Horseshoe kidney with PLA2R-positive membranous nephropathy.

BACKGROUND: Horseshoe kidney (HSK) is a common congenital defect of the urinary system. The most common complications are urinary tract infection, urinary stones, and hydronephrosis. HSK can be combined with glomerular diseases, but the diagnosis rate of renal biopsy is low due to structural abnormalities. There are only a few reports on HSK with glomerular disease. Here, we have reported a case of PLA2R-positive membranous nephropathy occurring in a patient with HSK. CASE PRESENTATION: After admission to the hospital due to oedema of both the lower extremities, the patient was diagnosed with nephrotic syndrome due to abnormal 24-h urine protein (7540 mg) and blood albumin (25 g/L) levels. Abdominal ultrasonography revealed HSK. The patient's brother had a history of end-stage renal disease due to nephrotic syndrome. Therefore, the patient was diagnosed with PLA2R-positive stage II membranous nephropathy through renal biopsy under abdominal ultrasonography guidance. He was administered adequate prednisone and cyclophosphamide, and after 6 months of treatment, urinary protein excretion levels significantly decreased. CONCLUSION: The risk and difficulty of renal biopsy in patients with HSK are increased due to structural abnormalities; however, renal biopsy can be accomplished through precise positioning with abdominal ultrasonography. In the literature, 20 cases of HSK with glomerular disease have been reported thus far. Because of the small number of cases, estimating the incidence rate of glomerular diseases in HSK is impossible, and the correlation between HSK and renal pathology cannot be stated. Further studies should be conducted and cases should be accumulated to elucidate this phenomenon.

[Effects of Emodin Derivative on Cell Cycle, Apoptosis and NF-κB Pathway in Burkitt Lymphoma Cells].

OBJECTIVE: To investigate the effect and mechanism of a novel emodin derivative YX-18 on Burkitt lymphoma (BL) cells. METHODS: MTT assay was used to detect the effect of YX-18 on the proliferation of BL cell lines CA46 and Raji. Annexin V-PE/7-AAD double staining assay was used for detecting the effect of YX-18 on the apoptosis of CA46 and Raji cells. PI/RNase staining was used to test the effect of YX-18 on CA46 and Raji cell cycle. JC-1 method was used to measure the changes of mitochondrial membrane potential after YX-18 treatment, and DAPI staining was used to detect the morphology of apoptotic cells. Western blot was used to analyze the distribution changes of NF-κB pathway protein (P65, P-P65, IκB, P-IκB) in the cytoplasm and cell nucleus, and also the expression changes of cyclin-related protein P21, CDK2, P-CDK2, Cycling D1, Cycling E1, and the apoptosis-related protein Caspase-3, Caspase-8, Caspase-9 and the proliferation-related protein C-MYC, BCL-2 by YX-18. Real-time fluorescence-quantitative PCR was used to evaluate the effects of YX-18 on mRNA levels of C-MYC and Ki-67 genes in CA46 and Raji cells, and EBNA-1 and EBER genes of EBV in Raji (EBV+) cells. RESULTS: Novel Emodin derivative YX-18 could effectively inhibit the proliferation of BL cell lines CA46 and Raji, showing a time-dependent effect (24, 48 and 72 h: rCA46=0.89, 0.75, 0.75, rRaji=0.87, 0.73, 0.64). IC50 of CA46 cells and Raji cells treated with YX-18 for 24 h was 1.77±0.04 µmol/L and 1.97±0.22µmol/L, respectively. CA46 cells and Raji cells were treated with YX-18 at concentration of 2.0 and 4.0 µmol/L for 24 h. Compared with the control group, both strains of cells showed a very significant apoptosis at the concentration of 2.0 and 4.0 µmol/L (P<0.01), showing a concentration-dependent effect (rCA46=0.99, rRaji=0.92). Moreover, the cleavaged Caspase-3, 8 and 9 proteins were activated by YX-18 into verious degrees in both two cell lines. Both the two cell lines displayed by YX-18 cell cycle arrest at G0/G1 phase (P<0.01) after exposed to YX-18 for 24 hours at the concentration of 1.0, 2.0 µmol/L in CA46 cells and at 0.5 and 1 µmol/L in Raji cells, respectively. YX-18 decreased expression level of cyclin D1, cyclin E1, CDK2, p-cdk2 proteins and increased p21Waf1/Cip1 level in CA46 and Raji cells. YX-18 significantly declined mitochondrial membrane potential in both cells at the concentration of 2.0 and 4.0 µmol/l (P<0.01) with concentration-dependent manner (rCA46=-0.96, rRaji=-0.99). Western blot tests indicated that YX-18 down-regulated nucleus P65 and intracellular cytoplasm P65, P-IκB, P-P65 protein, and upregulated intracellular IκB level with dose-dependent manner. Meanwhile, the expression level of the cell proliferation-related molecules C-MYC and BCL-2 was decreased significantly. YX-18 suppressed mRNA levels of C-MYC and Ki-67 in both cell lines, and EBNA-1 in EBV-positive Raji cells in a concentration-dependent way. CONCLUSION: The novel emodin derivative YX-18 can significantly inhibit the proliferation of Burkitt lymphoma cells, and induce the cell apoptosis and cycle arrest. The inhibitory effect of YX-18 on the proliferation of Burkitt lymphoma cells may be related with the effect of Caspase apoptosis pathway, the proliferation and apoptosis-related molecules, such as C-MYC and Ki-67, and also to the inhibition of NF-κB pathway.

