Objective To investigate the efficiency of biological function of AT rich interaction domain 1A (ARID1A) gene silenced by small interfering RNA (siRNA) on ovarian clear cell carcinoma ES2 cell line. Methods (1) The three pairs ARID1A gene siRNA interference fragments siN1 (ARID1A-705), siN2 (ARID1A-1513), siN3 (ARID1A-2282) and one pair negative control were respectively designed, and transfected into ES2 cells by RNA interference max reagent transiently. Reverse transcription (RT)-PCR and western blot methods were used to detect the expression of ARID1A mRNA and protein in ES2 cells transfected with interference fragments respectively. So as to select the best silencing effect of siRNA interference fragment(that was siN3),and then was used in the following experiment. (2) The following experiment were divided into three groups, namely siN3 transfection group, negative control group and blank control group. The proliferative activity of three groups of cells after transient transfection ( 6, 24, 48, 72, 96 hours) was assessed by cell counting kit-8 (CCK-8) assay and expressed as absorbance (A) value; the apoptosis rate of three groups of cells transfected transiently with interference fragment was measured by flow cytometry with annexin V/propidium iodide (PI) staining;the ability of cellular invasion of three groups of cells transfected transiently with interference fragment was tested by transwell experiment;the expression of nuclear factor-kappa B (NF-κB), membrane type-1 matrix metalloproteinase (MT1-MMP) and matrix metalloproteinase-2 (MMP2) protein in ES2 cells transfected transiently with interference fragment was measured by western blot. Results (1) The RT-PCR results showed that the ARID1A mRNA relative expression levels in ES2 cells after transfected transiently with siN1, siN2 and siN3 were 0.007 8±0.005 7, 0.006 8±0.000 3 and 0.002 8±0.000 3 respectively. They were all apparently lower than that in the negative control group (0.034 6 ± 0.001 3;all P<0.01). The western blot results showed that the expression levels of ARID1A protein were 0.439 4±0.000 7, 0.424 4±0.005 0 and 0.386 0±0.005 8 respectively. They were also lower than that in the negative control group (0.732 4 ± 0.030 3; all P<0.01). The siN3 with the highest transfection efficiency was selected to use in the following experiment. (2) The CCK-8 method showed that the proliferative activity of siN3 transfection group cells after transfected transiently at 6 hours was not statistically significant difference compared with those in negative control group and blank control group (0.506 ± 0.010, 0.491 ± 0.006, 0.498 ± 0.009, respectively; all P>0.05). However, the proliferative activity of siN3 transfection group cells after transfected transiently at 24, 48, 72, 96 hours were higher than those in negative control group and blank control group (all P<0.01). The flow cytometry results showed that the apoptosis rate of siN3 transfection group cells was (20.0±3.9)%, which was significantly lower than those in negative control group and blank control group [(31.5 ± 5.0)%, (34.0 ± 4.2)%, respectively;all P<0.05]. The transwell experiment showed that the penetrated cell counts of siN3 transfection group was 60.4±2.9, which was apparently higher than those in negative control group and blank control group (54.2 ± 3.5, 52.1 ± 3.8, respectively; all P<0.01). Western blot experiment showed that the relative expression levels of NF-κB, MT1-MMP and MMP2 protein in siN3 transfection group were respectively 1.85 ± 0.16, 0.37 ± 0.08, 1.38 ± 0.11, which were apparently higher than those in negative control group (0.93±0.11, 0.17±0.05, 0.86±0.06;all P<0.05) and blank control group (0.94 ± 0.04, 0.15 ± 0.08, 0.85 ± 0.10, respectively; all P<0.01). Conclusions It would be to promote the cell doubling time, reduce cell apoptosis and increase the invasive capability in ES2 cells that ARID1A expression was down-regulating by ARID1A mRNA interference. The invasion mechanism may be related to the activation of NF-κB signal transduction pathway, up-regulation of MT1-MMP expression and then promoting the invasion of tumor cells via the up-regulation of MMP2 expression.

A new minimally invasive gallbladder-preserving operation has emerged for the treatment of gallbladder stones,but fundamentally it is a replica of the previous gallbladder preservation method.The gallbladder aids in digestion but is not essential for life.Its function should not be exaggerated and should not instigate trivial reasons for its preservation.Most patients undergoing cholecystectomy maintain a good quality of life.The causes,mechanisms,and prevention of gallstone formation remains mysterious,so a high recurrence rate after gallbladder-preserving operation is difficult to avoid.A full cholecystectomy can avoid recurrence complications and the benefits outweigh the risks.A discussion of the benefits and complications must be initiated to evaluate both treatment modalities for gallstones.

