[Genome-wide Mendelian randomization study of the pathogenic role of gut microbiota in benign biliary tract diseases].

Objective: To investigate the causal relationship between intestinal flora and benign biliary diseases by genome-wide Mendelian randomization. Methods: This is a retrospective observational study. The data from the genome-wide association study of the gut microbiota from 18 340 samples from the MiBioGen consortium were selected as the exposure group,and the data from the genome-wide association study of biliary tract diseases were obtained from the FinnGen consortium R8 as the outcome group. There were 1 491 cases of primary sclerosing cholangitis,32 894 cases of cholelithiasis,3 770 cases of acalculous cholecystitis,and 34 461 cases of cholecystitis. Single nucleotide polymorphisms were screened as instrumental variables,and the Mendelian randomization method was used to infer the causal relationship between exposures and outcomes. The inverse variance weighting method (IVW) was used as the main basis, supplemented by heterogeneity,pleiotropy and sensitivity tests. Results: Coprococcus 2 was associated with a reduced risk of cholelithiasis (IVW OR=0.88,95%CI:0.80 to 0.97,P=0.012) and cholecystitis (IVW OR=0.88,95%CI:0.80 to 0.97,P=0.011). Coprococcus 3 was associated with cholelithiasis (IVW OR=1.15,95%CI:1.02 to 1.30,P=0.019) and acalculous cholecystitis(IVW OR=1.48, 95%CI: 1.08 to 2.04,P=0.016) and cholecystitis (IVW OR=1.17, 95%CI: 1.02 to 1.33, P=0.020). Peptococcus was associated with an increased risk of cholelithiasis (IVW OR=1.08, 95%CI:1.02 to 1.13, P=0.005) and cholecystitis (IVW CI=1.07, 95%CI:1.02 to 1.13,P=0.010). Clostridiumsensustricto 1 was associated with an increased risk of cholelithiasis (IVW OR=1.16,95%CI:1.02 to 1.31, P=0.020) and cholecystitis (IVW OR=1.16, 95%CI:1.03 to 1.30, P=0.015). Eubacterium hallii was associated with an increased risk of primary sclerosing cholangitis (IVW OR=1.43, 95%CI: 1.03 to 1.99, P=0.033). Eubacterium ruminantium (IVW OR=0.87, 95%CI: 0.76 to 1.00, P=0.043) and Methanobrevibacter (IVW OR=0.81, 95%CI: 0.68 to 0.98, P=0.027) were associated with a reduced risk of acalculous cholecystitis. Conclusions: Eight intestinal bacterial genera maybe play pathogenic roles in benign biliary diseases. Eubacterium hallii can increase the risk of primary sclerosing cholangitis. Peptococcus and Clostridiumsensustricto 1 can increase the risk of cholelithiasis and generalized cholecystitis. Coprococcus 3 have multiple correlations with biliary stones and inflammation.

Novel mRNA isoforms and mutations of uridine monophosphate synthetase and 5-fluorouracil resistance in colorectal cancer.

The drug fluorouracil (5-FU) is a widely used antimetabolite chemotherapy in the treatment of colorectal cancer. The gene uridine monophosphate synthetase (UMPS) is thought to be primarily responsible for conversion of 5-FU to active anticancer metabolites in tumor cells. Mutation or aberrant expression of UMPS may contribute to 5-FU resistance during treatment. We undertook a characterization of UMPS mRNA isoform expression and sequence variation in 5-FU-resistant cell lines and drug-naive or -exposed primary and metastatic tumors. We observed reciprocal differential expression of two UMPS isoforms in a colorectal cancer cell line with acquired 5-FU resistance relative to the 5-FU-sensitive cell line from which it was derived. A novel isoform arising as a consequence of exon skipping was increased in abundance in resistant cells. The underlying mechanism responsible for this shift in isoform expression was determined to be a heterozygous splice site mutation acquired in the resistant cell line. We developed sequencing and expression assays to specifically detect alternative UMPS isoforms and used these to determine that UMPS was recurrently disrupted by mutations and aberrant splicing in additional 5-FU-resistant colorectal cancer cell lines and colorectal tumors. The observed mutations, aberrant splicing and downregulation of UMPS represent novel mechanisms for acquired 5-FU resistance in colorectal cancer.

Metabolism and pharmacokinetics of genipin and geniposide in rats.

