[Shenqi Wenfei Formula alleviates chronic obstructive pulmonary disease in rats with pulmonary qi deficiency syndrome by regulating NLRP3/GSDMD pyroptosis pathway].

OBJECTIVE: To investigate the effects of Shenqi Wenfei (SQWF) Formula on the NLRP3/GSDMD signaling pathway in pulmonary qi deficiency syndrome rats with chronic obstructive pulmonary diseases (COPD). METHODS: Rat models of COPD and lung qi deficiency syndrome induced by exposure to cigarette smoke and lipopolysaccharide (LPS) injection were randomized (n=8) for treatment with low-, medium- and high-dose SQWF or Yu Ping Feng (YPF). The changes in body weight, grip strength, lung function, pulmonary pathology, peripheral blood inflammatory cell counts, and levels of inflammatory factors in the bronchoalveolar lavage fluid (BALF) were examined. The expressions of proteins associated with the NLRP3/GSDMD signaling pathway in the lung tissues were determined with RT-qPCR and immunohistochemistry. RESULTS: The rat models of COPD and lung qi deficiency syndrome showed significantly reduced body weight, grip strength and lung function (P<0.01) with severe lung pathologies, increased levels of WBC, NEU% and MON% (P<0.01), decreased LYM% (P<0.01), and increased levels of TNF-α, IL-6, IL-1ß and IL-18 in the BALF (P<0.01); the mRNA and protein expression levels of NLRP3, ASC, GSDMD and IL-1ß all increased significantly in the lung tissue (P<0.01). Treatment with SQWF and YPF obviously increased body weight and improved grip strength, pulmonary function and lung pathology of the COPD rats (P<0.05). The treatments obviously improved the changes in peripheral blood inflammatory cell counts and inflammatory factors in the BALF of the rat models (P<0.01) and lowered the expressions of NLRP3, ASC, GSDMD and IL-1ß in the lung tissues (P<0.05) without significantly affecting the mRNA levels of caspase-1 and IL-18 (P>0.05), and the effects of SQWF were dose-dependent. CONCLUSION: SQWF Formula relieves inflammatory response and injury in the lung tissue of COPD rats with pulmonary qi deficiency syndrome possibly by inhibiting pyroptosis through regulating the NLRP3/GSDMD pathway.

Modular characteristics and mechanism of action of herbs for vascular calcification treatment in Chinese medicine: a data mining and network pharmacology-based identification.

OBJECTIVE: The aim of this study was to investigate the modular characteristics and mechanism of action of Chinese herbs for vascular calcification (VC) treatment. MATERIALS AND METHODS: Network pharmacology coupled with literature data mining was utilized to assess the Chinese herbal clinical performance as well as its similarity, characteristics, ingredient, target, and Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and network construction. RESULTS: The top 15 medications from the literature, according to the usage, and 190 active chemicals, 183 common targets between medication and VC-related targets were weeded out. Analysis of the relationships between the active ingredients, pharmacological targets, and signaling pathways helped to clearly define the therapeutic effect of Traditional Chinese Medicine (TCM). Importantly, we discovered seven most hub proteins (AKT1, CTNNB1, TNF, EGFR, TP53, JUN and IL-6) and two of the herbs' most fundamental ingredients (Formononetin and Luteolin) in TCM-mediated VC suppression. Mechanistically, the metabolic pathways [AGE-RAGE pathway, interleukin-17 (IL-17) pathway, and p53 pathway] as well as smooth muscle adaptation (functional remodeling) and oxidoreductase activity (redox homeostasis modulating) are also crucially implicated. CONCLUSIONS: Our work, accomplished by network pharmacology and data mining, increases our understanding of TCM in VC therapy and may offer insightful information for future drug discovery investigations.

[Association of nutritional status with clinical outcomes of stroke patients with acute anterior circulation large vessel occlusion after emergency endovascular treatment].

