Ferulic Acid Protects Endothelial Cells from Hypoxia-Induced Injury by Regulating MicroRNA-92a.

Materials and Methods: CCK-8 (cell counting kit-8), Western blotting and Annexin V-FITC/PI staining were used to detect cell viability and apoptosis after hypoxia stimulation. The level of microRNA-92 a (miR-92 a) was detected by qRT-PCR.. Then, the assays of flow cytometry and the annexin V/PI staining kit were applied to value the impact of FA on hypoxia-induced cell proliferation, cell cycle distribution, and apoptosis. Furthermore, the inhibitor and mimic of miR-92a were also administrated to explore the role of miR-92a in this process. Student's t-test was used to explore the differences between two groups, while one-way analysis of variance (ANOVA) was used to explore the differences between more than two groups. Results: The results showed that hypoxia stimulation significantly inhibited HUVEC viability and proliferation, such as remarkably decreasing the expression of CDK2, CDK4, and cyclin D1 in HUVECs. The results of annexin V-FITC/PI apoptosis detection showed that hypoxia culture significantly induced HUVEC apoptosis, which indicated that hypoxia stimulation significantly inhibited viability and proliferation of HUVECs but caused cell apoptosis and the expression of miR-92a. Meanwhile, FA remarkably protected HUVECs from hypoxia-induced inhibition of viability and proliferation, as well as the enhancement of apoptosis and miR-92a expression. Furthermore, suppression of miR-92a enhanced the protective effects of FA on hypoxia-induced HUVECs, while activation of miR-92a reversed those effects. Conclusion: Our study reported that FA preserved HUVECs from hypoxia-induced injury via regulating miR-92a, which facilitated the understanding of the protective capacity of FA in hypoxia-caused HUVEC injury.

Supplementation of Bacillus sp. DU-106 Alleviates Antibiotic-Associated Diarrhea in Association with the Regulation of Intestinal Microbiota in Mice.

Bacillus sp. DU-106, a potential probiotic, has been proved to activate innate immunity, reduce hypercholesterolemia, and regulate the gut microbiota of mice. In the present study, we investigated the therapeutic effect of strain DU-106 in antibiotic-associated diarrhea (AAD) via analyzing the changes in gut microbial composition in mice. The results indicated that supplementation of strain DU-106 alleviated gastrointestinal symptoms, improved gut barrier integrity and immunoglobulin-A level of mice with AAD. A 16S rRNA sequencing showed that antibiotics decreased bacterial diversity and the abundances of Alistipes, Roseburia, Hungatella, Eubacterium-xylanophilum, Lachnospiraceae-UCG-001, Intestinimonas, and Lachnospiraceae-NK4A136, but increased the abundance of Klebsiella, Bacteroidota, and Verrucomicrobiota. However, strain DU-106 treatment reversed these alternations in mice with AAD. In conclusion, strain DU-106 could alleviate AAD in association with the regulation of intestinal microbiota and could be used as an alternative treatment for AAD.

Probiotic-fermented black tartary buckwheat alleviates hyperlipidemia and gut microbiota dysbiosis in rats fed with a high-fat diet.

Natural plants fermented with probiotics exert beneficial effects on hyperlipidemia and gut microbiota disorders. This study aimed to investigate the hypolipidemic activity of fermented black tartary buckwheat (FBTB) in rats with hyperlipidemia induced by a high-fat diet (HFD) in association with the regulation of gut microbiota. Probiotic fermentation by Bacillus sp. DU-106 obviously increased the contents of tyrosine, lysine, total flavonoids, total polyphenols, quercetin, and kaempferol in black tartary buckwheat (BTB) and significantly decreased the rutin content. FBTB treatment for 8 weeks significantly decreased the levels of serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol in HFD-induced hyperlipidemic rats. Western blot analysis further confirmed that the protein expression levels of FXR, SREBP1, and PPARα were altered after FBTB treatment. Moreover, FBTB intervention altered the gut microbiota of HFD-fed rats by increasing the relative abundances of Lactobacillus, Faecalibaculum, and Allobaculum and decreasing the relative abundance of Romboutsia. The relative abundance of Allobaculum was positively correlated with the levels of tyrosine, total flavonoids, total polyphenols, quercetin and kaempferol and negatively correlated with that of rutin. These results suggested that FBTB could alleviate hyperlipidemia and gut microbiota dysbiosis in HFD-fed rats.

