Effect of acupuncture on regulating IL-17, TNF-ɑ and AQPs in Sjögren's syndrome.

AIM: The aim of the study was to observe the effect of acupuncture on regulating interleukin (IL)-17, tumor necrosis factor (TNF)-ɑ, and aquaporins (AQPs) in Sjögren's syndrome (SS) on patients and on non-obese diabetic (NOD) models. METHODS: Levels of anti-AQP 1, 5, 8, and 9 antibodies, IL-17, and TNF-ɑ in the serum of SS patients were compared prior and following 20 acupuncture treatment visits during 8 weeks. While in murine model, five groups were divided to receive interventions for 4 weeks, including control, model, acupuncture, isoflurane, and hydroxychloroquine. The submaxillofacial gland index, histology, immunohistochemistry of AQP1, 5, salivary flow, together with IL-17, and TNF-ɑ expression in peripheral blood were compared among the groups. RESULTS: Acupuncture reduced IL-17, TNF-ɑ, and immunoglobin A levels, and numeric analog scale of dryness in 14 patients with SS (p < 0.05). The salivary flow was increased, and the water intake decreased in NOD mice receiving acupuncture treatments. IL-17 and TNF-ɑ levels in peripheral serum were down-regulated (p < 0.05) and AQP1, 5 expression in the submandibular glands up-regulated in mice. CONCLUSION: The effect on relieving xerostomia with acupuncture may be achieved by up-regulating the expression of AQP1. AQP5, down-regulating levels of IL-17 and TNF-ɑ, and a decrease in inflammation of glands.

Lupus Recipe inhibits cGVHD-induced lupus nephritis in mice and promote renal LC3-associated autophagy.

Lupus nephritis (LN) is one of the most severe manifestations of systemic lupus erythematosus (SLE). The chronic graft versus host disease (cGVHD) mouse model is a well-established model of SLE. LC3-associated autophagy plays a critical role in extracellular particle clearance, including pathogens and apoptotic cells. Lupus Recipe (LR) is a Chinese herbal compound that has been proven to be effective in treating SLE. In the study, we investigated the protective effects of LR or LR combined with prednisone on cGVHD mouse model and LC3-associated autophagy in the kidney. The mice were subjected to six groups. The LR treatment group received LR at the dosage of 1.15 and 2.3 g/kg/day, respectively. The corticosteroid treatment group received prednisone at a dosage of 5 mg/kg/day. The combination treatment group received LR at a dosage of 2.3 g/kg/day, and prednisone at 2.5 mg/kg/day. LR treatment reduced proteinuria and serum triglyceride levels, as well as spleen weight. LR also alleviated pathologic damage and immunoglobulin G deposition in the kidney. LR combined with a low dose of prednisone significantly improved kidney function and decreased serum triglyceride, total cholesterol, and spleen weight. In addition, combination treatment relieved kidney injury more effectively than LR alone. Western blot revealed that LR treatment or LR combined with prednisone increased the LC3-associated autophagy protein of Rubicon and Nox2, as well as LC3I levels in the kidney tissues. In conclusion, LR inhibited the manifestation of cGVHD-induced LN, which may attribute to the increased levels of LC3-associated autophagy.

Neonatal-onset multiple acyl-CoA dehydrogenase deficiency (MADD) in the ETFDH gene: A case report and a literature review.

RATIONALE: Multiple acyl-CoA dehydrogenase deficiency (MADD) is a rare inborn error of metabolism affecting fatty acid, amino acid, and choline metabolism. The clinical manifestation of MADD is heterogeneous, from severe neonatal forms to mild late-onset forms. PATIENT CONCERNS: Here, we report a patient who presented with severe hypoglycemia and exercise intolerance suggestive of MADD. Serum tandem mass spectrometry analysis indicated elevated levels of various acyl carnitines at 25 days of age. Exome sequencing of the proband revealed compound heterozygous mutations, c. 413T>G (p.Leu138Arg) and c.1667C > G (p.Pro556Arg), in the ETFDH gene as the probable causative mutations. DIAGNOSES: Based on the patient's clinical presentation and test results, the patient was diagnosed with MADD. INTERVENTIONS: A high-calorie and reduced-fat diet was given together with oral supplements of L-carnitine (150 mg/day). OUTCOMES: He passed away at the age of 4 months because of severe respiratory distress accompanied by muscle weakness. LESSONS: He passed away at the age of 4 months because of severe respiratory distress accompanied by muscle weakness. Clinicians should consider MADD in the differential diagnosis when patients present with muscle weakness and biochemical abnormalities. Gene testing plays a critical role in confirming the diagnosis of MADD and may not only prevent the need for invasive testing but also allow for timely initiation of treatment.