OBJECTIVE: To explore the influence of nutritional status on 90-day functional outcomes of stroke patients with acute large vessel occlusion in the anterior circulation after endovascular treatment (EVT). METHODS: We retrospectively analyzed the baseline, laboratory, surgical and 90-day follow-up data of patients with stroke resulting from acute large vessel occlusion in the anterior circulation, who underwent emergency endovascular treatment in our hospital from July, 2015 to December, 2020. A favorable outcome was defined as a modified Rankin scale score ≤2 at 90 days. Univariate and multivariate regression analyses were performed to explore the relationship between nutritional status and 90-day functional outcomes of the patients. RESULTS: A total of 459 patients (mean age of 68.29±11.21 years, including 260 males) were enrolled in this study. According to their prognostic nutritional index (PNI), the patients were divided into normal nutrition group (392 cases, 85.4%), moderate malnutrition group (44 cases, 9.6%), and severe malnutrition group (23 cases, 5.0%). Univariate analysis showed that the patients with good clinical outcomes had a lower proportion of malnutrition with a younger age, a lower rate of diabetes, lower baseline blood pressure, lower baseline NIHSS score, higher baseline ASPECT score, and higher rates of good collateral circulation and complete vascular recanalization. Multivariate analysis showed that in addition to age, diabetes, baseline systolic blood pressure, successful recanalization, baseline ASPECT score, baseline NIHSS score and collateral circulation, a greater PNI was a protective factor for a good 90-day outcome of patients after EVT (moderate vs severe: OR=0.245, 95% CI: 0.066-0.908, P=0.035; normal vs severe: OR=0.185, 95% CI: 0.059-0.581, P=0.004). CONCLUSION: Nutritional status an important factor affecting the 90-day outcomes after EVT of stroke patients with acute large vessel occlusion in the anterior circulation.

[Chaihu Guizhi Decoction plus or minus formula combined with capecitabine inhibits IL-6/STAT3 signaling to suppress triple-negative breast cancer xenografts in nude mice].

OBJECTIVE: To investigate the effect of Chaihu Guizhi Decoction (CHGZD) combined with capecitabine on growth and apoptosis of subcutaneous triple-negative breast cancer xenografts in nude mice and explore the possible mechanism. METHODS: Nude mouse models bearing subcutaneous triple-negative breast cancer xenografts were randomized into 6 groups (n=10) for treatment with distilled water (model group), low (10.62 g/kg), medium (21.23 g/kg) and high (42.46 g/kg) doses of CHGZD, capecitabine (0.2 mg/kg), or the combination of CHGZD (42.46 g/kg) and capecitabine (0.2 mg/k) once daily for 21 consecutive days. The general condition of mice was observed, and after 21-day treatments, the tumors were dissected for measurement of tumor volume and weight and histopathological examination with HE staining. Serum IL-6 levels of the mice were determined with enzyme-linked immunosorbent assay (ELISA), and the expression levels of IL-6, STAT3, p-STAT3, Bax, Bcl-2 and cyclin D1 in the tumor tissues were detected using real-time PCR and Western blotting. RESULTS: Compared with those in the model group, the tumor-bearing mice receiving treatments with CHGZD showed significantly increased food intake with good general condition, sensitive responses, increased body weight, and lower tumor mass (P < 0.01). Compared with capecitabine treatment alone, treatment with CHGZD alone at the medium and high doses and the combined treatment all resulted in significantly higher tumor inhibition rates (P < 0.01), induced obvious tumor tissue degeneration and reduced the tumor cell density. Treatments with CHGZD, both alone and in combination with capecitabine, significantly decreased serum IL-6 level, lowered the mRNA expression levels of IL-6 and STAT3, the protein expressions of IL-6, STAT3 and P-STAT3 (P < 0.05), and the mRNA and protein expressions of Bcl-2 and cyclin D1 (P < 0.05), and increased the mRNA and protein expressions of Bax in the tumor tissues (P < 0.05). CONCLUSION: CHGZD combined with capecitabine can significantly inhibit tumor growth in nude mice bearing triple-negative breast cancer xenografts, the mechanism of which may involve the inhibition of IL-6/STAT3 signaling pathway and regulation of Bax, Bcl-2 and cyclin D1 expressions to suppress tumor cell proliferation and differentiation and induce cell apoptosis.

[Qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets, a compound traditional Chinese herbal preparation].