Retracted: Anticancer Action of Psilostachyin-A in 5-Fluorouracil-Resistant Human Liver Carcinoma are Mediated Through Autophagy Induction, G2/M Phase Cell Cycle Arrest and Inhibiting Extracellular-Signal-Regulated Kinase/Mitogen Activated Protein Kinase (ERK/MAPK) Signaling Pathway.

An editorial decision has been made to retract this manuscript due to breach of publishing guidelines, following the identification of non-original and manipulated figures. Reference: Jun Liu, Yan Liu, Yan Liu, Lei Huang, Guoliang Wang, Jun Wang, Xiangang Xu, Chengxian Shi, Jianzhao Huang: Anticancer Action of Psilostachyin-A in 5-Fluorouracil-Resistant Human Liver Carcinoma are Mediated Through Autophagy Induction, G2/M Phase Cell Cycle Arrest and Inhibiting Extracellular-Signal-Regulated Kinase/Mitogen Activated Protein Kinase (ERK/MAPK) Signaling Pathway. Med Sci Monit 2019; 25:6711-6718. 10.12659/MSM.916635.

Supplementation of Bacillus sp. DU-106 reduces hypercholesterolemia and ameliorates gut dysbiosis in high-fat diet rats.

Gut microbiota modulation by a probiotic is a novel therapy for hypercholesterolemia mitigation. This study initially investigated the potential hypocholesterolemic effect of Bacillus sp. DU-106 in hypercholesterolemic rats and explored its potential relation with gut microbiota. Sprague-Dawley rats received a high-fat diet, or a high-fat diet supplemented with 7.5 × 109 and 1.5 × 1010 CFU/kg bw/day Bacillus sp. DU-106 (low-dose and high-dose groups). At the end of 9 weeks, Bacillus sp. DU-106 treatment significantly decreased the body weight, liver index, and total cholesterol. 16S rRNA sequencing showed that Bacillus sp. DU-106 intervention significantly increased bacterial richness and particularly increased the genus abundance of Turicibacter, Acinetobacter, Brevundimonas, and Bacillus and significantly decreased the abundance of Ralstonia. Metabolomic data further indicated that the supplementation of Bacillus sp. DU-106 remarkably changed the gut metabolic profiles of hypercholesterolemic rats and, in particular, elevated the metabolites of indole-3-acetate, methylsuccinic acid, creatine, glutamic acid, threonine, lysine, ascorbic acid, and pyridoxamine. Spearman's correlation analysis showed the close relation between the different genera and metabolites. In conclusion, Bacillus sp. DU-106 supplement ameliorated high-fat diet-induced hypercholesterolemia and showed potential probiotic benefits for the intestine. KEY POINTS: • A novel potential probiotic Bacillus sp. DU-106 ameliorated hypercholesterolemia in rats. • Bacillus sp. DU-106 supplement regulated gut microbiome structure and richness. • Bacillus sp. DU-106 supplement changed metabolic profiles in high-fat diet rats. • Significant correlations were observed between differential genera and metabolites.

Structural characteristics of a mannoglucan isolated from Chinese yam and its treatment effects against gut microbiota dysbiosis and DSS-induced colitis in mice.

