e glúteo médio. Conclusão: As evidências deste estudo sugerem que tanto o manuseio na posição sentada de lado quanto a sustentação da postura ajoelhada causam significativa ativação muscular no eretor da coluna e glúteo médio para indivíduos com quadriparesia grave, GMFCS IV e V, o que pode contribuir para a melhora do controle postural e tomada de decisão na prática fisioterapêutica.

The use of miR122 and its target sequence in adeno-associated virus-mediated trichosanthin gene therapy.

OBJECTIVE: Plant-derived cytotoxic transgene expression, such as trichosanthin (tcs), regulated by recombinant adeno-associated virus (rAAV) vector is a promising cancer gene therapy. However, the cytotoxic transgene can hamper the vector production in the rAAV producer cell line, human embryonic kidney (HEK293) cells. Here, we explored microRNA-122 (miR122) and its target sequence to limit the expression of the cytotoxic gene in the rAAV producer cells. METHODS: A miR122 target (122T) sequence was incorporated into the 3' untranslated region of the tcs cDNA sequence. The firefly luciferase (fluc) transgene was used as an appropriate control. Cell line HEK293-mir122 was generated by the lentiviral vector-mediated genome integration of the mir122 gene in parental HEK293 cells. The effects of miR122 overexpression on cell growth, transgene expression, and rAAV production were determined. RESULTS: The presence of 122T sequence significantly reduced transgene expression in the miR122-enriched Huh7 cell line (in vitro), fresh human hepatocytes (ex vivo), and mouse liver (in vivo). Also, the normal liver physiology was unaffected by delivery of 122T sequence by rAAV vectors. Compared with the parental cells, the miR122-overexpressing HEK293-mir122 cell line showed similar cell growth rate and expression of transgene without 122T, as well as the ability to produce liver-targeting rAAV vectors. Fascinatingly, the yield of rAAV vectors carrying the tcs-122T gene was increased by 77.7-fold in HEK293-mir122 cells. Moreover, the tcs-122T-containing rAAV vectors significantly reduced the proliferation of hepatocellular carcinoma cells without affecting the normal liver cells. CONCLUSION: HEK293-mir122 cells along with the 122T sequence provide a potential tool to attenuate the cytotoxic transgene expression, such as tcs, during rAAV vector production.

Study on the role of cupping therapy on the implementation of the constitution nursing of elderly hypertension patients with phlegm-dampness constitution in the community / 中国实用护理杂志

Objective:To explore the effect of cupping therapy on regulating phlegm dampness constitution of elderly patients with hypertension in the community.Methods:From August to November in 2019, 120 elderly patients with phlegm dampness hypertension were randomly divided into control group and intervention group, 60 cases in each. The control group was given traditional Chinese medicine (TCM) health care in the community, and the intervention group was given cupping therapy intervention on this basis, to evaluate the changes of the scores of TCM phlegm wet body mass table, the total score reduction rate of TCM phlegm wet body mass, the conversion rate of TCM phlegm wet body mass, blood pressure, body mass index and waist circumference between the two groups before the intervention, 1 month, 2 months, 3 months after the intervention.Results:Finally, there were 51 cases in the intervention group and 57 cases in the control group. The TCM phlegm dampness constitution sub-scale and total scores of the comparison between groups at four different time points showed that except for the dimension of abdominal hypertrophy, the sub-scale and total scores of TCM phlegm-dampness constitution scale in the two groups were lower than before, and the differences were statistically significant ( F values were 3.450-94.995, P<0.05). The total score reduction rate of phlegm dampness constitution in the intervention group was 0.17 ± 0.10, higher than that in the control group (0.12 ± 0.09), and the difference was statistically significant ( t value was - 2.773, P < 0.05). The transformation rate of phlegm-dampness constitution was 47.1% (24/51) in the intervention group, which was higher than that in the control group (33.3%,19/57). The value of systolic blood pressure was lower in the intervention group [(135.10±2.64) mmHg,1 mmHg=0.133 kPa] than that in the control group [(138.88±2.42) mmHg] after 2 months of intervention, and the difference was statistically significant ( F value was 6.586, P<0.05). The value of diastolic blood pressure was lower in the intervention group [(80.75±3.03) mmHg] than that in the control group [(78.14±3.43) mmHg] after 3 months of intervention, and the difference was statistically significant ( F value was 6.758, P<0.05). Conclusions:On the basis of TCM physical nursing in the community, the effect of cupping therapy on reducing the score of phlegm and dampness physical fitness of elderly hypertension patients in the community, promoting the transformation of phlegm and dampness physical fitness of elderly hypertension patients in the community, and reducing the blood pressure level of elderly hypertension patients in the community is better than that of TCM physical nursing in the community, which is worth being promoted in the community.

