A Novel Multiprotein Bridging Factor 1-Like Protein Induces Cyst Wall Protein Gene Expression and Cyst Differentiation in Giardia lamblia.

The capacity to synthesize a protective cyst wall is critical for infectivity of Giardia lamblia. It is of interest to know the mechanism of coordinated synthesis of three cyst wall proteins (CWPs) during encystation, a differentiation process. Multiprotein bridging factor 1 (MBF1) gene family is a group of transcription coactivators that bridge various transcription factors. They are involved in cell growth and differentiation in yeast and animals, or in stress response in fungi and plants. We asked whether Giardia has MBF1-like genes and whether their products influence gene expression. BLAST searches of the Giardia genome database identified one gene encoding a putative MBF1 protein with a helix-turn-helix domain. We found that it can specifically bind to the AT-rich initiator promoters of the encystation-induced cwp1-3 and myb2 genes. MBF1 localized to cell nuclei and cytoplasm with higher expression during encystation. In addition, overexpression of MBF1 induced cwp1-3 and myb2 gene expression and cyst generation. Mutation of the helixes in the helix-turn-helix domain reduced cwp1-3 and myb2 gene expression and cyst generation. Chromatin immunoprecipitation assays confirmed the binding of MBF1 to the promoters with its binding sites in vivo. We also found that MBF1 can interact with E2F1, Pax2, WRKY, and Myb2 transcription factors that coordinately up-regulate the cwp genes during encystation. Using a CRISPR/Cas9 system for targeted disruption of mbf1 gene, we found a downregulation of cwp1-3 and myb2 genes and decrease of cyst generation. Our results suggest that MBF1 is functionally conserved and positively regulates Giardia cyst differentiation.

A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in Giardia lamblia.

Giardia lamblia persists in a dormant state with a protective cyst wall for transmission. It is incompletely known how three cyst wall proteins (CWPs) are coordinately synthesized during encystation. Meiotic recombination is required for sexual reproduction in animals, fungi, and plants. It is initiated by formation of double-stranded breaks by a topoisomerase-like Spo11. It has been shown that exchange of genetic material in the fused nuclei occurs during Giardia encystation, suggesting parasexual recombination processes of this protozoan. Giardia possesses an evolutionarily conserved Spo11 with typical domains for cleavage reaction and an upregulated expression pattern during encystation. In this study, we asked whether Spo11 can activate encystation process, like other topoisomerases we previously characterized. We found that Spo11 was capable of binding to both single-stranded and double-stranded DNA in vitro and that it could also bind to the cwp promoters in vivo as accessed in chromatin immunoprecipitation assays. Spo11 interacted with WRKY and MYB2 (named from myeloblastosis), transcription factors that can activate cwp gene expression during encystation. Interestingly, overexpression of Spo11 resulted in increased expression of cwp1-3 and myb2 genes and cyst formation. Mutation of the Tyr residue for the active site or two conserved residues corresponding to key DNA-binding residues for Arabidopsis Spo11 reduced the levels of cwp1-3 and myb2 gene expression and cyst formation. Targeted disruption of spo11 gene with CRISPR/Cas9 system led to a significant decrease in cwp1-3 and myb2 gene expression and cyst number. Our results suggest that Spo11 acts as a positive regulator for Giardia differentiation into cyst.

New Mitogenomes of Two Chinese Stag Beetles (Coleoptera, Lucanidae) and Their Implications for Systematics.

Although conspicuous and well-studied, stag beetles have been slow to join the genomic era. In this study, mitochondrial genomes of two stag beetles, Sinodendron yunnanense and Prosopocoilus confucius, are sequenced for the first time. Both of their genomes consisted of 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNAs (rRNAs), and a control region. The mitogenome of S. yunnanense was 16,921 bp in length, and P. confucius was 16,951 bp. The location of the gene trnL(UUR), between the A + T-rich and control region in S. yunnanense, is the first observed in Lucanidae. In P. confucius, an unexpected noncoding region of 580 bp was discovered. Maximum likelihood and Bayesian inference on the 13 mitochondrial PCGs were used to infer the phylogenetic relationships among 12 representative stag beetles and three scarab beetles. The topology of the two phylogenetic trees was almost identical: S. yunnanense was recovered as the most basal Lucanid, and the genus Prosopocoilus was polyphyletic due to P. gracilis being recovered sister to the genera Dorcus and Hemisodorcus. The phylogenetic results, genetic distances and mitogenomic characteristics call into question the cohesion of the genus Prosopocoilus. The genetic resources and findings herein attempts to redress understudied systematics and mitogenomics of the stag beetles.

Palladium-catalyzed enantioselective C-H arylation for the synthesis of P-stereogenic compounds.

A palladium-catalyzed enantioselective C-H arylation of N-(o-bromoaryl)-diarylphosphinic amides is described for the synthesis of phosphorus compounds bearing a P-stereogenic center. The method provides good enantioselectivities and high yields. The products were readily transformed into P-chiral biphenyl monophosphine ligands.

[Endothelial protection of tanshinone in rats of severe acute pancreatitis].