A water-soluble polysaccharide named CYP-1 was isolated from Chinese yam. CYP-1 was characterized as a mannoglucan having a backbone consisting predominately of 1,4-α-linked Glcp branched at O-2, O-3, and O-6 position by t-α-linked Manp with a molecular weight of 2.86 kDa. CYP-1 could inhibit the overproduction of pro-inflammatory cytokines (such as TNF-α and IL-1ß) in LPS-induced RAW 264.7 cells and DSS-induced colitis mice. Oral administration of CYP-1 dramatically alleviated colonic pathological damage, suppressed the activation of colonic inflammatory signaling pathways (such as NF-κB and NLRP3 inflammasome), recovered the mRNA expression of junctional proteins (such as ZO-1, claudin-1, occludin, and connexin-43), and modulated the gut microbiota by decreasing the abundances of Alistipes, Helicobacter, and an unidentified Enterobacteriaceae, in DSS-induced colitis mice. Overall, the present study elucidated that a new polysaccharide structure CYP-1 from Chinese yam and its therapeutic potential as a prebiotic for the prevention of inflammatory bowel disease.

Sacha inchi oil alleviates gut microbiota dysbiosis and improves hepatic lipid dysmetabolism in high-fat diet-fed rats.

Dietary ω-3 polyunsaturated fatty acids (PUFAs) are beneficial for humans against the development of hyperlipidaemia, but the underlying mechanisms are still poorly understood. Here, we demonstrated that oral consumption of sacha inchi oil, which is rich in α-linolenic acid, alleviated dyslipidemia, hepatic steatosis and inflammatory infiltration in high-fat diet (HFD)-fed rats. Sacha inchi oil administration reversed gut microbiota dysbiosis and altered the gut microbiota metabolome and in particular prevented bile acid dysmetabolism caused by a HFD. Sacha inchi oil intake ameliorated hepatic lipid dysmetabolism in HFD-fed rats, via potentiating the biosynthesis and reuptake of bile acids, reducing the de novo lipogenesis, promoting fatty acid beta-oxidation, and alleviating the dysregulation of glycerolipid, glycerophospholipid, and sphingolipid metabolisms. The results showed that dietary sacha inchi oil can alleviate gut microbiota dysbiosis and reduce lipid dysmetabolism in HFD rats, and provide novel insights into the molecular mechanisms by which plant-derived ω-3 PUFAs prevent the development of hyperlipidaemia.

Effect of prophylactic transcatheter arterial chemoembolization on hepatocellular carcinoma with microvascular invasion after R0 resection. A case-control study.

BACKGROUND: Transcatheter arterial chemoembolization (TACE) is thought to prevent recurrence of hepatocellular carcinoma (HCC), but its efficacy is a matter of controversy. OBJECTIVES: We investigated the effect of preventive TACE on the tumor, nodes, metastasis (TNM) classification in cases of stage II HCC (T2N0M0) after R0 resection. DESIGN AND SETTING: Case-control study conducted in a tertiary-level public hospital. METHODS: We analyzed recurrence rates and mortality rates over time for 250 consecutive cases of HCC in TNM classification cases of stage II HCC (T2N0M0) after R0 resection. These cases were divided into patients who underwent TACE (TACE+) and presented microvascular invasion (MVI+; n = 80); TACE+ but did not present MVI (MIV-; n = 100); MVI+ but did not undergo TACE (TACE-, n = 30); and TACE-/MVI- (n = 40). RESULTS: MVI+ patients in the TACE+ group had significantly lower recurrence rates and mortality rates at one, two and three years than those in the TACE- group (all P < 0.05). Among MVI- patients, the TACE+ group did not have significantly lower recurrence rates and mortality rates at one, two and three years than the TACE- group (all P > 0.05). Regardless of whether TACE was performed or not, MVI- patients had significantly lower recurrence rates and mortality rates at two and three years after their procedures than did MVI+ patients (all P < 0.05). CONCLUSION: Recurrence rates and mortality rates for MVI+ patients were significantly higher than for MVI- patients, beyond the first year after TACE. Postoperative adjuvant TACE may be beneficial for HCC patients with MVI.

