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Acute abdomen is often a general term for abdominal diseases with acute abdominal pain as the main manifestation. Common clinical acute abdomen includes acute appendicitis, acute cholecystitis, acute cholangitis, acute pancreatitis, gastrointestinal perforation, intestinal obstruction and other diseases, its characteristics are great changes, rapid progress, high misdiagnosis rate, high postoperative complication rate and high mortality rate, accurate diagnosis and early treatment can obtain a good prognosis. With our in-depth understanding of the occurrence and development of acute abdomen diseases and the development of evidence-based medicine, minimally invasive technology plays a pivotal role in the diagnosis and treatment of common acute abdomen. Laparoscopy on diagnosis can clarify disease diagnosis to a large extent. For those who cannot undergo surgery, decompression and drainage under endoscopy provides a diversified plan for treatment decisions. In addition, minimally invasive techniques are also used in etiological treatment and complications. Disease, prevention of recurrence in all aspects, Minimally invasive technology is beneficial to the etiological treatment of biliary pancreatitis, appendicitis and cholangitis, and endoscopic technology is more consistent with the minimally invasive concept in the treatment of complications.
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Objective:To investigate the application value of three-dimensional (3D) printing technology assisted laparoscopic anatomic liver resection of segment 8 (Lap-S8).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 8 liver cancer patients including 7 cases with hepatocellular carcinoma and 1 case with intrahepatic cholangio-carcinoma who underwent 3D printing technology assisted Lap-S8 in the Hunan Provincial People′s Hospital from January 2019 to December 2020 were collected. There were 7 males and 1 female, aged from 49.0 to 80.0 years, with a median age of 56.5 years. Of the 8 patients, 6 cases underwent laparoscopic anatomic liver resection of the entire segment 8, 1 case underwent laparoscopic anatomic liver resection of ventral subsegmental of the segment 8 and 1 case underwent laparoscopic anatomic liver resection of dorsal subsegmental of the segment 8. 3D printing technology was used to assist preoperative evaluation and intraoperative navigation for all 8 patients. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination, internet or telephone interview to detect survival and tumor recurrence of patients after operation up to March 2021. Measurement data with normal distribution were represented as Mean±SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations: all the 8 patients underwent 3D printing technology assisted Lap-S8 successfully, without conversion to open surgery. The operation time, hepatic portal occlusion time and volume of intraoperative blood loss of the 8 patients were (216±41)minutes, (56±11)minutes and 75 mL(range, 50 to 300 mL), respectively. There was no intraoperative blood transfusion in 8 patients, and the surgical margin of the 8 patients was negative. (2) Postoperative situations: the duration of postoperative hospital stay of the 8 patients were (9±3)days. There was no complication such as postoperative hemorrhage, biliary fistula, liver abscess or abdominal infection occurred. (3) Follow-up: all the 8 patients were followed up for 3.0?24.0 months, with a median follow-up time of 12.5 months. During the follow-up, 1 of 8 patients with preoperative diagnosis of recurrent hepatocellular carcinoma developed tumor recurrence at 5 months after operation. The patient underwent laparoscopic surgery followed with the transcatheter arterial chemoembolization and target therapy, and survived with tumor. There was no tumor recurrence in the other 7 patients.Conclusion:3D printing technology assisted Lap-S8 is safe and feasible.