OBJECTIVE: To conduct qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets, a compound preparation of traditional Chinese herbal medicine. METHODS: Thin-layer chromatography (TLC) was used for qualitative analysis of Tripterygium hypoglaucum in Yining Tablets and the analytical protocols were optimized. High-performance liquid chromatography (HPLC) was used to quantitatively analyze the content of triptolide (the main active ingredient of Tripterygium hypoglaucum) in Yinning Tablets. RESULTS: The results of TLC analysis showed that the test sample of Yinning Tablets and the positive control samples both produced clear, well separated spots without obvious interference in the blank samples. Assessment of the influences of the thin-layer plates from different manufacturers, temperature and humidity on the test results demonstrated good durability of the test. HPLC analysis of triptolide showed a good linear relationship within the concentration range of 1-100 µg/mL (regression equation: A=22.219C-19.165, r=0.9999); the contents of triptolide in 3 batches of Yinning tablets were 0.34, 0.34, and 0.28 µg per tablet, all within the range of 0.28-0.34 µg per tablet. It was finally determined that each Yinning tablet should not contain more than 0.6 µg of triptolide. CONCLUSION: TLC and HPLC are simple, accurate, durable and specific for qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets.

[Liuwei Buqi capsule modulates immune function by targeting multiple immune cell subsets in lung tissue of patients with COPD].

OBJECTIVE: To investigate the characteristics of immune cell subsets in the lung tissues of patients with chronic obstructive pulmonary disease (COPD) and the mechanism of Liuwei Buqi capsule in modulating immune and inflammatory imbalance in COPD. METHODS: We downloaded COPD-related single-cell RNA sequencing data from Gene Expression Omnibus (GEO) and identified COPD immune cell subsets using the Seurat package in the R software to construct an immune cell subsets-differential genes network. The target genes and active ingredients of Liuwei Buqi capsule were obtained from the Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), and the Liuwei Buqi capsule-immune cell subsets-target genes network was constructed by mapping the target genes to the differentially expressed genes in each immune cell subset. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to analyze significantly enriched pathways of the target genes, and the key genes involved in the top 20 pathways were identified. In a rat model of COPD, we investigated the effects of Liuwei Buqi capsule on pulmonary function, lung tissue pathology, serum levels of IL-1ß, NF-κB, and TNF-α, and expressions of IKBα, JNK, c-JUN, and c-FOS proteins in the lung tissue. RESULTS: A total of 18 immune-related cell subsets, including macrophages and alveolar macrophages, were identified in both COPD patients and healthy control subjects, and the patients with COPD showed significant changes the percentages of macrophages, cDC1, pDC, mast cells, T cells, and mature dendritic cells (P < 0.05). Liuwei Buqi capsules targeted multiple immune cell subsets, and the identified target genes were enriched mostly in such immune and inflammation-related signaling pathways as lipids and atherosclerosis, IL-17 signaling pathway, Toll-like receptor signaling pathway, and TNF signaling pathway; the genes CXCL8, IL1B, JUN, NFKBIA, MAPK8, and FOS were the key genes involved in the significantly enriched pathways. In the rat models of COPD, treatment with Liuwei Buqi capsule significantly improved pulmonary function, alleviated lung pathologies, reduced serum levels of IL-1ß, TNF-α, and NF-κB (P < 0.05) and pulmonary expressions of JNK, c-JUN, and c-FOS (P < 0.01) protein, and increased pulmonary expression of IκBα (P < 0.01). CONCLUSION: Liuwei Buqi capsule may play an immunomodulatory role by targeting multiple immune cell subsets in the lung tissue of COPD patients.

[Study on the relationship between ABCB1 gene polymorphism and hemorrhage risk after thrombolysis of cerebral ischemic stroke in Shangqiu area].

Using a cross-sectional study, 246 patients with hemorrhage and transformation after cerebral ischemic stroke(CIS) thrombolysis who were admitted to Shangqiu First People's Hospital, Shangqiu Municipal Hospital, and Shangqiu Liangyuan Traditional Chinese Medicine Hospital from March 2018 to May 2020 were selected as the observation group, 246 patients with no hemorrhage transformation after CIS thrombolysis during the same period were selected as the control group with a ratio of 1∶1. Polymerase chain reaction and pyrosequencing methods were used to detect the single nucleotide polymorphisms of the two groups of ABCB1 genes. The frequency distribution of each genotype of the two groups of ABCB1 gene polymorphism sites was counted. The conditional logistic regression equation was used to analyze the CIS after thrombolysis. Related influencing factors of hemorrhage transformation, and compare the single nucleotide polymorphisms of ABCB1 gene in patients with different prognosis in the observation group. The results showed that the CC genotype frequency of rs1045642 in the observation group was 34.55% higher than that of the control group 25.02%, the CT genotype frequency was 12.20%, and the TT genotype frequency 3.25% was lower than that of the control group 14.63% and 9.35% (χ2=21.527, P<0.05); GG genotype frequency at rs2032582 locus in observation group was 17.89%, GT genotype frequency 21.54% was lower than control group 37.60%, 93.96%, TT genotype frequency 10.57% higher than control group 2.44%, the difference was statistically significant (χ2=80.427, P<0.05); TT genotype at rs1045642 is a protective factor for hemorrhage transformation, and TT genotype at rs2032582 is a risk factor for hemorrhage transformation (OR=2.903, P<0.05). The risk of bleeding after thrombolysis in CIS patients in Shangqiu area may be related to the TT genotype at the ABCB1 rs1045642 locus and the TT genotype at the rs2032582 locus.