Effect of prophylactic transcatheter arterial chemoembolization on hepatocellular carcinoma with microvascular invasion after R0 resection. A case-control study

ABSTRACT BACKGROUND: Transcatheter arterial chemoembolization (TACE) is thought to prevent recurrence of hepatocellular carcinoma (HCC), but its efficacy is a matter of controversy. OBJECTIVES: We investigated the effect of preventive TACE on the tumor, nodes, metastasis (TNM) classification in cases of stage II HCC (T2N0M0) after R0 resection. DESIGN AND SETTING: Case-control study conducted in a tertiary-level public hospital. METHODS: We analyzed recurrence rates and mortality rates over time for 250 consecutive cases of HCC in TNM classification cases of stage II HCC (T2N0M0) after R0 resection. These cases were divided into patients who underwent TACE (TACE+) and presented microvascular invasion (MVI+; n = 80); TACE+ but did not present MVI (MIV−; n = 100); MVI+ but did not undergo TACE (TACE−, n = 30); and TACE−/MVI− (n = 40). RESULTS: MVI+ patients in the TACE+ group had significantly lower recurrence rates and mortality rates at one, two and three years than those in the TACE- group (all P < 0.05). Among MVI- patients, the TACE+ group did not have significantly lower recurrence rates and mortality rates at one, two and three years than the TACE- group (all P > 0.05). Regardless of whether TACE was performed or not, MVI− patients had significantly lower recurrence rates and mortality rates at two and three years after their procedures than did MVI+ patients (all P < 0.05). CONCLUSION: Recurrence rates and mortality rates for MVI+ patients were significantly higher than for MVI− patients, beyond the first year after TACE. Postoperative adjuvant TACE may be beneficial for HCC patients with MVI.

SRC-1 and Twist1 are prognostic indicators of liver cancer and are associated with cell viability, invasion, migration and epithelial-mesenchymal transformation of hepatocellular carcinoma cells.

BACKGROUND: Liver cancer is the second leading cause of worldwide cancer-related death, and it has an increasing incidence rate. To investigate the role of SRC-1 and Twist1 in liver cancer and determine their expression in terms of prognosis for patients with liver cancer and in general for hepatocellular carcinoma (HCC) cell lines. METHODS: The present study included a total of 70 patients who underwent liver transplantation or hepatic resection surgeries in our hospital from May 2011 to December 2012. Demographic data and clinical variables as well as alpha-fetoprotein (AFP) and hepatitis B virus (HBV) data were collected. The expression of SRC-1 and Twist1 was determined using immunohistochemistry (IHC). The expression of SRC-1 in different HCC cell lines was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Following SRC-1 silencing by sh-RNA, cell viability, invasion, migration and expression of epithelial-mesenchymal transformation (EMT)-related proteins as well as Twist levels were measured. RESULTS: The expression of SRC-1 and Twist1 was positively correlated in HCC patients. The expression of SRC-1 differed significantly based on patient tumor diameter, tumor-node-metastasis (TNM) grade, and state of liver cirrhosis, and it also differed in patients with dissimilar tumor metastasis conditions, while the expression of Twist1 in patients was significantly correlated with TNM grade and state of liver cirrhosis as well as by the conditions of tumor metastasis. Survival analysis showed that the expression of both SRC-1 and Twist1 were significantly associated with the overall survival (OS) time of HCC patients. Meanwhile, patients with both SRC-1 (+) and Twist1 (+) tissue had the lowest OS, while patients with both SRC-1 (-) and Twist1 (-) tissue had the highest OS. Cox univariate and multivariate analyzes showed that SRC-1 expression, tumor stage and liver cirrhosis were independent risk factors for OS time. SRC-1 was highly expressed in HCC cell lines, and inhibition of SRC-1 had a significantly negative impact on cell viability, invasion, migration and EMT; it also inhibited the expression of Twist. CONCLUSIONS: Expression of both Twist1 and SRC-1 were correlated with clinical outcomes and prognoses for HCC patients, and both Twist1 and SRC-1 were independent risk factors for HCC patient survival conditions. Inhibition of SRC-1 suppressed cell proliferation, invasion, migration and EMT of HCC cells, which might be the result of Twist inhibition.