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Background@#Our previous studies have shown that regulatory factor X5 (RFX5), a classical transcription regulator of MHCII genes, was obviously overexpressed in hepatocellular carcinoma (HCC) tumors. However, the role of RFX5 in the carcinogenesis and progress of HCC remains unknown. This study aimed to reveal its biological significance and the underlying mechanism in HCC.@*Methods@#RFX5 mRNA expression level and copy number variation in HCC tumors and cell lines were determined by analyzing deposited data sets in the Cancer Genome Atlas and Gene Expression Omnibus database. The biological significance of RFX5 in HCC was investigated by monitoring the colony formation and subcutaneous tumor growth capacity when RFX5 was silenced with lentiviral short hairpin RNA and CRISPR/Cas9 system in HCC cell lines. The downstream gene transcriptionally activated by RFX5 in HCC cells was determined by chromatin immunoprecipitation and luciferase reporter assay. The involvement of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein theta (YWHAQ) in HCC development was further determined by performing colony formation rescue assay and subcutaneous tumor growth rescue experiment. The association of YWHAQ with recurrence-free survival of patients with HCC was assessed by Kaplan-Meier analysis. Moreover, apoptosis level and the protein level of p53 pathway were determined to reveal the mechanism of RFX5 in driving HCC development.@*Results@#RFX5 was amplified and highly overexpressed in HCC tumor tissues compared with the corresponding non-tumor tissues. The mRNA expression level of RFX5 was significantly correlated with its DNA copy number (r = 0.4, P < 0.001). Functional study demonstrated that RFX5 was required for both clonogenic forming in vitro and subcutaneous tumor growth in vivo of HCC cells. Further study identified YWHAQ, namely 14-3-3 tau, as a key downstream transcriptional target gene of RFX5, which was tightly regulated by RFX5 in HCC. Moreover, overexpression of YWHAQ largely rescued the clonogenic growth of HCC cells that was suppressed by RFX5 knockdown. In addition, overexpression of YWHAQ in primary tumor was linked to poor prognosis of patients with HCC. These results demonstrated that YWHAQ was a downstream effector of RFX5 in HCC. Notably, RFX5-YWHAQ pathway could protect cells from apoptosis by suppressing the p53 and Bax in HCC.@*Conclusion@#RFX5 is a putative HCC driver gene that plays an important role in the development and progression of HCC by transactivating YWHAQ and suppressing apoptosis.
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BACKGROUND@#Our previous studies have shown that regulatory factor X5 (RFX5), a classical transcription regulator of MHCII genes, was obviously overexpressed in hepatocellular carcinoma (HCC) tumors. However, the role of RFX5 in the carcinogenesis and progress of HCC remains unknown. This study aimed to reveal its biological significance and the underlying mechanism in HCC.@*METHODS@#RFX5 mRNA expression level and copy number variation in HCC tumors and cell lines were determined by analyzing deposited data sets in the Cancer Genome Atlas and Gene Expression Omnibus database. The biological significance of RFX5 in HCC was investigated by monitoring the colony formation and subcutaneous tumor growth capacity when RFX5 was silenced with lentiviral short hairpin RNA and CRISPR/Cas9 system in HCC cell lines. The downstream gene transcriptionally activated by RFX5 in HCC cells was determined by chromatin immunoprecipitation and luciferase reporter assay. The involvement of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein theta (YWHAQ) in HCC development was further determined by performing colony formation rescue assay and subcutaneous tumor growth rescue experiment. The association of YWHAQ with recurrence-free survival of patients with HCC was assessed by Kaplan-Meier analysis. Moreover, apoptosis level and the protein level of p53 pathway were determined to reveal the mechanism of RFX5 in driving HCC development.@*RESULTS@#RFX5 was amplified and highly overexpressed in HCC tumor tissues compared with the corresponding non-tumor tissues. The mRNA expression level of RFX5 was significantly correlated with its DNA copy number (r = 0.4, P < 0.001). Functional study demonstrated that RFX5 was required for both clonogenic forming in vitro and subcutaneous tumor growth in vivo of HCC cells. Further study identified YWHAQ, namely 14-3-3 tau, as a key downstream transcriptional target gene of RFX5, which was tightly regulated by RFX5 in HCC. Moreover, overexpression of YWHAQ largely rescued the clonogenic growth of HCC cells that was suppressed by RFX5 knockdown. In addition, overexpression of YWHAQ in primary tumor was linked to poor prognosis of patients with HCC. These results demonstrated that YWHAQ was a downstream effector of RFX5 in HCC. Notably, RFX5-YWHAQ pathway could protect cells from apoptosis by suppressing the p53 and Bax in HCC.@*CONCLUSION@#RFX5 is a putative HCC driver gene that plays an important role in the development and progression of HCC by transactivating YWHAQ and suppressing apoptosis.