OBJECTIVE: To investigate the protective effection of tanshinone on endothelial cells of severe acute pancreatitis (SAP) rats and the effection of tanshinone on apoptosis of aorta endothelium. METHODS: Using 8% L-arginine intraperitoneal to inject in rats, 4.4 mg/g per time, repeat injection 1 hour later, for establishing SAP model. Model rats were randomly divided into SAP group and tanshinone group. 20 mg/kg Sodium Tanshinon II Asilate i. p. was applied to tanshinone group,while the saline was used to replace Sodium Tanshinon II Asilate in SAP group. Twelve rats of each group were sacrificed at 12 h, 24 h after treatment. The pathological changes in pancreatic tissues were observed. Abdominal aorta samples were collected for terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labelling (TUNEL) and reverse transcription PCR (RT-PCR) tests. The blood samples were collected from abdominal aorta for analysis. Detections: (1) The concentration of Von Willebrand factor (vWF), soluble endothelial protein C receptor (sEPCR), tumor necrosis factor alpha (TNF-alpha) and the serum levels of nitric oxide (NO) were quantitative messured by ELISA. (2) The apoptosis of aorta endothelium cell was examined using TUNEL method. (3) Bcl-2 and Bax mRNA expression were measured by RT-PCR. RESULTS: The pathological changes of pancreatic tissues were more severe in SAP group than those in tanshinone group. Compared with SAP group, treatment with tanshinone effectively inhibited TNF-alpha (P < 0.05), vWF (P < 0.05) and sEPCR (P < 0.05) expression and depressed apoptosis of aorta endothelium cell, increased the expression of Bcl-2 mRNA (P < 0.05), Bcl-2 mRNA/Bar mRNA ratio (P < 0.05) and the expression of Bax mRNA (P < 0.05) were decreased significantly. CONCLUSION: Sodium Tanshinon II Asilate can lighten the SAP rats aortic endothelial injury and apoptosis of endothelial cells can reduce endothelial damage of SAP rats by TNF-alpha expression suppression.

Human leukocyte antigen-DR expression on peripheral monocytes may be an early marker for secondary infection in severe acute pancreatitis.

BACKGROUND/AIMS: To investigate whether the human leukocyte antigen-DR (HLA-DR) expression on peripheral monocytes can be utilized as a precursor to a secondary infection of severe acute pancreatitis (SAP). METHODOLOGY: Patients diagnosed with SAP who were admitted into West China Hospital within 48 h after symptom onset from July 1, 2010 to December 31, 2010 (n = 40) were included. HLD-DR expression on peripheral monocytes on the 1st, 3rd, 5th and 7th day of hospitalization was detected with flow cytometry analysis to determine whether a prediction could be made in regards to development of a secondary infection. RESULTS: There were 11 patients with secondary infection complications, 4 of which died during hospitalization. On the 1st, 3rd, 5th and 7th day, HLA-DR expression on monocytes in the infected patients was lower than those in the noninfected patients (P < 0.05). There was no statistical significance in the serum CRP and APACHE II between the groups on the first day (P > 0.05). Upon initial admission HLA-DR expression showed a negative correlation with longer-term admission APACHE II (r = -0.790, P = 0.000) and serum CRP (r = -0.642, P = 0.000). The area under the ROC curve (AUC) was 0.837 (95%CI: 0.685-0.989, P = 0.001) for admission HLA-DR, 0.809 (95% CI: 0.667-0.951; P = 0.003) for APACHE II score and 0.781 for serum CRP (95% CI: 0.627-0.934; P = 0.007) to predict secondary infection. The cut-off value of prediction of secondary infection was 35.8% in HLA-DR expression with a sensitivity of 81.8% and a specificity of 82.8%, 10.5 in APACHE II on admission with a sensitivity of 90.9% and a specificity of 48.3%, 155 mg/L in serum CRP on admission with a sensitivity of 90.9% and a specificity of 44.8%. CONCLUSIONS: The HLA-DR expression on monocytes may be an ideal marker for an early prediction of secondary infection in SAP.

[Clinical study on different admission serum glucose levels in patients with severe acute pancreatitis].

OBJECTIVE: To investigate the clinical characteristics, prognosis effects and management of different admission serum glucose levels in patients with severe acute pancreatitis (SAP). METHODS: A retrospective analysis involving a total of 218 patients with SAP and have serum glucose > or = 6.1 mmol/L at admission during the period from August 1, 2005 to December 31, 2007 was enrolled based on the coding data in West China hospital. They were divided into 3 groups according to admission glucose levels of 6.1-11.1 mmol/L (n = 115), 11.2-16.7 mmol/L (n = 71), and > 16.7 mmol/L (n = 32) respectively. Patients' demographic characteristics, clinical parameters, various scoring systems, the ICU transfer rate during early phase and the mortality, infection rate and operation transfer rate during later phase were obtained and calculated. RESULTS: The pulse and respire frequency, the levels of serum lactate dehydrogenase (LDH), blood urea nitrogen (BUN), creatinine (Crea) and scores on the Ranson Criteria, Acute Physiology and Chronic Health Evaluation II (APACHE II) at admission, APACHE II and Balthazar's Computed Tomography Severity Index (CTSI) within 72 hours increased in sequence according to mild, moderate and severe hyperglycemic group (P < 0.01). Whereas the concentration of serum Ca2+ was lower than that in the mild elevated serum glucose group (P < 0.01). In the 3 groups, the early single organ failure rates were elevated in turn (P < 0.01) and the multi-organ failure rates were 16.5%, 45.1% and 50.0% (P < 0.01) respectively. Simultaneously, the ICU transfer rates were 10.4%, 26.8% and 34.4%, while the mortality in the 3 groups were 7.8%, 16.9% and 40.6% respectively, which were statistically significant (P < 0.01). CONCLUSION: The findings of the study suggest that admission elevated glucose is an indicator of organ failure and poor prognosis of SAP.