Anticancer Action of Psilostachyin-A in 5-Fluorouracil-Resistant Human Liver Carcinoma are Mediated Through Autophagy Induction, G2/M Phase Cell Cycle Arrest and Inhibiting Extracellular-Signal-Regulated Kinase/Mitogen Activated Protein Kinase (ERK/MAPK) Signaling Pathway.

BACKGROUND Liver cancer is one of the most common malignancies around the world and one of the major causes of cancer related mortality. The objective of this study was to evaluate the anticancer effect of the natural compound psilostachyin-A on 5-fluorouracil-resistant human liver carcinoma cells and its effects on autophagy, cell cycle, caspase activation, and the ERK/MAPK signaling pathway. MATERIAL AND METHODS Cell Counting Kit 8 (CCK-8) assay was used to evaluate the effects on HepG2 cell viability at different doses of psilostachyin-A. Cell cycle analysis was performed using flow cytometry, and Transwell assay was used to check effects on cell invasion. Transmission electron microscopic studies were done to evaluate autophagy induced by psilostachyin-A, and the western blot method was carried out to evaluate the effects on autophagy and the ERK/MAPK signaling pathway. RESULTS CCK-8 assay revealed that the psilostachyin-A reduced the cell viability of HepG2 cancer cells in a dose dependent manner. Psilostachyin-A also reduced the colony forming potential of HepG2 cells, concentration dependently. The IC50 of psilostachyin was found to be 25 µM. The anticancer effects of psilostachyin-A were due to the induction of autophagy which was accompanied by enhancement of LC3B II expression. Psilostachyin also caused cell cycle arrest by enhancing the accumulation of HepG2 cells in the G2/M phase. Transwell assay showed that psilostachyin-A suppressed the invasion of HepG2 cells. The results also showed that psilostachyin-A could block the ERK/MAPK pathway, indicative of the cytotoxic effects of psilostachyin-A on liver cancer. CONCLUSIONS These preliminary observations suggested that psilostachyin-A might prove beneficial in the treatment of liver cancer.

WITHDRAWN:PPARα Suppresses PD-L1-Mediated Immune Escape by Down-regulating SPP1 in Human Hepatocellular Carcinoma.

Ahead of Print article withdrawn by publisher.

Matrix Metalloproteinase 2 Knockdown Suppresses the Proliferation of HepG2 and Huh7 Cells and Enhances the Cisplatin Effect.

BACKGROUND: This study evaluated the functions of matrix metalloproteinase 2 (MMP2) in hepatocellular carcinoma (HCC) cells and assessed the effects of MMP2 on HCC cell sensitivity to cisplatin. METHODOLOGY: HepG2 and Huh7 cells were cultured. A pre-experiment was performed to explore the optimal transduction conditions of the MMP2-siRNA lentivirus (si-MMP2). Quantitative real-time PCR and western blot assays were performed to measure the expression levels of MMP2 in HepG2 and Huh7 cells. An MTT assay was used to evaluate cell proliferation, and flow cytometry analysis was applied to examine cell apoptosis. A Transwell assay was carried out to assess cell invasion. RESULTS: The optimal virus:cell ratio was 100 multiplicity of infection (MOI) for both cells, and the optimal transduction times for HepG2 and Huh7 cells were 48 h and 72 h, respectively. MMP2 knockdown significantly decreased the mRNA and protein levels of MMP2 in both cell lines (P<0.01). MMP2 knockdown significantly decreased the proliferation and increased the apoptosis of HepG2 and Huh7 cells (P<0.01). Co-treatment with si-MMP2 and cisplatin significantly increased the sensitivity of HepG2 and Huh7 cells to cisplatin (P<0.01). CONCLUSION: MMP2 may act as an oncogene and may be a potential therapeutic target in HCC.

miR-517a promotes Warburg effect in HCC by directly targeting FBP1.