Geniposide, an iridoid glucoside, is a major constituent in the fruits of Gardenia jasminoides (Gardenia fruits), a popular Chinese herb. Genipin, the aglycone of geniposide, is used to prepare blue colorants in food industry and also a crosslinking reagent for biological tissue fixation. In this study, we investigated the metabolism and pharmacokinetics of genipin and geniposide in rats. Blood samples were withdrawn via cardiopuncture and the plasma samples were assayed by HPLC method before and after hydrolysis with sulfatase and beta-glucuronidase. The results indicated that after oral administration of genipin or Gardenia fruit decoction, genipin sulfate was a major metabolite in the bloodstream, whereas the parent forms of genipin and geniposide were not detected. Importantly, oral administration of 200mg/kg of genipin resulted in a mortality of 78% (7/9) in rats.

Intervention of Online Percent Coefficient of Variation Reporting System Reduces the Variability of Tacrolimus Trough Concentration in Kidney Transplant Recipients.

BACKGROUND: The online percent coefficient of variation reporting system could monitor the variation of tacrolimus trough level (T0) and identify kidney transplant recipients (KTRs) with a higher percent coefficient of variation (%CV) instantly. Consequently, transplant doctors and pharmacists could take actions to improve drug variability. The purpose of this study was to determine the efficacy of the system for higher intrapatient variability of T0 in KTRs. METHODS: The T0 data were collected with KTRs routinely followed up at an outpatient clinic between June 2016 and November 2016. The %CV was calculated with T0 data within 6 months before and after the index date. The last outpatient clinic visit date was before December 1, 2016. The KTRs with %CV of T0 greater than 22% were enrolled. RESULTS: The study consisted of 183 KTRs (96 male, 87 female), the median age was 50 years (interquartile range [IQR], 41.0-57.0), and the median years post-kidney transplantation was 7 years (IQR, 3.0-12.4). The median T0 and creatinine level at baseline were 6.09 ng/mL (IQR, 4.80-7.52) and 1.33 mg/dL (IQR, 1.03-1.72), respectively. After the intervention, the median %CV of T0 was significantly lower than before, 32% (IQR, 26%-42%) vs 22% (IQR, 15%-33%), P < .001. The average improvement of %CV was also significantly better in KTRs with %CV ≥ 30% (median, from 41% to 25%) than KTRs with %CV between 22% and 30% (median, from 26% to 20%), P < .001. CONCLUSIONS: The results of this study indicate that continuously aggressive intervention with an online %CV reporting system effectively improves intrapatient variability of T0 in KTRs.

A comparison of the effect of carbomer-, cellulose- and mineral oil-based artificial tear formulations.

PURPOSE: To compare the efficacy, safety, and local tolerance between carbomer-based artificial tears, cellulose-, and mineral oil-based artificial tears. METHODS: A randomized, open-label, parallel-group comparative 28-day study was designed for 67 patients who were randomized into three treatment groups. Measurements included the scoring of total subjective symptoms and objective signs, Schirmer-Jones test values, and tear break-up time (BUT) at baseline, and after 2 and 4 weeks of treatment. Safety of study treatment was also assessed. Outcomes measured at baseline and 2 and 4 weeks follow-up included the scoring of total subjective symptoms and objective signs, Schirmer-Jones test values, and tear BUT, subjective assessments, and safety. RESULTS: There were no differences regarding total scores, Schirmer-Jones test, or tear BUT at baseline among these three groups at 2 and 4 weeks. Patients in all three treatment groups experienced a significant improvement from baseline in total scores and Schirmer-Jones test values after treatment. Subjective assessment was better with carbomer-based treatment. CONCLUSIONS: Each artificial tear formulation successfully relieved symptoms and signs of keratoconjunctivitis sicca. The tolerance of carbomer-based artificial tears was comparable to that of cellulose- and mineral oil-based artificial tears.

Marked decrease of cyclosporin bioavailability caused by coadministration of ginkgo and onion in rats.

Quercetin was reported to modulate CYP isoenzymes and P-glycoprotein (Pgp), a drug efflux transporter. Our previous study reported that quercetin significantly decreased the bioavailability of cyclosporin, a substrate for CYP3A4 and Pgp, in rats and pigs. Ginkgo and onion contain quercetin and its glycosides as St. John's Wort. The coadministration of cyclosporin with ginkgo or onion may be subject to clinically relevant interactions as St. John's Wort. Therefore, this study aimed to investigate the influences of ginkgo and onion on the absorption and disposition of cyclosporin in rats. Cyclosporin was administered orally and intravenously to rats with and without an oral dose of ginkgo or onion in crossover designs. Blood samples were collected via cardiopuncture and blood cyclosporin concentration was assayed by a specific monoclonal fluorescence polarization immunoassay. Everted gut sac was used to investigate the effects of ginkgo and onion on the function of intestinal Pgp. Oral coadministration of ginkgo and onion significantly decreased the Cmax of cyclosporin by 62% and 60%, and reduced the AUC0-t by 51% and 68%, respectively, whereas no influence was observed when cyclosporin was given intravenously. This indicates that the interactions between cyclosporin and ginkgo or onion occurred mainly at the absorption site. In conclusion, ginkgo and onion markedly decreased the oral bioavailability of cyclosporin. We suggest that concurrent intake of quercetin-rich herbs or foods with cyclosporin are better avoided in order to ensure the efficacy of cyclosporin.