[Health economic evaluation of clinical pathway of traditional Chinese medicine in the treatment of mild acute pancreatitis].

OBJECTIVE: To evaluate the health economic value clinical pathway (CP) of traditional Chinese medicine in the treatment of mild acute pancreatitis (MAP). METHODS: Ninty one patients with MAP were enrolled prospectively in TCM clinical pathway group from June 2012 to February 2013, while the data of 80 MAP patients who were treated without TCM clinical pathway from June 2011 to May 2012, were analyzed retrospectively as control group. The health economic evaluation data used for the two groups comparison included: average length of stay, hospitalization expenses (total hospitalization expenses, total treatment cost, TCM treatment cost, herbal fees, medicine fees, and nursing care cost), as well as the usage of antibiotics/somatostatin, the release time of abdominal pain, the time of re-feeding, and patient satisfaction. RESULTS: There were no significant statistical differences in demographics, etiology, Ranson and Balthazar CT scores between the two groups (P > 0.05). Compared with non-CP group, the usage of antibiotics and somatostatin, the release time of abdominal pain, the time of re-feeding and patient satisfaction were all improved significantly in CP group (P < 0.05). The average length of stay in CP group was shorter than that of non-CP group (P < 0.05). Total hospitalization expenses [yen (11,089.89 +/- 4,318.29) vs. yen (8,960.34 +/- 4,328.91)], medicine fees [yen (6,563.80 +/- 2,743.87) vs. yen (3,988.28 +/- 2,128.10)] and nursing care cost [yen (110.51 +/- 37.24) vs. yen (93.32 +/- 35.20)] were all reduced in CP group, while TCM treatment cost [yen (609.59 +/- 624.42) vs. (968.29 +/- 769.68)] and herbal fees [yen (162.72 +/- 135.13) vs. yen (303.49 +/- 149.90)] were increased (P < 0.05). There was no significant statistical difference in total treatment cost between the two groups (P > 0.05). CONCLUSION: TCM clinical pathway of MAP can not only ensure the therapeutic effects, but also shorten the average length of stay, reduce medical cost and increase patient satisfaction.

[The value of different scoring systems for computed tomography in predicting pancreatic pseudocyst in severe acute pancreatitis].

OBJECTIVE: To evaluate the value of several Computed Tomograph (CT) scoring systems in predicting the development of acute pancreatic pseudocyst (PPC) in severe acute pancreatitis (SAP) during early One hundred and sixty-two patients with SAP were retrospectively observed and subjected to stage. METHODS clinical, laboratory, and radiology investigation from October 2007 to December 2010. Three different CT scoring systems including CT severity index (CTSI), Modified CT severity index (MCTSI) and Extrapancreatic Inflammation on CT score (EPIC), were used for the determine of PPC, while the predictive values of these three Forty-eight patients CT scoring systems in the presence of PPC were analyzed by the ROC curve. RESULTS: (29.6%) were observed the formation of PPC. The scores of CTSI, MCTSI, EPIC and the occurrence rate of ascites in PPC group were significantly higher than those in non-PPC group with One-way ANOVA analysis. Among the three CT scoring systems, EPIC score showed a larger area under ROC curve (AUC = 0.914) than CTSI (AUC = 0.674) and MCTSI (AUC = 0.72) did. CONCLUSION: EPIC scoring system has better prediction of PPC in SAP patients than CTSI and MCTSI.

Vitamin D and Pancreatitis: A Narrative Review of Current Evidence.

Emerging research indicates that vitamin D metabolic disorder plays a major role in both acute pancreatitis (AP) and chronic pancreatitis (CP). This has been demonstrated by studies showing that vitamin D deficiency is associated with pancreatitis and its anti-inflammatory and anti-fibrotic effects by binding with the vitamin D receptor (VDR). However, the role of vitamin D assessment and its management in pancreatitis remains poorly understood. In this narrative review, we discuss the recent advances in our understanding of the molecular mechanisms involved in vitamin D/VDR signaling in pancreatic cells; the evidence from observational studies and clinical trials that demonstrate the connection among vitamin D, pancreatitis and pancreatitis-related complications; and the route of administration of vitamin D supplementation in clinical practice. Although further research is still required to establish the protective role of vitamin D and its application in disease, evaluation of vitamin D levels and its supplementation should be important strategies for pancreatitis management according to currently available evidence.