X-ray attenuation models to account for beam hardening in computed tomography.

We introduce a beam-hardening correction method for lab-based X-ray computed tomography (CT) by modifying existing iterative tomographic reconstruction algorithms. Our method simplifies the standard Alvarez-Macovski X-ray attenuation model [Phys. Med. Biol.21, 733 (1976)] and is compatible with conventional (i.e., single-spectrum) CT scans. The sole modification involves a polychromatic projection operation, which is equivalent to applying a weighting that more closely matches the attenuation of polychromatic X-rays. Practicality is a priority, so we only require information about the X-ray spectrum and some constants relating to material properties. No other changes to the experimental setup or the iterative algorithms are necessary. Using reconstructions of simulations and several large experimental datasets, we show that this method is able to remove or reduce cupping, streaking, and other artefacts from X-ray beam hardening and improve the self-consistency of projected attenuation in CT. When the assumptions made in the simplifications are valid, the reconstructed tomogram can even be quantitative.

[The efficacy and safety of different antimicrobial regimens in carbapenem-resistant Klebsiella pneumoniae bloodstream infections].

Objective: To evaluate the efficacy and safety of different antimicrobial regimens in patients with bloodstream infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP). Methods: The clinical date of patients with CRKP bloodstream infections were retrospectively analyzed at the First Affiliated Hospital of Zhejiang University Medical College between January 2017 and January 2018. All subjects were separated into three groups based on antibiotics regimens over 72 hours, including meropenem 2.0 g every 8 hours, tigecycline 200 mg as initial dose and 100 mg every 12 hours, and polymyxin B 1.25 mg/kg every 12 hours as salvage treatment of tigecycline. Results: A total of 86 patients were finally recruited, including 14, 52 and 20 patients in groups of meropenem, tigecycline and polymyxin B salvage, respectively. All of the strains were resistant to meropenem and susceptible to tigecycline and polymyxin B initially, while 2 of them became resistant to tigecycline during treatment. The 28-day mortality was significantly higher in meropenem group (13/14) than that in tigecycline group and polymyxin B salvage group (61.5%, 32/52) and (12/20), respectively (P<0.01), while as no significant difference was seen in the last two groups (χ(2)=0.014, P>0.05). The incidences of hepatic impairment [3.8%(2/52) vs. 1/20] and renal dysfunction (0 vs. 1/20) between tigecycline group and polymyxin B salvage group were both comparable (P>0.05). Conclusion: The meropenem-based therapy is not recommended for CRKP-related bloodstream infections. Tigecycline-based therapy is still disappointing despite salvage use of polymyxin B after 72 hours. Hepatic and nephretic toxicities caused by additional polymyxin B are acceptable.

Melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, hormonal profile and total antioxidant capacity: A placebo-controlled, double-blind trial.

In this study, we aimed at investigating the impact of melatonin supplementation on semen parameters, hormonal profile and total antioxidant capacity after varicocelectomy. Infertile male patients who were diagnosed with varicocele and underwent subinguinal varicocelectomy were included in the study. After performing subinguinal varicocelectomy, the patients were randomised into two groups: 27 receiving melatonin for 3 months and 27 as the placebo-controlled group receiving placebo for 3 months. The pre-operative parameters of semen analyses, hormonal profile and seminal oxidative stress status of both groups were compared with those of post-operative parameters. There were statistically significant improvements in post-operative parameters of semen analyses (sperm concentration, motility and proportions of normally formed spermatozoa), peripheral blood inhibin B and total antioxidant capacity in melatonin group compared with placebo group. In conclusion, melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, peripheral blood inhibin B and total antioxidant capacity; however, further studies including large number of samples are needed to make a proper decision on melatonin supplementation after varicocelectomy.