Characterization of low-molecular-weight glutenin subunit genes from Hordeum brevisubulatum ssp. turkestanicum.

Three novel low-molecular-weight glutenin subunit (LMW-GS) genes (designated as Ht1, Ht2, and Ht3) were isolated from the genomic DNA of Hordeum brevisubulatum ssp. turkestanicum by PCR amplification (accession no. Y0695). The coding regions of Ht1, Ht2, and Ht3 were 924, 924, and 903 bp, respectively. The deduced amino acid sequences were 306, 306, and 299 amino acid residues each with a signal peptide, a central repetitive region rich in proline and glutamine, and N- and C-terminal non-repetitive domains. A comparison was carried out of these genes with other known B hordein genes from cultivated barley and LMW glutenin genes from wheat. The results indicated that Ht1, Ht2, and Ht3 had a more similar structure and a higher level of homology with the LMW-GS genes than the B hordein genes. In order to investigate the evolutionary relationship of the novel genes with the prolamin genes from barley and wheat, the phylogenetic tree was constructed and the subfamilies of these prolamin genes were identified. The results suggested that the three novel genes were glutenin-like proteins designated as LMW-m type genes.

Effects of glucose, fructose and 5-hydroxymethyl-2-furaldehyde on the presystemic metabolism and absorption of glycyrrhizin in rabbits.

Our previous study reported that co-administration of honey significantly increased the serum levels of glycyrrhetic acid (GA) after oral administration of glycyrrhizin (GZ) in rabbits. The components of honey are sucrose, glucose, fructose and 5-hydroxymethyl-furaldehyde (HMF). To clarify the causative component(s) in honey that altered the metabolic pharmacokinetics of GZ, rabbits were given GZ (150 mg kg(-1)) with and without glucose (5 g/rabbit), fructose (5 g/rabbit) and HMF (1 mg kg(-1)), respectively, in crossover designs. An HPLC method was used to determine concentrations of GZ and GA in serum as well as GA and 3-dehydroglycyrrhetic acid (3-dehydroGA) in faeces suspension. A noncompartment model was used to calculate the pharmacokinetic parameters and analysis of variance was used for statistical comparison. Our results indicated that the area under curve (AUC) of GA was significantly increased by 29% when HMF was coadministered, whereas the pharmacokinetics of GZ and GA were not significantly altered by coadministration of glucose or fructose. An in-vitro study, using faeces to incubate GZ and GA individually, indicated that HMF significantly inhibited the oxidation of GA to 3-dehydroGA and this may explain the enhanced GA absorption in-vivo. It was concluded that HMF is the causative component in honey that affects the presystemic metabolism and pharmacokinetics of GZ in-vivo.

New chemical and biological aspects of S-nitrosothiols.

Nitric oxide (NO) possesses many physiological effects and S-nitrosothiols have been identified in a variety of tissues exhibiting many NO-like activities. This review focuses on the latest discoveries pertaining to the biological functions of S-nitrosothiols and the recent research progress in the chemical properties and biomedical applications of RSNOs.

Current trends in the development of nitric oxide donors.

Nitric oxide (NO) is an important messenger molecule involved in many pathological and physiological processes within the mammalian body. Exogenous NO sources constitute a powerful way to supplement NO when the body can not generate enough for normal biological functions. In this article, general aspects on NO and NO donors are reviewed. Major focus is placed on recent developments of novel NO donors, NO releasing device(s) as well as innovative improvements to current NO donors. Finally, an outlook on future NO donor development is provided.

Shewannela putrefaciens endophthalmitis after penetrating keratoplasty.

PURPOSE: To report a case of Shewannela putrefaciens endophthalmitis after penetrating keratoplasty. METHODS: Case report. Vitreous of the recipient and the preservative medium of donor cornea were cultured. RESULTS: Vitreous of the recipient eye and the donor eye corneal preservative medium both grew S putrefaciens. The patient failed to respond to intravitreal, topical, and systemic amikacin and cefotaxime. Vision was lost rapidly. Evisceration was performed. CONCLUSION: Shewannela putrefaciens should be considered as a potential pathogen contaminating donor cornea. Shewannela putrefaciens endophthalmitis is devastating and responds poorly to medical treatment.