Neuroprotective Effect of Fructus broussonetiae on APP/PS1 Mice via Upregulation of AKT/β-Catenin Signaling / 中国结合医学杂志

OBJECTIVE@#To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae (FB) in both mouse and cell models of Alzheimer's disease (AD).@*METHODS@#APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity. RNA-Seq, Western blotting, and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice. To further explore the mechanisms underlying FB's protective effect, PC-12 cells were treated with Aβ@*RESULTS@#FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests. RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways, specifically decreasing cell apoptotic signaling and increasing AKT and β-catenin signaling. Similarly, FB up-regulated both AKT and β-catenin signaling in PC-12 cells pre-treated with Aβ@*CONCLUSIONS@#FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice, as well as improved cell viability in an in vitro model of AD. The protective actions of FB occurred via the upregulation of AKT/β-catenin signaling.

The effect and influencing factors of percutaneous microwave ablation for the treatment of benign thyroid nodules / 中国医师杂志

Objective:To investigate the effect and influencing factors of microwave ablation (MVA) in the treatment of benign thyroid nodules.Methods:The clinical data of ultrasound-guided microwave ablation for thyroid benign nodules in the Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from April 2017 to April 2019 were retrospectively analyzed. At 1, 3 and 6 months after operation, conventional ultrasound examination was performed to calculate the volume reduction rate of the nodules. The nodules were divided into groups according to gender, age, nodule blood supply, nodule size, nodule nature and Hashimoto′s thyroiditis background, and the related factors influencing microwave ablation were analyzed.Results:68 patients (106 nodules) with benign thyroid nodules were treated with microwave ablation. The volume of benign thyroid nodules after the MWA treatment was significantly reduced after 1, 3, 6 months, and their nodule volume reduction ratio (VRR) were (39.7±6.1)% (1 months), (56.2±5.9)% (3 months), (70.3±5.4)% (6 months), respectively. There were significant differences in the volume reduction ratio of nodules at 1, 3 and 6 months after operation among different nodule size, nodule nature and Hashimoto′s thyroiditis background, with statistically significant difference ( P<0.05). However, there was no significant difference in the reduction ratio of nodules in different gender, age and nodule blood supply at 1, 3 and 6 months after operation ( P>0.05). Pearson correlation analysis showed that VRR was negatively correlated with ablation time per unit volume, with statistically significant difference ( P<0.05). Logistic regression analysis indicated that only nodule nature and ablation time per unit volume entered the regression equation. Conclusions:The size and nature of the nodules, Hashimoto′s thyroiditis background and ablation time per unit volume will affect the postoperative volume reduction rate.

Mild iron overload induces TRIP12-mediated degradation of YY1 to trigger hepatic inflammation.

Increasing populations are found to bear mild hepatic iron overload (HIO) due to unhealthy lifestyles, metabolic diseases, etc., whether this mild but chronic HIO induces hepatic inflammation is unknown. In the present study, mice receiving a 12-months 0.3% dextran-iron diet show mild HIO with no detectable oxidative damages in the liver but have infiltrated macrophages and increased IL-6, TNFα, AST and ALT since 6-months. The HNF4α/miR-122/CCL2 pathway, identified by our previous studies to induce macrophages infiltration, is initiated by chronic mild HIO. After excluding the role of DNA methylation, a modified transcription factor microarray is applied to find that transcription factor YY1 is responsible for HIO-decreased HNF4α expression. Then the E3 ubiquitin ligase TRIP12 is identified by an immunoprecipitation coupled LC-MS/MS and proved to bind and ubiquitinate YY1, leading to its degradation. The overexpression or silence of YY1 in the liver regulates the HNF4α/miR-122/CCL2 pathway. More importantly, YY1 overexpression alleviates chronic mild HIO induced hepatic inflammatory responses. In conclusion, these results elucidate an oxidative-stress-independent, TRIP12/YY1/HNF4α/miR-122/CCL2 pathway of chronic mild HIO inducing hepatic inflammation, implying that effective measures in addition to antioxidants are needed for individuals at the risk of chronic mild HIO.