PURPOSE: Hepatocellular carcinoma (HCC) is one of the most aggressive malignancies worldwide. Our aim is to explore the expression and biological function of miR-517a in HCC. MATERIALS AND METHODS: We performed qRT-PCR to detect the expression of miR-517a in clinical samples and cell lines. CKK-8 assay and colony formation assay were employed to detect the miR-517a regulated cell proliferation. Glucose uptake and lactate production were examined to determine the Warburg effect. We also performed ECAR assay using Seahorse system. Luciferase acitivy assay was used to examine the binding of FBP1 3'UTR by miR-517a. RESULTS: miR-517a was upregulated in HCC samples in both genomic and mRNA levels. Moreover, overexpression of miR-517a promoted cell proliferation and Warburg effect. Mechanically, miR-517a could directly target the 3'-UTR of FBP1. In addition, restoring the expression of FBP1 inhibited cell growth. CONCLUSION: We demonstrated that miR-517a acts as an oncogene to promote Warburg effect in HCC, favoring tumor growth, and miR-517a/FBP1 could be a novel target for HCC treatment.

Advantages of laparoscopic common bile duct exploration in common bile duct stones.

OBJECTIVE: To compare the efficacy, safety, and surgical outcomes of laparoscopic common bile duct exploration, endoscopic retrograde cholangiopancreatography, and open common bile duct exploration for treatment of common bile duct stones. METHODS: In total, 210 patients were prospectively randomized into 3 groups: laparoscopic common bile duct exploration, endoscopic retrograde cholangiopancreatography, and open common bile duct exploration. The primary outcome measures were the common bile duct stone clearance rate and the complication rate. The secondary outcome measures were mortality, total costs, and length of hospital stay. RESULTS: The success rates in the laparoscopic common bile duct exploration group (97.14%, 68 out of 70) and open common bile duct exploration group (98.57%, 69/70) were significantly higher than that in the endoscopic retrograde cholangiopancreatography group (85.71%, 60/70, both p < 0.05). The complication rates in the laparoscopic common bile duct exploration group (2.86%, 2/70) and open common bile duct exploration group (1.43%, 1/70) were significantly lower than that in the endoscopic retrograde cholangiopancreatography group (14.29%, 10/70, both p < 0.05). The success rate and complication rate were not significantly different between the laparoscopic common bile duct exploration group and open common bile duct exploration group (both p > 0.05). CONCLUSION: Laparoscopic common bile duct exploration provides an alternative therapeutic approach that was safer and more reliable, allowed for earlier recovery, and provided more cost-effective treatment of common bile duct stones.

Glycosylation change of alpha-1-acid glycoprotein as a serum biomarker for hepatocellular carcinoma and cirrhosis.

AIM: This research is to explore the glycosylation change of alpha-1-acid glycoprotein (AGP) in hepatocellular carcinoma (HCC), cirrhosis and controls. METHODS: The affinity chromatography and lectin affinity techniques were used to separate and enrich glycosylated AGP, and combined with mass spectrometry to identify and relatively quantify the glycopeptides from AGP. RESULTS: The sialylation and fucosylation of AGP were different among HCC, cirrhosis and controls. The highly sialylated and fucosylated peptides from AGP were found in HCC and cirrhosis compared with controls. These glycopeptides showed excellent diagnostic ability to differentiate HCC from cirrhosis (area under the curve >0.9). In addition, these glycopeptides showed significantly different among four HCC stages. CONCLUSION: The sialylation and fucosylation change of AGP may serve as serum biomarker for HCC and cirrhosis.

Real-time Monitoring of Contrast-enhanced Ultrasound for Radio Frequency Ablation.