Molluscicidal effectiveness of Luo-Wei, a novel plant-derived molluscicide, against Oncomelania hupensis, Biomphalaria alexandrina and Bulinus truncatus.

BACKGROUND: Control of snail intermediate hosts has been proved to be a fast and efficient approach for interrupting the transmission of schistosomiasis. Some plant extracts have shown obvious molluscicidal activity, and a new compound Luo-Wei, also named tea-seed distilled saponin (TDS), was developed based on the saponins extracted from Camellia oleifera seeds. We aimed to test the molluscicidal activity of 4% TDS against the intermediate host snails in China and Egypt, and evaluate its environmental safety to non-target organisms. METHODS: In the laboratory, Oncomelania hupensis, Biomphalaria alexandrina and Bulinus truncatus were exposed to 4% TDS, and the median lethal concentration (LC50) was estimated at 24, 48 and 72 h. In the field, snail mortalities were assessed 1, 2, 3 and 7 d post-immersion with 2.5 g/m3 4% TDS and 1, 3, 7 and 15 d post-spraying with 5 g/m2 4% TDS. In addition, the acute toxicity of 4% TDS to Japanese quail (Coturnix japonica), zebrafish (Brachydanio rerio) and freshwater shrimp (Macrobrachium nipponense) was assessed by estimations of LC50 or median lethal dose (LD50). RESULTS: In the laboratory, the LC50 values of 4% TDS for O. hupensis were 0.701, 0.371 and 0.33 mg/L at 24, 48 and 72 h, respectively, and 4% TDS showed a 1.975 mg/L [corrected] 24 h LC50 against B. alexandrina, and a 1.396 mg/L 24 h LC50 against B. truncatus. Across all study regions, the pooled mortalities of O. hupensis were 72, 86, 94 and 98% at 1, 2, 3 and 7 d, following field immersion of 4% TDS at a dose of 2.5 g/m3, and were 69, 77, 85 and 88% at 1, 3, 7 and 15 d, following field spraying at 5 g/m2, respectively. 4% TDS had moderate toxicity to Japanese quail (7 d LD50 > 60 mg/kg) and to shrimp (96 h LC50 = 6.28 mg/L; 95% CI: 3.53-11.2 mg/L), whereas its toxicity to zebrafish was high (96 h LC50 = 0.15 mg/L; 95% CI: 0.14-0.17 mg/L). CONCLUSIONS: 4% TDS is active against O. hupensis, B. alexandrina and B. truncatus under laboratory and field conditions, and it may be a candidate molluscicide of plant origin.

Sonographic diagnosis of colorectal polyps in children: Diagnostic accuracy and multi-factor combination evaluation.

This study was established to evaluate the diagnostic value of ultrasonography in screening colorectal polyps in children and to discuss the necessity of colonic preparation before an ultrasonic examination.In this study, 288 children with colorectal polyps managed at our hospital between January 2007 and December 2016 were retrospectively reviewed. All patients were examined before and after basic colon preparation. The colorectal polyps were confirmed by colonoscopy/laparotomy and histopathology. Among all 288 patients, solitary polyps were identified in 278 patients (96.52%), and multiple polyps were identified in 10 patients (43 polyps) (3.48%) by colonoscopy/laparotomy and histopathology.By ultrasonic examination, 264 cases (264/278) were detected as solitary polyp and 9 cases (9/10) as multiple polyps (31 polyps). In 278 solitary polyps, 180 (64.74%) were detected by ultrasonic examination without a colon preparation. Following glycerine enema (10-20 mL) treatment, 264 (94.96%) cases were detected by ultrasonic examination. The sensitivity and specificity of ultrasonography with glycerine enema for the detection of colorectal polyps were 94.96% and 100%, respectively. Colon preparation significantly increased the proportion of polyps identified by ultrasonography (P < .0001), as well as the diagnostic rate of polyps in rectum, sigmoid colon and descending colon (P < .05).Ultrasonography can be the primary diagnostic method for screening colorectal polyps in children on the strength of its safety, validity, and accuracy. Basic colon preparation with glycerine enema is recommended for children, which enable the detection of intraluminal lesions before ultrasonic examination.

Enhanced lincomycin production by co-overexpression of metK1 and metK2 in Streptomyces lincolnensis.