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Objective To investigate the expression of MCT8, DCX, SHH and ARC/ARG3. 1 in brain neurons of neonatal rats exposed to thyroid dysfunction in uterus. Methods Wistar pregnant rats were randomly divided into control group and experimental groups that rats were drunk water with 1, 3, or 5 ppm propylthiouracil ( PTU). The thyroid function and morphological changes of PND1 and PND7 were detected. The expression of MCT8, DCX, SHH, ARC/ARG3. 1 protein in cerebral cortex and hippocampus were detected by Western blot or immunohistochemistry. Results (1) The levels of TT4 decreased significantly in PND1 pups of PTU 3 ppm and 5 ppm groups (P＜0.05 or P＜0.01). The TSH levels significantly increased while FT4 levels significantly decreased in pups of PTU 5 ppm group on PND7 ( P＜0. 05). ( 2) The number NV, V, S, and cross-sectional area of thyroid follicles in offspring of PTU groups were significantly higher than those in the control group on postnatal day 1 and 7 (P＜0.05 or P＜0.01, respectively). (3) The expression of MCT8 in cortex and hippocampus gradually increased with the increase dose of PTU on two postnatal days, but there was significant change in PTU 5 ppm group on PND1 ( P＜0.05). The expression of SHH in pup cortex decreased with the increase of PTU exposure dose on PND7. DCX protein expression in the pup cortex on two postnatal days showed an uptrend with the increase of PTU exposure dose. ARC/ARG3.1 protein levels in hippocampal CA1 area of the pups increased significantly in PTU 1 ppm group on PND1 than that in the same-day control group ( P＜0. 05). Conclusion The damaged neurons of neonatal rats exposed to hypothyroidism in utero can be improved with the gradual recovery of thyroid function, but can not be completely restored to normal level.
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Background@#DNA replication and sister chromatid cohesion 1 (DSCC1) (also called DCC1) is a component of an alternative replication factor C complex that loads proliferating cell nuclear antigen onto DNA during S phase of the cell cycle. It is located at 8q24 and frequently amplified in hepatocellular carcinoma (HCC). However, the role of DSCC1 in the carcinogenesis and progress of HCC has not been fully investigated. Here, we aimed to assert the importance of DSCC1 in the HCC.@*Methods@#In this study, copy number variation data and RNA sequencing data were used to calculate the DNA copy number and mRNA expression of DSCC1 in HCC. Quantitative polymerase chain reaction, Western blotting, and immunohistochemistry analysis were used to determine the mRNA and protein level of DSCC1 in HCC. The Kaplan-Meier analysis and univariate and multivariate Cox regression analysis were used to assess the association of DSCC1 with the overall survival (OS) of HCC patients. Moreover, lentiviral shRNA was used to knockdown DSCC1, and then, colony-forming assay, cell cycle assay, and cell proliferation assay were performed to evaluate the impact of DSCC1 silencing on HCC cell lines.@*Results@#We found that DSCC1 was amplified and highly expressed in HCC tumor tissues than in nontumor tissues. We then found that the overexpression of both mRNA and protein of DSCC1 was linked to the bad prognosis of HCC patients. Astonishingly, the protein level of DSCC1 was an independent prognostic factor for OS (hazard ratio, 1.79; 95% confidence interval, 1.17-2.74; P = 0.007). Furthermore, the clonogenic capacity of DSCC1-amplified HCC cell lines (MHCC-97H, MHCC-97L, and Hep3B) was significantly inhibited by transduction of a lentiviral shRNA that targets DSCC1. We also showed that knockdown of DSCC1 induced G0-G1 cell cycle arrest (increased from 60% to more than 80%) and greatly inhibited the proliferation of HCC cell lines.@*Conclusion@#These results suggest that DSCC1 is a putative HCC driver gene that promotes proliferation and is associated with poor prognosis in HCC.