BACKGROUND: This study compared the realtime monitoring effects of conventional ultrasound and contrast-enhanced ultrasound (CEUS) on evaluating radio frequency ablation (RFA) in a living swine liver model. METHODOLOGY: Liver RFA was performed on 10 young swine. Conventional ultrasound and CEUS were performed immediately. After the animals were sacrificed, ablation lesions were removed to histopathologically examine the range of the lesions. Ablation completeness based on three methods were compared using histopathology as the gold standard. RESULTS: Forty-three ablation lesions were produced in the animals. The horizontal diameter, vertical diameter and ablation lesion area based on conventional ultrasound were all significantly smaller than those based on the gross sample, but no significant differences existed between the results of the CEUS and the gross sample. Histopathology showed that 30 lesions were incompletely ablated and 13 were completely ablated, while CEUS showed that 28 lesions were incompletely ablated and 15 were completely ablated. Compared with histopathology, CEUS had an accuracy of 81.4%, a sensitivity of 83.3%, and a specificity of 76.9%. No significant difference in ablation completeness judgment between CEUS and histopathology was observed. CONCLUSION: CEUS provides a real-time radiological foundation for evaluating RFA lesion ranges and completeness in a living swine liver model.

[Analysis of 216 cases of total laparoscopic hysterectomy].

OBJECTIVE: To investigate the effectiveness and safety of total laparoscopic hysterectomy (TLH). METHODS: A retrospective study of laparoscopic hysterectomy was conducted in this setting. From March 2002 through March 2004, 216 women were subjected to TLH. The average age of the patients was 45.5 years (38-60 years). Out of the 216 patients, 24 had dysfunctional uterine bleeding, 5 atypical endometrial hyperplasia, 139 uterine fibroid, 46 adenomyosis, 2 cervical carcinoma in situ and 36 had a previous lower abdominal surgery. The TLH was carried out using ultrasonic scalpel and the amputated uterus was removed transvaginally. The vagina and peritoneum were closed under laparoscopy. RESULTS: Of the 216 cases who underwent TLH, 23 had bilateral adnexectomy, 36 had ovarian cystectomy, and 54 had adhesiolysis simultaneously. No case was converted to laparotomy. The mean operating time was (103 +/- 35) min. The average amount of blood loss was 83 +/- 45 ml (60-320 ml) during operation. The average hospital stay after operation was (5.3 +/- 1.9) days. There were 4 patients with urinary tract injury in this study population. One bladder perforation was found during operation and repaired under laparoscopy. Two patients had vesicovaginal fistula formation. One ureteral-vaginal fistula was found after operation. The fistula was all closed spontaneously with a prolonged catheter drainage. CONCLUSIONS: TLH appears a safe, effective and reproducible procedure. It is an alternative method for those women who need hysterotomy.

[Influence of p16 gene on the biological behavior of human hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Inactivation of the tumor suppressor gene p16INK4a is one of the most common genetic alterations in human hepatocellular carcinomas (HCC), making it an ideal target gene for treatment of HCC. The objective of this study was to investigate the influence of wild p16 gene on the biological behavior of HCC. METHODS: HCC cell strains SNU-449 (loss of p16 protein expression) and HepG2.2. 15 (positive p16 protein expression) were respectively infected by a retrovirus expression vector of p16 gene (pcLXSN-p16). The stable p16 protein expression cell strains were selected. The biological behaviors of the p16 gene transfected HCC cells were observed. RESULTS: SNU-449 with negative p16 protein expression demonstrated that pcLXSN-p16 treatment significantly inhibited cell growth (the amount of cells at G0-G1 phase increased). However, there was no treatment effect when pcLXSN-p16 was transfected in HepG2.2. 15 which has positive p16 protein expression. Subsequent study in a nude mouse model demonstrated that the p16 gene transfected SNU-449 had a lower succeeding rate of first time establishment of tumors and grew more slowly in the nude mice as compared with non-transfected SNU-449. Moreover, the nude mice inoculated with transfected SNU-449 had a longer survival time than those inoculated with non-transfected SNU-449. CONCLUSION: The transfer of wild p16 gene can inhibit the proliferation and reduce the invasion ability of HCC cells with p16 negative expression, but can not affect the HCC cells with p16 positive expression.