Positive endoscopic and ultrasonographic findings in patients with symptom-diagnosed functional gastrointestinal disorder: Data from a Chinese cross-sectional study.

OBJECTIVE: Few studies have systematically assessed symptom-based diagnoses of functional gastrointestinal disorders (FGID) in China. This study aimed to evaluate the positive endoscopic and ultrasonographic findings in patients with symptom-based FGID. METHODS: Adult patients with gastrointestinal symptoms but not red flag symptoms who were empirically diagnosed with FGID underwent an upper or lower gastrointestinal endoscopy. An abdominal ultrasonography was also performed to screen for potential organic diseases. Patients' data were collected and the Rome III questionnaire was used for symptom-based assessment. RESULTS: Positive endoscopic and ultrasonographic findings were detected in 31.8%, 26.2% and 9.9% of patients with esophageal, gastroduodenal and intestinal disorders, respectively. Among patients diagnosed with FGID using the questionnaire, the rates of positive endoscopic and ultrasonographic findings were higher in patients with upper gastrointestinal symptoms than in those with lower gastrointestinal symptoms. Moreover, patients elder than 34.5 years with upper gastrointestinal symptoms and those elder than 47.5 years with lower gastrointestinal symptoms were more likely to have positive endoscopic and ultrasonographic findings. CONCLUSIONS: The Rome III questionnaire is a valuable diagnostic tool for screening FGID in China. However, the rates of positive endoscopic and ultrasonographic findings vary depending on the location of symptoms. Endoscopy and ultrasonography have irreplaceable value, especially for investigating upper gastrointestinal symptoms.

[Inhibitory and Inducing Effects of Emodin Derivative E11 on Proliferation and Apoptosis of Multiple Myeloma Cells].

OBJECTIVE: To screen the most strong emodin derivative inhibiting the proliferation of multiple myeloma(MM) cells and to explore the inhibitory and inducing effects of emodin derivatives on proliferation and apoptosis of MM cell lines RPMI 8226 and U266. METHODS: Sixteen emodin derivatives were designed and synthesized by using emodin as mother substance, then from which the emodin derivative E11 was screened for experiments. The MTT method and cell colony formation assay were used to observe the effect of E11 on proliferation of RPMI 8226 and U266, the fluorescent microscopy with DAFI staining was used to observed the morphological changes of MM cells treated with emodin dervative 11, the DNA fragmentation detection was used to detect the inducing apoptosis effect of E11 on RPMI 8226 and U266 cells treated with E11. RESULTS: The MTT assay showed that after the RPMI 8226 cells were treated with 16 kinds of emodin derivatives for 48 hours, the 50% inhibition concentration（IC50） of 14 emodin dervatives was between 0.83-34.68 µmol/L, except E10 and E15 because their IC50 could not be calculated. The IC50 of E11 for RPMI 8226 and U266 cells were 0.831±0.0453 µmol/L and 1.039±0.093 µmol/L, respectively. Cell colony formation assay showed that E11 could inhibit RPMI8226 and U266 cells' colony formation in dose-.and time- dependent manner (r=0.72). Cell apoptosis was observed in RPMI8226 and U266 cells by DAPI staining , and also by the detection of DNA fragmentation. CONCLUSION: In the synthesis of 16 kinds of emodin derivatives, the inhibitory effect of E11 on prolife-ration of RPMI8226 cell was the strongest. E11 can remarkably inhibit proliferation and induce apoptosis of RPMI8226 and U266 cells.