Immunoprotective effects of a hemin-binding peptide derived from hemagglutinin-2 against infection with Porphyromonas gingivalis.

The principal etiologic agent in periodontal disease, Porphyromonas gingivalis, generates cysteine proteases that bind heme with domains such as hemagglutinin-2 (HA2). High-affinity HA2-hemin binding supplies the porphyrin and ferric iron needed for growth and virulence. The DHYAVMISK peptide, recently identified at the hemin-binding site of HA2, inhibits hemin binding. We now evaluate the protective effect of vaccination with DGFPGDHYAVMISK (termed DK) against P. gingivalis using a rat infection model. Rats immunized with DK generated anti-peptide serum IgGs and salivary sIgAs (as measured by ELISA). In a subcutaneous abscess model, the protective effect of immunization was then investigated by measuring abscess size following subcutaneous injection with P. gingivalis. In an oral infection model, a ligature inoculated with P. gingivalis was used to induce periodontitis. The degree of bone erosion, ordinarily provoked by infection, was then evaluated by micro-computed tomography. We found that anti-peptide antibody titers of serum IgGs and salivary sIgAs for rats immunized with DK and adjuvant were significantly higher than for sham-immunized rats (injected with adjuvant/PBS alone; P < .05). In the subcutaneous abscess model, the DK + adjuvant-vaccinated rats recovered faster than sham-vaccinated animals, with their abscess sizes significantly smaller (P < .05). Further, in the experimental periodontitis model, bone loss at the molar palatal side for DK + adjuvant-vaccinated rats was significantly lower than for sham-vaccinated animals (P < .05). Collectively, these data demonstrate the potential of (DK) peptide immunization in terms of eliciting an immunoprotective effect against infection with P. gingivalis.

Cross-resistance between voriconazole and fluconazole for non-albicans Candida infection: a case-case-control study.

Cross-resistance (CR) between voriconazole and fluconazole for non-albicans Candida (NAC) species is not uncommon, but little is known about the risk factors and clinical consequences associated with this resistance phenotype. A case-case-control study was performed at a university-affiliated hospital in China between November 2012 and April 2016. The two case groups respectively comprised patients with a mono-resistance (MR) NAC infection (fluconazole or voriconazole resistance) and patients with a CR NAC infection (fluconazole and voriconazole resistance). Patients with a no-resistance (NR) NAC infection were included as the control group. Models were adjusted for demographic and clinical risk factors, and the risk of resistance associated with exposure to specific antibiotics or non-antibiotics were assessed. Of 259 episodes, 33 (12.7%) and 27 (10.4%) were identified as MR and CR NAC infections, respectively. The broad use of azoles was strongly associated with the emergence of MR and CR NAC infections (adjusted odds ratio [95% confidence interval] = 2.69 [1.10-6.58] and 2.53 [1.02-6.28], respectively). The time at risk (1.02 [1.00-1.03]) with 12 days as a breakpoint was also an independent risk factor for CR NAC infection. The number of species associated with a high minimum inhibitory concentration (≥128 µg/mL) of fluconazole was higher for CR NAC infections than for MR NAC infections. Different resistance phenotypes (CR vs. MR vs. NR) were associated with all-cause mortality rates. These findings indicate a worrisome propensity of CR NAC infections and emphasize the need for strict antifungal stewardship.

Serine hydroxymethyl transferase 1 stimulates pro-oncogenic cytokine expression through sialic acid to promote ovarian cancer tumor growth and progression.