Profound difference in pharmacokinetics between morin and its isomer quercetin in rats.

Morin and quercetin are isomeric antioxidant flavonols widely distributed in plant foods and herbs. The pharmacokinetics of both flavonols at two doses were investigated and compared in rats. Parent forms and their glucuronides and sulfates in serum were determined by HPLC before and after enzymatic hydrolysis, respectively. After oral dosing of morin, both the parent form, morin, and its glucuronides and sulfates were present in the bloodstream. The conjugated metabolites predominated at the dose of 25 mg kg(-1), whereas the parent form was predominant at the dose of 50 mg kg(-1). Moreover, the AUC of morin parent form increased by a factor of 37 when the dose doubled, indicating that morin showed nonlinear pharmacokinetics. On the other hand, quercetin presented only as glucuronides and sulfates in the blood, indicating negligible bioavailability of quercetin, and the metabolites showed linear pharmacokinetics at the two doses studied. When considering the total AUC of parent form with conjugated metabolites, the extent of absorption of morin was 3 fold that of quercetin at the dose of 50 mg kg(-1). The results indicated that the difference in hydroxylation pattern on B-ring of flavonol markedly affected their fates in rats.

Recurrent cardiac myxoma with multiple distant metastasis and malignant change.

A 37 year-old female underwent open heart surgery for a left atrial myxoma. The post-operative course was uneventful and she was discharged two weeks later. She had regular monthly follow-up in the outpatient department until 10 months postoperatively when she was readmitted to the orthopedic ward for excision of a left ankle tumor. Two days after admission, she developed severe orthopnea. The initial diagnosis was heart failure, and she was transferred to the medical ward for treatment. Transthoracic and transesophageal echocardiography revealed a recurrent left atrial tumor. Because of acute obstruction of the mitral valve and deterioration of her condition, she underwent emergent open heart surgery. The recurrent atrial tumor was excised; histopathologic examination revealed a myxoid sarcoma. Multiple tumors were found on this admission, including a mass in the neck and in the left forearm; computed tomography revealed a brain tumor in the left posterior frontal lobe and a chest wall tumor. She died two months later. Recurrent cardiac myxoma with multiple distant metastasis may have a malignant potential. Because of the potential for tumor recurrence, long-term and regular follow-up is mandatory.

Honokiol and magnolol increased hippocampal acetylcholine release in freely-moving rats.

Honokiol and magnolol, phenolic compounds isolated from the stem bark of Magnolia officinalis, have been demonstrated to increase choline acetyltransferase activity, inhibit acetylcholinesterase, promote potassium-induced acetylcholine release and exhibit neurotrophic function in in vitro studies. The objective of the present study was to determine the effect of these compounds on hippocampal acetylcholine release in conscious, freely-moving rats. 10(-4) M-10(-6) M of honokiol or magnolol was perfused into rat hippocampus via a dialysis probe. The results showed that at 10(-4) M concentration, honokiol and magnolol markedly increased extracellular acetylcholine release to 165.5+/-5.78% and 237.83+/-9.47% of the basal level, respectively. However, lower concentrations of either compounds failed to elicit significant acetylcholine release. This result suggests that a high dose of honokiol or magnolol may enhance in vivo hippocampal acetylcholine release.

[Observations and measurements of the valves of the orbital veins].

The lumens of the superior ophthalmic vein, the intraorbital communicating branch of the angular vein and the supraorbital vein of 19 adult male cadavers (38 orbits) were studied to find lunate venous valves in 79% of the cadavers, or 68% of the orbits. Of the 71 valves found, 39% were in the communicating branch of the angular vein, 58% in the supraorbital vein, and 2.8% in the superior ophthalmic vein. The mean distances from the valves in the communicating branch of the angular vein and those in the supraorbital vein to their junction to the superior ophthalmic vein were respectively 2.12 +/- 1.46 mm and 3.05 +/- 1.84 mm. It was important to note that the valvular sinuses opened toward the cavernous sinus.

Potential modulation on P-glycoprotein and CYP3A by soymilk and miso: in vivo and ex-vivo studies.

P-glycoprotein (P-gp) and CYP3A4 both play very important roles in drug bioavailability, resistance and interactions. Our in vitro studies indicated that P-gp function was activated by many isoflavones. This study investigated the in vivo effects of soymilk and miso, isoflavone-rich soy foods, on P-gp and CYP3A by tracing the pharmacokinetics of cyclosporine (CSP), a probe drug of P-gp. Rats were orally administered CSP with and without soymilk or miso. A specific monoclonal fluorescence polarisation immunoassay was used to determine the blood concentration of CSP. The results showed that soymilk and miso significantly decreased the C(max) of CSP by 64.5% and 78.3%, and reduced the AUC(0-540) by 64.9% and 78.3%, respectively. Mechanism studies revealed that the activities of P-gp and CYP3A4 were induced by soymilk and miso. In conclusion, ingestion of soymilk and miso significantly activated the functions of P-gp and CYP3A.