Subject(s)
Female , Humans , Male , Middle Aged , Blotting, Western , Carcinoma, Hepatocellular , Genetics , Pathology , Cell Cycle , Genetics , Physiology , Cell Cycle Checkpoints , Genetics , Physiology , Cell Line, Tumor , Cell Proliferation , Genetics , Physiology , DNA Replication , Genetics , Physiology , Hep G2 Cells , Immunohistochemistry , Liver Neoplasms , Genetics , Pathology , Multivariate Analysis , Proportional Hazards Models , Real-Time Polymerase Chain ReactionABSTRACT
ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.
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Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.
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<p><b>BACKGROUND</b>It has been reported that several baseline polymorphisms of direct-acting antivirals (DAAs) agents resistance-associated variants (RAVs) would affect the treatment outcomes of patients chronically infected with hepatitis C virus (CHC). The aim of this study is to investigate the prevalence of DAAs RAVs in treatment-naÏve GT1b CHC patients.</p><p><b>METHODS</b>Direct sequencing and ultra-deep sequencing of the HCV NS3, NS5A, and NS5B gene were performed in baseline serum samples of treatment-naÏve patients infected with genotype 1b hepatitis C virus (HCVs).</p><p><b>RESULTS</b>One hundred and sixty CHC patients were studied. Complete sequence information was obtained for 145 patients (NS3), 148 patients (NS5A), and 137 patients (NS5B). Treatment-failure associated variants of DAAs were detected: 56.6% (82/145) of the patients presented S122G for simeprevir (NS3 protease inhibitor); 10.1% (14/148) of the patients presented Y93H for daclatasvir and ledipasvir (NS5A protein inhibitors); 94.2% (129/137) of the patients presented C316N for sofosbuvir (NS5B polymerase inhibitor). Nearly, all of the DAAs RAVs detected by ultra-deep sequencing could be detected by direct sequencing.</p><p><b>CONCLUSIONS</b>The majority of genotype 1b CHC patients in China present a virus population carrying HCV DAAs RAVs. Pretreatment sequencing of HCV genome might need to be performed when patients infected with GT1b HCV receiving DAAs-containing regimens in China. Population sequencing would be quite quantified for the work.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Antiviral Agents , Therapeutic Uses , Benzimidazoles , Therapeutic Uses , China , Drug Resistance, Viral , Genetics , Fluorenes , Therapeutic Uses , Genotype , Hepacivirus , Virulence , Hepatitis C , Drug Therapy , High-Throughput Nucleotide Sequencing , Imidazoles , Therapeutic Uses , Polymorphism, Genetic , Genetics , Simeprevir , Therapeutic UsesABSTRACT
<p><b>BACKGROUND</b>Nonalcoholic fatty liver disease (NAFLD) has emerged as the major cause of chronic liver injury. Intestinal barrier plays an important role in the pathogenis of NAFLD. The aim of this article was to assess intestinal immune barrier function during the development of NAFLD.</p><p><b>METHODS</b>Totally 60 male Sprague-Dawley (SD) rats were divided into 2 groups: normal diet (ND) group and high-fat diet (HFD) group. NAFLD rat model was established in the HFD rat group. Portal blood endotoxin level was assessed by limulus test. The percentage of CD4+ cells and CD8+ cells in peripheral blood mononuclear cells (PBMC) and lymphocytes in Peyer's patches (PP) were analysed by flow cytometry. Intestinal secretory immunoglobulin A (SIgA) level was evaluated by enzyme-linked immunosorbent assay. Paired Student's t test was used for the statistic analysis.