High-grade serous (HGS) ovarian cancer accounts for 90% of all ovarian cancer-related deaths. However, factors that drive HGS ovarian cancer tumor growth have not been fully elucidated. In particular, comprehensive analysis of the metabolic requirements of ovarian cancer tumor growth has not been performed. By analyzing The Cancer Genome Atlas mRNA expression data for HGS ovarian cancer patient samples, we observed that six enzymes of the folic acid metabolic pathway were overexpressed in HGS ovarian cancer samples compared with normal ovary samples. Systematic knockdown of all six genes using short hairpin RNAs (shRNAs) and follow-up functional studies demonstrated that serine hydroxymethyl transferase 1 (SHMT1) was necessary for ovarian cancer tumor growth and cell migration in culture and tumor formation in mice. SHMT1 promoter analysis identified transcription factor Wilms tumor 1 (WT1) binding sites, and WT1 knockdown resulted in reduced SHMT1 transcription in ovarian cancer cells. Unbiased large-scale metabolomic analysis and transcriptome-wide mRNA expression profiling identified reduced levels of several metabolites of the amino sugar and nucleotide sugar metabolic pathways, including sialic acid N-acetylneuraminic acid (Neu5Ac), and downregulation of pro-oncogenic cytokines interleukin-6 and 8 (IL-6 and IL-8) as unexpected outcomes of SHMT1 loss. Overexpression of either IL-6 or IL-8 partially rescued SHMT1 loss-induced tumor growth inhibition and migration. Supplementation of culture medium with Neu5Ac stimulated expression of IL-6 and IL-8 and rescued the tumor growth and migratory phenotypes of ovarian cancer cells expressing SHMT1 shRNAs. In agreement with the ovarian tumor-promoting role of Neu5Ac, treatment with Neu5Ac-targeting glycomimetic P-3Fax-Neu5Ac blocked ovarian cancer growth and migration. Collectively, these results demonstrate that SHMT1 controls the expression of pro-oncogenic inflammatory cytokines by regulating sialic acid Neu5Ac to promote ovarian cancer tumor growth and migration. Thus, targeting of SHMT1 and Neu5Ac represents a precision therapy opportunity for effective HGS ovarian cancer treatment.

Study on the effects of blueberry treatment on histone acetylation modification of CCl4-induced liver disease in rats.

The objective of this study was to investigate the effects of blueberry treatment on histone acetylation modification of carbon tetrachloride (CCl4)-induced liver disease in rats. Laboratory rats were randomly divided into control, hepatic fibrosis, blueberry treatment, blueberry intervention, and natural recovery groups. Rats in the model groups were treated with CCl4 administered subcutaneously at 4- and 8-week intervals, and then executed. Both the 4- and 8-week treatment groups were treated with blueberry juice for 8 weeks, and then executed after 12 and 16 weeks, respectively. Following the experiment, four liver function and hepatic fibrosis indices were measured. Liver index was calculated, hematoxylin-eosin staining was conducted, and H3K9, H3K14, and H3K18 expressions were evaluated among the nuclear proteins of the liver tissues. No differences in alanine transaminase were noted between the control and intervention groups, but significant differences were detected among the model, treatment, and natural recovery groups (P < 0.01). Significant differences were also observed in aspartate transaminase, hyaluronic acid, and collagen IV among the model, treatment, intervention, and natural recovery groups (P < 0.01, P < 0.01, P < 0.01). Liver index, and H3K9 and H3K14 expression were significantly different among the model groups (P < 0.05 and P < 0.01), whereas H3K18 expression was dramatically different among model, treatment, intervention, and natural recovery groups (P < 0.01). Following blueberry treatment, rat liver function and hepatic fibrosis improved, potentially indicating that blueberry components could regulate histone acetylation and improve liver pathologic changes in rats with CCl4-induced disease.

[Preventive and therapeutic effects of compound ginkgo extract in rats with nonalcoholic steatohepatitis induced by high-fat, high-fructose diet].