3-D imaging, illustration, and quantitation of enteric glial network in transparent human colon mucosa.

BACKGROUND: Enteric glia form a network in the intestinal mucosa and have been suggested to engage in multidirectional interactions with the epithelium, blood vessels, nerves, and immune system. However, due to the dispersed nature of the glial network, standard histology cannot provide a global view of the network architecture. We prepared transparent human colon mucosa for three-dimensional (3-D) confocal microscopy with S100B immunostaining to reveal the location-dependent glial network for qualitative and quantitative analyses. METHODS: Full-thickness human colons were acquired from colectomies performed for colorectal cancer. We targeted the mucosa away from the tumor site to characterize the glial network morphology. Optical clearing (use of immersion solution to reduce scattering) was applied to generate transparent specimens for deep-tissue microscopy. KEY RESULTS: Two features of the glial network were seen: (i) A dense glial population resides at the crypt base/mucosal boundary in contact with the lymphatic vessels, and (ii) from the base, the glial network elongates along the crypt axis with peri-cryptic and peri-vascular connections toward the opening. We quantified the mucosal glia as the S100B-positive cells with at least two processes extending from the cell body. Examples of the global and in-depth imaging of adenoma were given to illustrate the morphological correlation between the loss of glial fibers and the aberrant crypts. CONCLUSIONS & INFERENCES: We have established a useful approach for 3-D imaging, panoramic illustration, and quantitation of the enteric glia in the human colon mucosa to help characterize their roles with mucosal components in health and disease.

Effect of oral azithromycin in the treatment of chlamydial conjunctivitis.

PURPOSE: To assess the efficacy of oral azithromycin in the treatment of chlamydial conjunctivitis. METHODS: We performed a retrospective study in patients with clinically suspected chlamydial conjunctivitis who underwent conjunctival swab sampling for Chlamydia direct fluorescent antibody (DFA) tests between 1 January 2006 and 31 December 2006. Patients with positive DFA results were orally administered azithromycin once a week for 2 consecutive weeks. If DFA examinations still showed positive results after 4 weeks, additional azithromycin was orally administered once. The DFA tests were repeated 4 weeks later, and this was continued until the DFA tests showed negative results. RESULTS: Among the 67 suspected patients, 45 (67.2%) showed positive results from the DFA tests, of whom 42 received treatment. After the first 2 weeks, only 27 patients returned to the clinic and completed the treatment. The test results of 19 (70.4%) patients became negative after the treatment with two weekly doses of oral azithromycin. Among the remaining eight patients, four (14.8%) needed an additional dose of oral azithromycin, and the other four (14.8%) required two additional doses. All 27 patients tolerated the treatment well, with an adverse event of mild gastritis in only one patient. CONCLUSIONS: Two weekly doses of oral azithromycin were effective and well tolerated in the treatment of chlamydial conjunctivitis. However, more than one course of treatment was necessary in some patients.

An executioner caspase regulates autophagy.

The relationships between autophagy and cell death are complex and still not well understood. To advance our understanding of the molecular connections between autophagy and apoptosis, we performed an RNAi-based screen of Drosophila melanogaster apoptosis-related genes for their ability to enhance or suppress starvation-induced autophagy. We discovered that six apoptosis-related genes, Dcp-1, hid, Bruce, buffy, debcl and p53 as well as Ras/Raf/MAPK signaling pathway components play a role in autophagy regulation in Drosophila cultured cells. Our study also provides the first in vivo evidence that the effector caspase Dcp-1 and IAP protein Bruce regulate both autophagy and starvation-induced cell death at two nutrient status checkpoints, germarium and mid-oogenesis, in the Drosophila ovary. Analysis of degenerating mid-stage egg chambers in DmAtg1 and DmAtg7 mutants reveal a reduction in TUNEL staining though DNA condensation appears unaffected. Based on these and previous findings, we propose here a putative molecular pathway that might regulate the sensitivity threshold of apoptotic and autophagic responses. We also discuss multiple interpretations of the Atg mutant egg chamber TUNEL phenotype that are consistent with a possible role for autophagy in either suppressing or enhancing the efficiency of cell degradation and/or promoting cell clearance associated with the death process.