Streptomyces lincolnensis is generally utilized for the production of lincomycin A (Lin-A), a clinically useful antibiotic to treat Gram-positive bacterial infections. Three methylation steps, catalyzed by three different S-adenosylmethionine (SAM)-dependent methyltransferases, are required in the biosynthesis of Lin-A, and thus highlight the significance of methyl group supply in lincomycin production. In this study, we demonstrate that externally supplemented SAM cannot be taken in by cells and therefore does not enhance Lin-A production. Furthermore, bioinformatics and in vitro enzymatic assays revealed there exist two SAM synthetase homologs, MetK1 (SLCG_1651) and MetK2 (SLCG_3830) in S. lincolnensis that could convert L-methionine into SAM in the presence of ATP. Even though we attempted to inactivate metK1 and metK2, only metK2 was deleted in S. lincolnensis LCGL, named as ΔmetK2. Following a reduction of the intracellular SAM concentration, ΔmetK2 mutant exhibited a significant decrease of Lin-A in comparison to its parental strain. Individual overexpression of metK1 or metK2 in S. lincolnensis LCGL either elevated the amount of intracellular SAM, concomitant with 15% and 22% increase in Lin-A production, respectively. qRT-PCR assays showed that overexpression of either metK1 or metK2 increased the transcription of lincomycin biosynthetic genes lmbA and lmbR, and regulatory gene lmbU, indicating SAM may also function as a transcriptional activator. When metK1 and metK2 were co-expressed, Lin-A production was increased by 27% in LCGL, while by 17% in a high-yield strain LA219X.

QGQS Granule in SHR Serum Metabonomics Study Based on Tools of UPLC-Q-TOF and Renin-Angiotensin-Aldosterone System Form Protein Profilin-1.

QGQS granule is effective for the therapeutic of hypertension in clinic. The aim of this research is to observe the antihypertension effect of QGQS granule on SHR and explain the mechanism of its lowering blood pressure. 30 SHR were selected as model group, captopril group, and QGQS group, 10 WKYr were used as control group, and RBP were measured on tail artery consciously. And all the serum sample analysis was carried out on UPLC-TOF-MS system to determine endogenous metabolites and to find the metabonomics pathways. Meanwhile, ELISA kits for the determination pharmacological indexes of PRA, AngI, AngII, and ALD were used for pathway confirmatory; WB for determination of profilin-1 protein expression was conducted for Ang II pathway analysis as well. It is demonstrated that QGQS granule has an excellent therapeutic effect on antihypertension, which exerts effect mainly on metabonomics pathway by regulating glycerophospholipid, sphingolipid, and arachidonic acid metabolism, and it could inhibit the overexpression of the profilin-1 protein. We can come to a conclusion that RAAS should be responsible mainly for the metabonomics pathway of QGQS granule on antihypertension, and it plays a very important role in protein of profilin-1 inhibition.

Identification of novel PAD4 inhibitors based on a pharmacophore model derived from transition state coordinates.

1.4 Protein arginine deiminases 4 (PAD4) is an attractive target for the development of novel and selective inhibitors of Rheumatoid Arthritis (RA). F-amidine is known as mechanism-based inhibitor targeting PAD4 and used as inactivators by covalently modifying the active site Cys645. To identify novel structural inhibitors of PAD4, we investigated the flexibility of protein on basis of the transition state geometry of PAD4 inhibited by F-amidine from our previous QM/MM calculation. And a pharmacophore model was generated containing four features (ADHH) using five representative structures from molecular dynamic (MD) simulation on basis of the transition state geometry of PAD4 inhibited by F-amidine. We performed virtual screening using the pharmacophore model and molecular docking methods, resulting in the discovery of two molecules with KD (dissociation equilibrium constant) values of 112µM and 218µΜ against PAD4 through Surface Plasmon Resonance (SPR) experiments. These two molecules could potentially serve as PAD4 inhibitors.

Effects of vegetative propagule pressure on the establishment of an introduced clonal plant, Hydrocotyle vulgaris.

Some introduced clonal plants spread mainly by vegetative (clonal) propagules due to the absence of sexual reproduction in the introduced range. Propagule pressure (i.e. total number of propagules) may affect the establishment and thus invasion success of introduced clonal plants, and such effects may also depend on habitat conditions. A greenhouse experiment with an introduced plant, Hydrocotyle vulgaris was conducted to investigate the role of propagule pressure on its invasion process. High (five ramets) or low (one ramet) propagule pressure was established either in bare soil or in an experimental plant community consisting of four grassland species. H. vulgaris produced more total biomass under high than under low propagule pressure in both habitat conditions. Interestingly, the size of the H. vulgaris individuals was smaller under high than under low propagule pressure in bare soil, whereas it did not differ between the two propagule pressure treatments in the grassland community. The results indicated that high propagule pressure can ensure the successful invasion in either the grass community or bare soil, and the shift in the intraspecific interaction of H. vulgaris from competition in the bare soil to facilitation in the grassland community may be a potential mechanism.

