[Probiotics enhance the efficacy of fecal microbiota transplantation in severe acute liver injury].

Objective: To observe the changes of gut flora in mice, and explore the outcome of fecal microbiota transplantation combined with probiotics in the intervention of severe acute liver injury. Methods: Forty male BALB/c mice were selected and randomly divided into blank control group (10 mice), model group (10 mice), ordinary fecal microbiota transplantation group (10 mice), and fecal microbiota + probiotics transplantation group (10 mice). An intraperitoneal injection of d-galactosamine (D-GalN 3.0g/kg) was given to every group except the blank control group to induce severe acute liver injury model. Simultaneously, ordinary fecal microbiota transplantation group and fecal microbiota + probiotics transplantation group and modeling group were given enema solutions (once a day). After 48 hours, fetched serum was taken to detect alanine transaminase, aspartate transaminase and total bilirubin, and liver tissue was taken for pathological detection. The colonic content was used to extract DNA for 16S V3-V4 high-throughput sequencing. The results of sequencing were analyzed by using bioinformatics analysis; including OTU cluster analysis, α diversity analysis, ß diversity analysis, and linear discriminant analysis effect size (Lefse) to find the bacteria with different colonic content characteristics in different groups of mice. Differences in clinical biochemical indicators between groups were compared using t-test, and the differences between 16S V3-V4 region sequencing results were compared using Wilcoxon test. Results: Model group mice serum biochemical parameters were higher than the other three groups, and the difference was statistically significant (P < 0.05). HE staining of liver sections showed severe inflammatory changes under the microscope in the model group. Ordinary fecal microbiota transplantation group and fecal microbiota + probiotic microbiota transplantation group had low levels of inflammation than the model group. The analysis results of 16S rRNA high-throughput sequencing showed that there was no statistically significant difference in Shannon's index between the blank control and the other three groups. Observed Species difference was statistically significant, and the gut flora composition varied greatly. Species number in the mice gut flora was increased with fecal microbiota transplantation. The results of ß - diversity analysis showed that the difference between the blank control group and the other three groups was greater than that between the disease groups. The difference in the structure of the gut flora of the diseased mice in the fecal microbiota + probiotic transplantation group was mostly butyrate-producing bacteria. Conclusion: Fecal microbiota + probiotics enhance the therapeutic effect of fecal microbiota transplantation, improve liver inflammation, and increase the number of butyrate-producing bacteria in the gut.

Cellular mechanism of Tß4 intervention in liver fibrosis by regulating NF-κB signaling pathway.

OBJECTIVE: To investigate the inhibitory effect of thymosin-ß4 (Tß4) on the activation of the human hepatic stellate cell line (HSC-LX2) induced by interleukin (IL)-1ß. MATERIALS AND METHODS: There were 5 groups in this study, i.e., blank control group, negative control group (SI-NC, empty plasmid), model group (20 ng/ml of IL-1ß), siRNA-Tß4 knockdown group (IL-1ß and si-Tß4) and Tß4 treatment group (IL-1ß and 1000 ng/ml of Tß4). Cell proliferation rate was measured using the Cell Counting Kit-8 (CCK-8) method. The cell cycle change and percentage of apoptotic cells were determined by Propidium Iodide (PI) DNA staining and Annexin V-fluorescein isothiocyanate (FITC) double staining. Cellular nucleic acid levels of p-IKB and nuclear factor-kappa B (NF-κB)/p65 proteins were measured by fluorescent quantitative Real Time-Polymerase Chain Reaction (RT-PCR). Double immunofluorescence staining and Western blot were used to detect nuclear translocation of NF-κB and p65 and levels of cytoplasmic p-IKB protein and nuclear p65 protein. RESULTS: Due to the G0/G1 phase arrest, the number of cells in the Tß4 treatment group increased, compared with the model group and the siRNA-Tß4 knockdown group (p<0.01). In the same between-group comparison, apoptotic rate in the Tß4 treatment group increased significantly (p<0.05). The cellular nucleic acid levels of p-IKB and NF-κB/p65 were markedly higher in the model group and the siRNA-Tß4 knockdown group than in the blank control group (p<0.01). The cellular nucleic acid levels of p-IKB and NF-κB/p65 were remarkably lower in the Tß4 treatment group than in the siRNA-Tß4 knockdown group (p<0.01). The expression levels of NF-κB/p65 and NF-κB/p50 were significantly lower in the Tß4 treatment group. The expression levels of cytoplasmic p-IKB and nuclear NF-κB/p65 were lower in the Tß4 treatment group than in the model group (p<0.01). CONCLUSIONS: Tß4 significantly inhibited IL-1ß-induced HSC-LX2 cell proliferation. The mechanism may involve decreased activation of the NF-κB pathway, decreased expression of p-IKB and nuclear translocation of p65. Therefore, Tß4 had the effect of reversing liver fibrosis.

Ovarian epidermoid cyst: report of eight cases.

We report eight cases of ovarian epidermoid cyst in women 25-57 years of age. All of the cysts had a smooth inner lining composed of stratified squamous epithelium with no evidence of hair or a Rokitansky tubercle. Since these lesions accounted for 0.07% of gynecological specimens in our hospital over a 3-year period, epidermoid cysts of the ovary are not as rare as the literature suggests; some are probably misdiagnosed as dermoid cysts. We believe that ovarian epidermoid cysts represent monodermal and highly differentiated teratomas and should be classified as such.

Krukenberg tumors of the ovary. Ultrastructural, histochemical and immunohistochemical studies of 15 cases.

The ultrastructural, histochemical, and immunohistochemical characteristics of 12 classical signet ring cell Krukenberg tumors (CKT) and three tubular Krukenberg tumors (TKT) were evaluated and related to their possible influence on the ovarian stroma. In CKT, single signet ring cells predominated over lumen-forming cells and contained ultrastructural and histochemical characteristics similar to goblet cells in colonic and ovarian mucinous adenocarcinomas. In TKT, lumen-forming nonsecretory and secretory signet ring cells were prominent. Rare argentaffin cells were found in TKT but not in CKT. Cells in both CKT and TKT produced neutral and sialomucins. The stroma contained extracellular mucin, hypertrophied stromal fibroblasts and myofibroblasts, and in two cases stromal lutein cells with steroidogenic type ultrastructure. It appears that Krukenberg tumors are made up exclusively of intestinal type cells. Based on cell differentiation, TKT is better differentiated than CKT. Hypertrophy and hyperplasia of ovarian stromal cells may occur in response to malignant growth and/or the extracellular mucinous products of malignant cells and may play a role in the control of tumor invasiveness. None of the 15 cases were immunohistochemically positive for chorionic gonadotropin, placental lactogen, or luteinizing hormone. These hormones are suspected to be related to stromal luteinization in KT.