Objective: To establish a rat model of nonalcoholic steatohepatitis (NASH), and to investigate the preventative and therapeutic effects of compound ginkgo extract against NASH. Methods: A total of 60 male Sprague-Dawley rats were fed with high-fat feed and 10% fructose water for 24 weeks to establish the rat model of NASH. The general behaviors of the rats were observed, and the body weight was recorded. Blood samples from the inferior vena cava and the liver were collected after the last administration to measure serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), as well as liver function parameters. The liver index was calculated, HE staining was performed to observe liver histopathological changes, and the total lipase activity and the levels of TC, TG, and free fatty acid (FFA) in liver tissue were measured. Results: After 24 weeks, compared with the normal group, the model group had a significantly faster increase in body weight, significant increases in serum levels of TC (2.20±0.52 mmol/L), TG (0.87±0.22 mmol/L), LDL-C (1.22±0.50 mmol/L), alanine aminotransferase (ALT) (129.4±44.7 U/L), and aspartate aminotransferase (AST) (209.3±42.8 U/L), liver index (3.62%±0.28%), and the levels of TC (4.42±1.39 mmol/mg.prot), TG (0.85±0.11 mmol/mg.prot), and FFA (644.78±36.65µmol/L) in liver tissue, and significant reductions in serum HDL-C level (0.58±0.11 mmol/L) and the activity of lipoprotein lipase (LPL) (9.95±1.64 U/mg.prot) and hepatic lipase (HL) (9.91±1.03 U/mg.prot) (allP< 0.01). In addition, the pathological results showed severe hepatocyte steatosis, varying degrees of inflammatory cell infiltration, exudation in the portal area, and necrosis of liver cells in the model group. After the intervention with compound ginkgo extract, there were significant reductions in serum levels of TC (1.78±0.21 mmol/L), TG (0.58±0.07 mmol/L), LDL-C (0.84±0.19 mmol/L), and ALT (84.1±17.1 U/L), AST (155.4±20.9 U/L), liver index (2.71%±0.15%), and the levels of TC (2.24±1.02 mmol/mg.prot), TG (0.46±0.11 mmol/mg.prot), and FFA (580.56±50.63µmol/L) in liver tissue, as well as significant increases in serum HDL-C level (0.68±0.10 mmol/L) and the activities of LPL (15.54±2.21 U/mg.prot) and HL (11.92±1.87 U/mg.prot) (P< 0.05 orP< 0.01). At the same time, it significantly reduced hepatomegaly in rats and improved fatty degeneration and degree of inflammation in liver cells. Conclusion: Compound ginkgo extract can prevent and treat NASH by correcting dyslipidemia, improving liver function and fatty degeneration in hepatocytes, and reducing the degree of inflammation, and its mechanism of action may be associated with increasing total lipase activity, reducing FFA in the liver, increasing the decomposition of TG, and reducing the synthesis of TG.

[Refractory allergic rhinitis serum levels of vitamin D and vitamin D calcium chewable therapeutic value].

Objective:To survey clinical efficacy of vitamin D calcium chewable tablets in the treatment of adults with refractory allergic rhinitis and investigate the level of 1,25-dihydroxyvitamin D3 in the patients' serum.Method:Sixty cases who were dignosed with adult refractory moderate to severe allergic in our hospital outpatient were admitted in our study from May 2014 to November 2015.Sixty cases of healthy people in our hospital medical center were selected as the control group,serum 1,25-dihydroxyvitamin D3 were detected in all patients and normal controls before the treatment.The control group were treated using fluticasone propionate nasal spray and left cetirizine while the experimental group were added with vitamin D calcium chewable tablets on the basis of the control group. Before treatment and after 2 or 4 weeks of treatment,a visual analog scale of the overall classification of nasal symptoms were used of and clinical symptoms evaluation.Result:Compared with healthy controls,the incidence of serum 1,25-dihydroxy vitamin D3 content is:(17.46±4.94)µg/L and the difference was statistically significant(P<0.0).The symptom scores of the treatment group and control group were significantly decreased before treatment and the difference had statistical significance(P<0.05).After four weeks of treatment,the experimental group of the symptom scores were lower than that of control group.The differences were statistically significant(P<0.05).Conclusion:Serum 1,25 dihydroxyvitamin D3 deficiency is a widespread phenonmenon in refractory allergic rhinitis.Vitamin D calcium chewable tables adjuvant treatment of allergic rhinitis is one of methods with safe,effective,and simple treatment.

CpG oligonucleotides and Astragalus polysaccharides are effective adjuvants in cultures of avian bone-marrow-derived dendritic cells.

1. The potential use of CpG oligodeoxynucleotides and/or Astragalus polysaccharide (APS) as adjuvants for the culture of chicken bone-marrow-derived dendritic cells (chBM-DCs) was investigated. 2. Chicken dendritic cells (DCs) were isolated and cultured in the presence of recombinant chicken granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. The chBM-DC displayed typical DC morphology and expressed DC surface markers (MHC-II and CD11c). 3. Cultured chBM-DC showed effective T-cell activation in vitro, based on a mixed lymphocyte response (MLR). Flow cytometry analysis showed an increased proportion of cells expressing CD40 and CD80 in the APS-stimulated culture, compared to the control culture. In the MLR, the APS- and CpG-stimulated chBM-DC could activate T-cells more than control chBM-DC. Real-time PCR assays showed that CpG can activate the TLR21 and an inflammatory response, while APS just reduced the expression of IRF-3. 4. The results demonstrated that in vitro the adjuvant CpG can stimulate chBM-DC to mature by activation of the TLR-signalling pathway, whereas the adjuvant APS stimulates maturation of chBM-DC in vitro to a lesser degree and by another mechanism.