[Protective effect of combined administration blood-activating drug and sedative drug on acute myocardial infarction rats].

OBJECTIVE: To observe the protective effect of combined administration of blood-activating drug and sedative drug on myocardial injury of acute myocardial infarction (AMI) rats. METHOD: The acute myocardial infarction (AMI) model was established by occluding the left descending coronary artery of Wistar rats. These rats were further divided into four groups (n = 15 per group): the sham-operated group, the AMI model group, the blood-activating drug group and the combined administration group. RESULT: Compared with the sham-operated group, the AMI model group showed significant decrease in ejection fraction (EF) and fractional shortening (FS) (P < 0.001) and notable increase in the left ventricular end-diastolic internal diameter (LVIDd) and left ventricular end-systolic internal diameter (LVIDs) (P < 0.01), with the infarct area of left ventricular front wall up to about 70%-90%. Besides, tissue was severely replaced by collagen deposition and fibrosis, the sarcomeres disorganized and mitochondrial abnormalized. Compared with the AMI model group, the blood-activating drug group and the combined administration group showed significant increase in the values of EF and FS (P < 0.05 or P < 0.01) and obvious reduction in LVIDd and LVIDs (P < 0.05 or P < 0.01), with the infarction area of left ventricular front wall up to about 40%-60%. The collagen deposition and myocardium fibrosis, the disorganized sarcomeres and mitochondrial abnormalities relieved significantly. And compared with blood-activating drug group, the combined administration group demonstrated further increase in the values of EF and FS and further decrease in LVIDd and LVIDs (P < 0.05), the collagen deposition and myocardium fibrosis, the disorganized sarcomeres and mitochondrial abnormalities relieved even more in Huoxue plus Anshen prescription group. CONCLUSION: The combined administration of blood-activating drug and sedative drug can further improve cardiac structure and function after myocardial ischemia infarction and have an obvious synergistic effect which may be related to sedative drug's effect of resisting lipid peroxide, stabling myocardial cell membrane and mitochondrial membrane and relieving cardiac cell injury.

Facile synthesis of 1,2,3-triazole analogs of SGLT2 inhibitors by 'click chemistry'.

Novel analogs of SGLT2 inhibitors containing the 1,2,3-triazole motif were designed and synthesized for urinary glucose excretion evaluation. The C-glucosides with triazole aglycone can be easily constructed by click chemistry. Most of the synthesized compounds increased urinary glucose excretion and demonstrated inhibition of glucose transport.

Salvianolic acid B inhibits the TLR4-NFκB-TNFα pathway and attenuates neonatal rat cardiomyocyte injury induced by lipopolysaccharide.