</p><p><b>RESULTS</b>HFD rats presented with simple steatosis at the 4th and 8th week and progressed to nonalcoholic steatohepatitis at the 12th week. Elevated lipopolysaccharides (LPS) level in HFD rats was observed at the 8th week ((1.54 ± 0.30) times of ND group, P < 0.01). CD4/CD8 ratios in PBMC and PP of HFD rats were increased at the 4th week ((1.50 ± 0.47) and (1.63 ± 0.34) times of ND group, P < 0.05) and decreased at the 8th week ((0.50 ± 0.16) and (0.61 ± 0.26) times of ND group, P < 0.05). At the 12th week, CD4/CD8 ratio ((1.47 ± 0.46) times, P < 0.05) in PP increased to levels observed in the 4th week. Intestinal SIgA expression of HFD rats was remarkably up-regulated at 12th week ((2.70 ± 1.65) times, P < 0.05).</p><p><b>CONCLUSION</b>Liver-gut axis in rats with NAFLD may mediate and improve intestinal immune function by increased CD4/CD8 ratio in PP and increased production of SIgA.</p>
Subject(s)
Animals , Male , Rats , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Diet, High-Fat , Disease Models, Animal , Fatty Liver , Allergy and Immunology , Immunoglobulin A, Secretory , Allergy and Immunology , Intestines , Allergy and Immunology , Non-alcoholic Fatty Liver Disease , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To explore the correlation between single nucleotide polymorphisms (SNPs) of interleukin-28B (IL-28B) gene and the susceptibility to primary hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>A total of 300 histologically confirmed HCC cases (from November 2001 to April 2010) and 310 healthy controls with no history of chronic hepatitis B or hepatocellular carcinoma (2009-2010) were selected from a hospital in Guilin and a hospital in Beijing for this case-control study.139 HCC patients in the case group had complete clinical tracking data. All the subjects were Han Chinese, with no age or gender restrictions.2 ml peripheral blood samples were drawn from each subject with informed consent. SNP of rs12972991, rs4803223, rs8099917 and rs12979860 four loci in IL-28B gene were analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF).</p><p><b>RESULTS</b>The frequencies of C allele at rs12972991, G allele at rs8099917 and G allele at rs4803223 were 6.7% (40/598), 7.9% (47/598) and 10.0% (59/588) respectively in case group; all higher than the corresponding frequencies in control group, separately 2.9% (18/618), 4.1% (25/616) and 3.6% (21/608). The differences were statistically significant (χ2=9.542, 7.858, 20.736, P values all<0.05). The above alleles could increase the risk of HCC, and the OR (95%CI) values were separately 1.67 (1.13-2.46), 1.49 (1.08-2.06) and 2.91 (1.79-4.72). The genotype frequencies of AC+CC at rs12972991, GT+GG at rs8099917, GA+GG at rs4803223 were 13.0% (39/299), 14.7% (44/299) and 19.0% (56/296) respectively in case group; while the frequencies were lower in control group, separately 5.8% (18/309), 8.1% (25/308) and 6.6% (20/304). The differences were statistically significant (χ2=9.319, 6.557, 20.948, P values all<0.05). These genotypes may increase the risk of HCC, and the adjusted OR (95%CI) values were 2.24 (1.31-3.83), 1.81 (1.14-2.88) and 2.90 (1.78-4.70), respectively. The stratified analysis of the clinical data indicated that the frequency of genotype GA+GG at rs4803223 was 50.0% (13/26) in patients of tumor thrombosis in portal vein (TTPV), higher than the frequency of genotype AA (21.1%, 23/109). The difference was statistically significant (χ2=8.965, P=0.003).</p><p><b>CONCLUSION</b>The results suggested that IL-28B gene polymorphisms was correlated to the susceptibility to HCC in Chinese Han ethnic population. Among them, GA + GG genotype at rs4803223 could increase the risk of TTPV in HCC patients.</p>