[Effect of A Novel Emodin Derivative on Chronic Myelogenous Leukemia K562 Cells and Imatinib-resistant K562/G01 Cells].

OBJECTIVE: To explore the effect of a novel emodin derivative E19 on proliferation inhibition and apoptosis induction of human chronic myelogenous leukemia (CML) cell line K562 and imatinib-resistant CML cell line (K562/G01), and to clarify the involved mechanisms. METHODS: MTT and colony formation test were used to detect the cell proliferation. Apoptotic induction effects were examined by DAPI staining method and DNA ladder assay. Western blot was performed to detect the changes of P210(Bcr-Abl) protein. RESULTS: The emodin derivative E19 could efficiently inhibit proliferation and induce apoptosis in K562 and K562/G01 cells. IC50 of K562 cells and IC50 of K562/G01 cells were (1.20 ± 0.19) µmol/L and (1.22 ± 0.16) µmol/L, respectively. DNA fragmentation in K562 cells and K562/G01 cells confirmed that the E19 induced apoptosis in dose-dependent manner. Western blot showed that emodin derivative inhibited phosphorylation of P210 protein in K562 cells and K562/G01 cells and down-regulated the expression level of P210 in dose- and time-dependent manners. CONCLUSION: The emodin derivative E19 can efficiently inhibit growth and induce apoptosis of K562 cells and K562/G01 cells, while the inhibition of phosphorylation of P210 protein and down-regulation of P210 protein expression may be involved in these processes.

[Effect of Emodin Derivative E11 on T Lymphocytic Leukemia Cell Line Molt-4 and Its Possible Mechanisms].

OBJECTIVE: To explore the effect of a new emodin derivative E11 on proliferation and apoptosis of T lymphocytic leukemia cell line Molt-4 and its possible mechanisms. METHODS: MTT method was used to plot cell growth curve. Colony culture assay was performed for studying the effect of emodin derivative E11 on colony-formation of Molt-4. The fluorescent microscopy with DAPI staining was used to examine the cell morphological changes after E11 treatment. DNA fragmentation method was used to detect the inducing effect of emodin derivative E11 on cell apoptosis. Western blot was used to determine the expressions of apoptosis-related proteins including procaspase-9, procaspase-3, PARP and PI3K/AKT, MAPK signalling pathway. RESULTS: Emodin derivative E11 could strongly inhibit the growth of Molt-4 with the IC50 in 48 h at 1.381 ± 0.1552 µmol/L in dose-dependent manner. 0.1 µmol/L of E11 could inhibit cell colony formation. The typrical apopototic morphologic changes of Molt cells treated with E11 could be observed under fluorescence microscope with DAPI staining. DNA apoptotic ladder could be observed by DNA fragmentation.The expressions of procaspase -9, procaspase-3, PARP, p-MAPK, p-AKT, mTOR, p-mTOR, p-P70 and p-4BEP1 were down-regulated, while expressions of MAPK, AKT, 4EBP1 and P70 were not changed remarkably after Molt-4 were treated with E11 for 48 h. CONCLUSION: E11 can remarkably inhibit the proliferation and induce the apoptosis of Molt-4 cells. The mechanism of apoptosis of Molt-4 cells may be related with the suppression of PI3K/AKT and MAPK signalling pathways.

Nationwide prevalence of human papillomavirus infection and viral genotype distribution in 37 cities in China.