OBJECTIVE: To investigate the role of the TLR4-NFκB-TNFα inflammation pathway on: lipopolysaccharide (LPS)-induced neonatal rat cardiomyocyte injury and the possible protective effects of salvianolic acid B (Sal B). METHODS: Wistar rat (1-2 days old) cardiomyocytes were isolated and cultured. Sal B 10(-5)mol/L, 10(-6)mol/L and 10(-7)mol/L were pre-treated for 6 h in the culture medium. LPS (1 µg/mL) was added to mol/the culture medium and kept for 6 h to induce inflammation injury. The concentration of lactate dehydrogenase (LDH) in the supernatant was detected by spectrophotometry. The concentrations of tumor necrosis factor α (TNFα) and heat shock protein 70 (HSP70) in the supernatant were detected by enzyme linked immunosorbent assay. The protein expressions of toll, such as receptor 4 (TLR4) and nuclear factor kappa B (NFκB) were detected by immunohistochemistry. The mRNA expressions of TLR4 and NFκB were detected by real-realtime reverse transcription polymerase chain reaction (RT-PCR). RESULTS: (1) The concentrations of LDH and: TNFα in the LPS control group were significantly higher than those in the control group (561.41±67.39 U/L and 77.94±15.08 pg/mL, versus 292.13±26.02 U/L and 25.39±16.53 pg/mL, respectively, P<0.01, P<0.05). Compared with the LPS control group, the concentrations of LDH and TNFα were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (451.76±83.96 U/L and 34.00±10.38 pg/mL, respectively, P<0.05). (2) The TLR4 and NFκB protein expression area in the LPS control group were significantly higher than those in the control group (1712.41±410.12 µm(2) and 2378.15±175.29 µm(2), versus 418.62±24.42 µm(2) and 1721.74±202.87 µm(2), respectively, P<0.01). The TLR4 and NFκB protein expression internal optical density (IOD) values in the LPS control group were also significantly higher than those in the control group (3.06±0.33 and 7.20±1.04, versus 0.91±0.21 and 4.24±0.48, respectively, P<0.05 and P<0.01). Compared with the LPS control group, the TLR4 and NFκB protein expression areas were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (1251.54±133.82 µm(2) and 1996.37±256.67 µm(2), respectively, P<0.05), the TLR4 and NFκB protein expression IOD values were also significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.92±0.28 and 5.17±0.77, respectively, treated P<0.05). (3) The TLR4 and NFκB mRNA expressions (2(-ΔΔ)CT value) in the LPS control group were significantly higher than those in the control group (3.16±0.38 and 5.03±0.43 versus 1.04±0.19 and 1.08±0.21, respectively, P<0.01). Compared with the LPS control group, the TLR4 and NFκB mRNA expressions (2(-ΔΔ) -CT value) were significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.34±0.22 and 1.74±0.26, respectively, treated P<0.05). The concentration of HSP70 did not show any 0.05). CONCLUSIONS: The TLR4-NFκB-TNFα pathway was quickly activated: and was independent of HSP70 in the early phase of neonatal cardiomyocyte injury induced by LPS. The protective effects of Sal B may be through inhibiting the TLR4-NFκB-TNFα pathway and are dose-dependent.

[Effects of Chinese herbal medicine Yiqi Huoxue Formula on TGF-ß/smad signal transduction pathway and connective tissue growth factor in rats with renal interstitial fibrosis].

OBJECTIVE: To observe the effects of Yiqi Huoxue Formula (YQHXF), a compound Chinese herbal medicine, on transforming growth factor-ß (TGF-ß)/smad signal transduction pathway and connective tissue growth factor (CTGF) in rats with renal interstitial fibrosis METHODS: Unilateral ureteral obstruction (UUO) rat model was established and the rats were randomly divided into 5 groups: untreated group, high-, medium-, and low-dose YQHXF groups and fosinopril sodium group. Another group with sham operation was set as control. All rats were administered with corresponding drugs for 3 weeks. After the last administration, each rat was sacrificed and weighed and the serum was separated for creatinine (Cr) and blood urea nitrogen (BUN) detection. Kidneys of the rats were taken out, and mRNA and protein expressions of TGF-ß, smad2, smad7 and CTGF were measured with real-time fluorescent quantitative reverse transcription-polymerase chain reaction and Western blotting respectively; fibrosis of the kidney tissue was observed with hematoxylin-eosin (HE) staining and Masson trichrome staining. RESULTS: Compared with sham-operation group, Cr and BUN in serum of UUO groups were increased, while high-dose YQHXF treatment decreased the UUO-induced increase of Cr and BUN levels. HE staining and Masson staining results showed that the renal tubular epithelial cells in untreated group got atrophied; lumens of renal tubules expanded; fibroplastic proliferation and inflammatory cell infiltration were observed in renal interstitium; the number of glomerulus decreased and collagen increased significantly compared with sham-operation group. In the high- and medium-dose YQHXF groups and fosinopril sodium group, the histopathological changes of inflammatory cell infiltration, fibroplastic proliferation, expansion of lumens of renal tubules was improved as compared with the untreated group. The mRNA and protein expressions of TGF-ß, smad2 and CTGF in untreated group were higher than those in sham-operation group (P<0.05), and the mRNA and protein expressions of smad7 in untreated group were lower than those in the sham-operation group (P<0.05). Compared with untreated group, high- and medium-dose of YQHXF significantly down-regulated the mRNA and protein expressions of TGF-ß, smad2 and CTGF (P<0.01, P<0.05), and up-regulated the mRNA and protein expressions of smad7 (P<0.01, P<0.05). CONCLUSIONS: The mRNA expression of CTGF in UUO rats may be regulated by TGF-ß/smad signaling transduction pathway. YQHXF might inhibit the expression of CTGF through down-regulation of TGF-ß and smad2 and up-regulation of smad7, thus inhibiting the progression of renal interstitial fibrosis.