The improved targeting of an aspirin prodrug albumin-based nanosystem for visualizing and inhibiting lung metastasis of breast cancer.

Lung metastasis is the principal reason for the majority of deaths from breast cancer. The nonsteroidal anti-inflammatory drug aspirin can prevent lung metastasis in breast tumors via inhibiting heparanase. However, the lack of specific targets and limited accumulation at the site of the tumor have thus far hindered the use of aspirin in oncotherapy. In this study, we developed the nanoplatform FA-BSA@DA and loaded it with the versatile aspirin prodrug DA to visualize and inhibit breast cancer metastasis via targeting heparanase. This nanosystem can be effectively targeted to folic acid (FA)-positive tumor cells, and would then subsequently release a high dose of DA, whose ester bond is specifically ruptured by H2O2 in the tumor microenvironment to afford the therapeutic drug aspirin and near-infrared (NIR) fluorescent reporter DCM. The released aspirin can effectively prevent breast cancer lung metastasis through the inhibition of heparanase activity, and the NIR fluorescent signals emitted from DCM can be used to monitor and evaluate the metastasis levels of breast cancer. Our results showed that the expression of heparanase was significantly decreased, and lung metastasis from breast cancer was effectively monitored and inhibited after treatment with FA-BSA@DA. Furthermore, the collaborative therapy nanoplatform FA-BSA@DA/DOX exhibited strong therapeutic effects in the treatment of breast cancer in vitro and in vivo via the introduction of doxorubicin (DOX) to the system, which resulted in an even stronger result due to its synergistic effects with aspirin. This heparanase-reliant strategy has profound significance for the extended development of nanoplatforms based on versatile aspirin prodrugs, which may offer a solution to clinically prevent breast cancer recurrence and lung metastasis.

Medication Rules and Pharmaceutical Care of 875 Cases of COVID-19Treated by Traditional Chinese Medicine / 中国实验方剂学杂志

Objective:This paper analyzed the prescriptions of traditional Chinese medicine（TCM） for all patients diagnosed with novel coronavirus pneumonia（corona virus disease-2019，COVID-19）in Wuhan third hospital，analyzed the medication rules of the prescription，summarized the characteristics and thoughts of medication，and discussed the contents of TCM pharmaceutical care. Method:Use the data analysis software Excel 2007 and SQL server 2017 to perform statistical analysis for all inpatients in Wuhan Third Hospital from January 25，2020 to March 18，2020 who were treated by the prescriptions of TCM. The usage quantity，frequency，average dosage and dosage range of TCM were counted and compared with the dosage stipulated in the 2015 edition of Chinese Pharmacopoeia. Result:In this study，a total of 875 patients were included in the treatment prescription，involving 233 TCMs，and 20 high-frequency herbs were obtained，which were mainly used to dissolve phlegm，relieve cough and asthma，and tonify body. In the analysis of the characteristics of TCM，it is mainly composed of plain drugs，followed by cold drugs and warm drugs. The main taste of medicine is bitter，followed by spicy and sweet. The main meridians were the lung meridians，followed by the spleen and stomach meridians. By using association rule analysis and complex network analysis，it was found that the correlation degree of Artemisiae Scopariae Herba，Amomi Fructus Rotundus，Akebiae Caulis，Talcum and Fritillariae Cirrhosae Bulbus was very high，which could treat symptoms such as fever，cough，sputum，thirst，chest tightness and abdominal distension after COVID-19 infection. Conclusion:In this study，it was found that the core prescription for the treatment of COVID-19 in Wuhan Third Hospital was the addition and reduction of clearing damp agent，modified Ganlu Xiaodudan，and the reduction of expectorant modified Qingjin Jianghuotang，all the drugs were excess used than pharmacopoeia prescribed dosage. As a clinical Chinese pharmacist，we should distinguish the syndrome types according to the symptoms of the patients，and medication monitoring should be conducted from the aspects of usage and dosage of specific medication，processing product selection，compatibility，patient education，etc.