BACKGROUND: Type-specific high-risk HPV (hrHPV) infection is related to cervical carcinogenesis. The prevalence of hrHPV infection varies geographically, which might reflect the epidemiological characteristics of cervical cancer among different populations. To establish a foundation for HPV-based screening and vaccination programs in China, we investigated the most recent HPV prevalence and genotypic distributions in different female age groups and geographical regions in China. METHODS: In 2012, a total of 120,772 liquid-based cytological samples from women enrolled for population- or employee-based cervical screening in 37 Chinese cities were obtained by the Laboratory of Molecular Infectious Diseases of Guangzhou KingMed. A total of 111,131 samples were tested by Hybrid Capture II and the other 9,641 were genotyped using the Tellgenplex™ HPV DNA Assay. RESULTS: The total positive rate for hrHPV was 21.07 %, which ranged from 18.42 % (Nanchang) to 31.94 % (Haikou) and varied by region. The regions of Nanchang, Changsha, Hangzhou, Chengdu, Fuzhou, Guangdong, and Guiyang could be considered the low prevalence regions. Age-specific prevalence showed a "two-peak" pattern, with the youngest age group (15-19 years) presenting the highest hrHPV infection rate (30.55 %), followed by a second peak for the 50-60-year-old group. Overall, the most prevalent genotypes were HPV16 (4.82 %) and HPV52 (4.52 %), followed by HPV58 (2.74 %). Two genotypes HPV6 (4.01 %) and HPV11 (2.29 %) were predominant in the low-risk HPV (lrHPV) type, while the mixed genotypes HPV16 + 52 and HPV52 + 58 were most common in women with multiple infections. CONCLUSIONS: This study shows that HPV infection in China has increased to the level of an "HPV-heavy-burden" zone in certain regions, with prevalence varying significantly among different ages and regions. Data from this study represent the most current survey of the nationwide prevalence of HPV infection in China, and can serve as valuable reference to guide nationwide cervical cancer screening and HPV vaccination programs.

Chemical and material communication between the optic nerves in rats.

BACKGROUND: To examine interactions between optic nerves. METHODS: A total of 24 Sprague-Dawley rats received unilateral intravitreal injections. The rats were equally divided into four groups: group A was administered an adeno-associated virus (AAV) carrying an exogenous gene (ND4; rAAV-ND4); group B, AAV carrying a green fluorescent protein (GFP; rAAV-GFP); group C, fluorogold (FG) nerve tracer dye; and group D, phosphate-buffered saline (PBS) as a control. Two weeks later, GFP expression was evaluated in both retinas and optic nerves of group B rats after frozen sectioning. The presence of FG was also evaluated in group C optic nerves by fluorescent microscopy after frozen sectioning. Four weeks after injection, ND4 expression was evaluated in both eyes of groups A and D using western blotting and immunofluorescence. RESULTS: FG was observed in the optic chiasm posterior segment along the optic nerve of injected eyes. Some FG reached the anterior optic nerve of the non-injected eye. GFP fluorescence was observed only in the retina of the injected eye but not in the contralateral retina or either optic nerve. ND4 expression was significantly different between injected and non-injected eyes but not between the non-injected eyes in groups A and D. CONCLUSION: Unilaterally injected material can reach the contralateral optic nerve through axoplasmic transport. It is possible that this the only mechanism by which the optic nerves directly communicate.

[Corrigendum] Overexpression of the chimeric plasmin-resistant VEGF165/VEGF183 (132-158) protein in murine breast cancer induces distinct vascular patterning adjacent to tumors and retarded tumor growth.

After the publication of the article, the authors noted that they had made an error regarding certain facts in their manuscript: In the abstract VEGF192 (132-158) should be changed to VEGF183 (132-158) (Page 1, Line 2). In addition, width should be changed to width2 (Page 3, Line 50). The authors regret these errors. [the original article was published in the Molecular Medicine Reports 11: 1483-1489, 2015 DOI: 10.3892/mmr.2014.2866]

Overexpression of the chimeric plasmin-resistant VEGF165/VEGF183 (132-158) protein in murine breast cancer induces distinct vascular patterning adjacent to tumors and retarded tumor growth.

A chimeric plasmin­resistant vascular endothelial growth factor (VEGF)165/VEGF183 (132-158) protein, named as VEGF183 (according to the nomenclature of VEGF), designed by a previous study, was demonstrated to have an enhanced affinity for the extracellular matrix (ECM) amongst other bioactivities. However, it is now accepted that mutant VEGFs frequently demonstrate different angiogenic activities and produce different vascular patterning from the parental molecule. The present study hypothesized that VEGF183, due to its enhanced binding affinity to the ECM, would exhibit a different angiogenic activity and produce a different vascular patterning compared to those of VEGF165. Murine breast cancer EMT­6 cells were manipulated to stably overexpress VEGF165 or VEGF183. These cells were then inoculated intradermally into BALB/c mice in order to monitor the formation of vascular patterning in skin proximal to tumors. In vivo angiogenesis experiments revealed that overexpression of VEGF183 in murine breast cancer cells resulted in irregular, disorganized and dense vascular patterning as well as induced a significant inhibition of tumor growth compared with that of VEGF165. In addition, allograft tumor immunochemical assays of VEGF183­overexpressing tumors demonstrated significantly lower vascular densities than those of VEGF165­overexpressing tumors; however, VEGF183 tumors had a significantly enlarged vascular caliber. Conversely, cell wound healing experiments revealed that VEGF183­overexpressing EMT­6 cells had significantly decreased migration rates compared with those of VEGF165­overexpressing EMT­6 cells. In conclusion, the results of the present study supported the hypothesis that the altered ECM affinity of VEGF induced structural alterations to vasculature. In addition, these results provided a novel insight into VEGF design and indirect evidence for the function of exon 8 in VEGF. [Corrected]