Isolation and characterization of a chalcone isomerase gene promoter from potato cultivars.

Chalcone isomerase (CHI) is a key enzyme involved in anthocyanin metabolism. Previous research on CHI has mainly focused on cDNA cloning and gene expression. In the current study, the 1425-bp potato CHI promoter (PCP) was isolated from four potato cultivars (Heijingang, Zhongshu 7, Désirée, and Favorita) using PCR and DNA sequencing. The PCP contained many cis-regulatory elements (CREs) related to anthocyanin metabolism, tissue specificity, light response, stress, and hormone induction. Of the PCP CREs identified, 19 were common to those found in the higher plants examined, based on plant CRE databases. Multiple sequence alignment showed six single nucleotide variation sites in PCP among the potato cultivars examined, resulting in changes in the number of CREs connected with tissue specificity, anthocyanin metabolism, and light response. The 665-bp PCP fragments from Favorita and 1425-bp PCP fragments from Heijingang were used to construct plant expression vectors, which may be a useful tool for biological engineering. A transient expression assay demonstrated that the two PCP fragments from Heijingang could direct the expression of a green fluorescent protein gene in onion epidermis and a ß-glucuronidase gene in all potato tuber tissues with different colors, suggesting that the single nucleotide variation in the PCP did not affect its activity, and that silencing of the CHI gene in Favorita may be attributed to other regulatory factors.

Structure and function of soil microbial community in artificially planted Sonneratia apetala and S. caseolaris forests at different stand ages in Shenzhen Bay, China.

The present study examined the relationships between soil characteristics, microbial community structure and function in the forests artificially planted with exotic Sonneratia apetala at stand ages of 1-, 2-, 7-, 10- and 14-years and Sonneratia caseolaris of 1-, 4-, 7-, 10- and 14-years in Futian National Nature Reserve, Shenzhen Bay, China. The 7-years old forests of both Sonneratia species reached peak growth and had the highest content of nitrogen and phosphorus, enzymatic activities, including dehydrogenase, cellulase, phosphatase, urease and ß-glucosidase, except arylsulphatase which increased continuously with stand ages. The microbial community structure reflected by phospholipid fatty acid (PLFA) profiles also reached the maximum value in the 7-years old forests and soil bacterial PLFAs in both forests were significantly higher than fungal PLFAs. The canonical correlation analysis revealed that differences in microbial structural variables were significantly correlated to the differences in their functional variables, and the highest correlation was found between the soil enzymatic activities and the content of carbon and nitrogen.

Inheritance and fine mapping of a restorer-of-fertility (Rf) gene for the cytoplasmic male sterility in soybean.

The cytoplasmic male sterility (CMS) line FuCMS5A and its restorer line FuHui9 were crossed to produce a segregating F(2) population for pollen fertility assay and the genetic mapping of restorer-of-fertility (Rf) gene. Results showed that the individual F(2) plants were fertile or semi-fertile based on their pollen fertility characteristics. The average ratios of viable pollen were 96.90% and 50.00% for each class of individuals. The segregation of F(2) plants showed a good fit to a 1:1 ratio, which reflects a typical heredity pattern of gametophytic CMS with fertility restorer being controlled by a single dominant gene. Using bulk segregation analysis (BSA) and genetic mapping, the Rf gene was mapped on molecular linkage group J (chromosome 16), between the simple sequence repeat (SSR) makers BARCSOYSSR-16-1064 and BARCSOYSSR-16-1082 with the distances of 0.59 and 0.83 cM, respectively. Four SSR markers (BARCSOYSSR-16-1070, Sctt011, BARCSOYSSR-16-1076 and BARCSOYSSR-16-1077) were cosegregating with this Rf gene in the mapping population. These makers will greatly facilitate the maker assisted selection procedures in CMS breeding programs and it lays a foundation for further map-base cloning of the Rf gene.