[Synthesis of emodin derivatives and their inhibiting effects on proliferation of leukemia cell lines].

The aim of this study was to explore the inhibitory effect of newly synthesised emodin derivatives on the proliferation of leukemia cell lines and to select the most effective one from these emodin derivatives for further research. Emodin derivatives were synthesized by modifying the structure of emodin. MTT method was used to detect the proliferative inhibition in leukemia cell lines treated with emodin derivatives. The results showed that the half inhibitory concentration (IC50) for K562 cells treated with emodin derivatives E10-19 for 48 h was 0.84 - 12.01 µmol/L. E19 displayed the best anti-proliferative activity, while E16 and E17 did not show effects on K562 cells. Emodin derivative E19 was chosen for treating U937, NB4, Molt-4 and CA-46 cells, their IC50 for 48 h were 0.85, 0.9, 0.76, 0.8 µmol/L respectively. The IC50 of E19 for LQ2 cells was 3.60 µmol/L, and the IC50 range of E19 for normal human peripheral blood mononuclear cells at 48 h was 4.01 - 4.78 µmol/L. It is concluded that emodin derivative E19 can strongly inhibit the growth of leukemia cells and its inhibiting effect on proliferation of leukemia cells has a certain specificity. The specific mechanism of E19 anti-leukemia effect should be further studied.

A novel effect of imidazole derivative KK-42 on increasing survival of Aeromonas hydrophila challenged prawn Macrobrachium nipponense.

Imidazole derivative KK-42 is well known as the insect growth regulator. Here we find that KK-42 pretreatment could promote the survival of Macrobrachium nipponense infected with Aeromonas hydrophila, which is considered to be possibly related to the prophenoloxidase (proPO), a conserved copper-containing enzyme that plays an important role in defense against pathogens. In this study, a full-length of proPO gene from M. nipponense haemocytes, designated as MnproPO, was firstly cloned and characterized. The full-length cDNA contained 2428 bp with a 2013 bp open reading frame encoding a putative proPO protein of 671 amino acids with a predicted molecular mass of 76.5 kDa and pI of 7.31. It was predicted to possess all the expected features of proPO members, including two putative copper-binding sites with six histidine residues and a thiol ester-like motif. Sequence analysis showed that MnproPO exhibited the highest amino acid sequence similarity (93%) to a proPO of Macrobrachium rosenbergii. The gene was expressed highly in haemocytes and weakly in hepatopancreas. Real-time PCR analysis revealed that the MnproPO expression increased significantly at 3, 12 and 24 h after KK-42 treatment, the PO activity also importantly rose from 6 to 48 h in KK-42-treated prawns and reached the maximum at 24 h with a 2.3-fold higher than that in control group. Injection of A. hydrophila could stimulate the MnproPO transcription and PO activity whether or not the prawns were pretreated by KK-42, the mRNA level increased obviously only at 3 h and 6 h after the bacterium injection (challenged control), but increased constantly during the phase of experiment except at 6 h under the condition of KK-42 pretreatment (challenged treatment group). The change trend of PO activity was basically similar to that of MnproPO expression. Our present results demonstrate that the MnproPO expression as well as PO activity may be induced by KK-42, which is likely one of the molecular mechanisms of KK-42 acts for increasing survival of the prawn infected with A. hydrophila.

[Phenotypic analysis of Th cells in colon and peripheral blood in patients with irritable bowel syndrome].

UNLABELLED: OBJECTIVE; To analyze the change of Th1/Th2/Th17 in colonic mucosa and peripheral blood in diarrhea-predominant IBS (D-IBS) to uncover the underlying mechanism for the activation of mucosal immune system. METHODS: Colonic biopsy specimens and peripheral blood were obtained from patients with D-IBS (n = 27) and controls (n = 16). Two different groups were classified on the basis of histological assessment of biopsy specimens from D-IBS patients. One group (14 of 27) had normal conventional histology (IBS), while another group (13 of 27) had nonspecific microscopic inflammation (IBS-A). Flow cytometric detection of intracellular IFN-gamma/IL-4/IL-17 cytokine production was employed to investigate Th1, Th2 and Th17 cells in colonic lamina propria and peripheral blood. Western blot was used to determine the expressions of IL-12, IL-4 and IL-17 in colonic mucosa. The levels of IL-12, IL-4 and IL-17 in peripheral blood were detected by ELISA. RESULTS: In colonic mucosa, the proportion of Th17 increased in IBS-A group as compared with controls [3.60 (4.05) vs 1.25 (3.70), P = 0.045], but not in IBS group. No difference could be observed in the frequencies of Th1 and Th2 in colonic mucosa and peripheral blood. The levels of IL-12, IL4 and IL-17 in IBS and IBS-A showed no difference in either colonic mucosa or peripheral blood. CONCLUSION: Subgroup of D-IBS showed abnormal conventional histology, implicating the activation of mucosal immune system in pathogenesis. The shift of Th1/Th2/Th17 balance in colonic mucosa showed the enhanced Th17 activity.

[The triplet properties of beta-carotene in acetonitrile solution: a laser flash photolysis study].

The representative of carotenoids, beta-carotene, can scavenge reactive oxygen radicals like singlet molecular oxygen, nitrogen dioxide radical and peroxyl radical due to the effective antioxidative properties. In medicine, beta-carotene is used to alleviate the disease erythropoietic protoporphyria (EPP), by intercepting the triplet state of protoporphyrin (a porphyrin lacking a central metalion, a precursor to haem) therefore preventing the formation of singlet oxygen. Epidemiological evidence has suggested that dietary beta-carotene may inhibit certain types of cancer. Much of work has been carried out in benzene, toluene, or chloroform as most caroienoids are sufficiently soluble in these nonpolarity solvents. In the present paper, the generation and properties of triplet beta-carotene in acetonitrile solution were investigated with 355 nm laser flash photolysis. 2-acetonaphthone was used as an excitation energy donor to sensitize the production of the triplet state of beta-arotene. Excitation of the solution containing 2-acetonaphthone and beta-carotene upon 355 nm laser flash produced the triplet of 2-acetonaphthone (420 nm) firstly. Subsequently, the excitation energy of triplet 2-acetonaphthone was transferred to beta-carotene generating triplet beta-arotene. Characteristic absorption spectra of triplet beta-arotene (510 nm) were recorded. By means of transfer of excitation energy, the molar absorption coefficients of triplet beta-arotene were determined to be 23 000 dm3 mol(-1) x cm(-1) at 510 nm. The triplet lifetime for beta-carotene in acetonitrile solution was observed to be 15.6 micros. The rate constant for the reaction of triplet energy transfer from triplet 2-acetonaphthone to beta-carotene was calculated to be 1.5 x 10(10) dm3 x mol(-1) x s(-1). Obviously, the triplet beta-carotene has very low excitation energy. Taking the advantage of the photochemical properties of triplet beta-carotene, beta-Carotene has been widely used as energy acceptor to determine the excited state characteristic of other substance. This work extends the understanding of photochemical properties of beta-carotene.

[The study of the pro-nucleating activity of bacteria identified in cholesterol gallstones in model bile systems].

OBJECTIVE: To explore the relationship of bacteria identified in cholesterol gallstones and gallstone formation. METHODS: Observe the bacteria activity in model bile and the influence of bacteria on the cholesterol nucleation time (NT). RESULTS: (1) Model bile were suitable for the growth of E. coli, Pseudomonas aeruginosa, staphylococcus aureus, enterococcus faecalis, clostridium difficile and Clostridium. Propionibacterium acne grew weakly and the growth of Bacteroides fragilis was restrained in model bile. (2) Only pseudomonas aeruginosa and enTerococcus faecalis could ly shorten the cholesterol nucleation time. (3) With pseudomonas aeruginosa or enTerococcus faecalis added in model bile, the formation of cholesterol crystals presented a progressive course of evolution. CONCLUSIONS: Pseudomonas aeruginosa and enterococcus faecalis, not propionibacterium acne, have pro-nucleating ability in model bile.