Objective To explore the prevalence and clinical significance of metabolic disorder of lipids, glucose and uric acid at different fibrosis stages of patients with chronic hepatitis B (CHB).Methods From January 2006 to December 2014, 1 812 CHB patients in Department of Infectious Diseases, Shunde Hospital of Southern Medical University were retrospectively enrolled and analyzed.All biochemistry indexes were obtained by automatic biochemical instrument.Polymerase chain reaction was used for detecting serum hepatitis B virus (HBV) DNA, and particles immune detection kit was used for detecting hepatitis B e antigen (HBeAg).In statistical analyses, chi-square test, nonparametric test and Logistic analysis were used.Results The metabolic disorder prevalence in 1 812 CHB patients was as follows, 455 cases (25.1%) with decreased high density lipoprotein, 435 cases (24.0%) with increased uric acid, 342 cases (18.9%) with increased total cholesterol, 254 cases (14.0%) with increased triglyceride, 171 cases (9.4%) with decreased apolipoprotein A, 165 cases (9.1%) with increased apolipoprotein B, 162 cases (8.9%) with increased low density lipoprotein and 117 cases (6.5%) increased fasting blood glucose.Patients who had mild liver fibrosis tended to have metabolic disorders of uric acid (26.4%), total cholesterol (22.8%) and high density lipoprotein cholesterol (20.5%).Patients who had moderate liver fibrosis tended to have metabolic disorders of high density lipoprotein (27.2%) and uric acid (20.9%).Patients who had severe liver fibrosis tended to have metabolic disorders of high density lipoprotein (33.6%) and uric acid (22.2%).Multivariate Logistic regression analysis showed that inflammation activty (OR=17.31, 95% CI: 13.410-22.336, P=0.001), age (OR=1.019, 95%CI:1.005-1.035, P=0.010), sex (OR=1.497, 95% CI: 1.061-2.111, P=0.022), apolipoprotein A (OR=0.50, 95% CI: 0.281-0.892, P=0.019) and HBV DNA (OR=0.904, 95% CI: 0.858-0.952, P=0.001) may be independent predictors of moderate and severe liver fibrosis.Conclusions CHB patients with mild liver fibrosis tend to have metabolic disorders of uric acid, total cholesterol and high density lipoprotein cholesterol;patients with moderate liver fibrosis tend to have metabolic disorders of high density lipoprotein and uric acid;and patients with severe liver fibrosis tend to have metabolic disorders of high density lipoprotein and uric acid.

Objective To investigate the infection characteristics and drug resistance of methicillin resistant Staphylococcus aureus after liver transplantation.Methods Retrospectively analyzed the clinical data of 170 patients who underwent liver transplantation in Beijing Chaoyang Hospital,Capital Medical University between January 2011 and April 2016.The incidence,pathogenic characteristics,distribution and drug resistance of methicillin resistant Staphylococcus aureus were analyzed.Results In this research,the methicillin resistant Staphylococcus aureus infection occurred in 23 cases after liver transplantation,and the incidence rate was 13.53％ (23/170).There were 27 strains of pathogens had been isolated,and 74.07％ (20/27) of pathogens were isolated from peritoneal drainage fluid.The most common methicillin resistant Staphylococcus aureus were Staphylococcus haemolyticus,Staphylococcus epidermidis and Staphylococcus hominis,accounting for 29.63％ (8/27),22.22％ (6/27) and 14.81％ (4/27).Drug sensitivity results indicated that the methicillin resistant Staphylococcus aureus were completely resistant to Penicillin,Oxacillin and Methicillin antibiotics.And the pathogens were extensively drug-resistant to Ciprofloxacin,Levofloxacin and Moxifloxacin,the resistance rates were 63％ (15/24),63％ (15/24) and 58％ (14/24).But the pathogens were sensitive to Teicoplanin,Vancomycin and Linezolid,and there have not been detected drug-resistant bacteria.Conclusions The infection of methicillin resistant Staphylococcus aureus is one of the most common pathogens after liver transplantation.Routine prophylactic antibiotics are not useful for the treatment,however,Teicoplanin,Vancomycin and Linezolid can be used as the first choice of treatment.

Pancreatic carcinoma is one of the most common digestive malignant tumor. It has a high degree of malignancy and poor prognosis. The overall 5-year survival rate is less than 10%. Portal vein system invasion is one of risk factors for poor long-term prognosis of pancreatic carcinoma, and radical surgery is the only effective way to achieve long-term survival. According to the clinical practice, our center proposed a new classification of pancreatic carcinoma with portal vein system invasion for the purpose of reconstructing the portal vein system. We also formulated treatment strategies for different classification and carried out operation of allogeneic vascular replacement in clinical practice. This paper introduces our relevant clinical experience.

OBJECTIVETo preliminarily investigate the long-term structural and functional injuries of mitochondria in rat brain caused by sepsis.

METHODSWistar rats were randomly assigned into sepsis and control groups. A rat model of sepsis was prepared by an intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) of gram-negative bacteria, and the survival assay was performed. Eight rats in the sepsis group were sacrificed at 12, 24, 48, or 72 hours after LPS injection, while rats in the control group were sacrificed after an intraperitoneal injection of an equal volume of normal saline. Mitochondria were extracted from rat brain tissue. Mitochondrial membrane potential (MMP) and mitochondrial swelling level were determined by flow cytometry, and the activities of electron transport chain complexes (I-V) were measured using enzyme assay kits. Hematoxylin-eosin (HE) staining and electron microscopy were used to observe morphological changes in brain tissue and mitochondria.

RESULTSThe sepsis group had a significantly lower survival rate than the control group (P<0.01). The MMP and activities of electron transport chain complexes (I-V) in the sepsis group, which were significantly lower than those in the control group (P<0.05), were reduced to the lowest levels at 48 hours and partially recovered at 72 hours. The mitochondrial swelling level in the sepsis group, which was significantly higher than that in the control group (P<0.05), increased to the peak level at 48 hours and partially recovered at 72 hours. Hematoxylin and Eosin staining revealed substantial damages in the structure of brain tissue, and electron microscopy showed mitochondrial swelling, and vacuolization in a few mitochondria.

CONCLUSIONSIn the rat model of LPS-induced sepsis, both structural and functional injuries are found in cerebral mitochondria, and achieve the peak levels probably at around 48 hours.

Animals ; Brain ; pathology ; physiopathology ; ultrastructure ; Lipopolysaccharides ; toxicity ; Male ; Membrane Potential, Mitochondrial ; Mitochondria ; physiology ; ultrastructure ; Rats ; Rats, Wistar ; Sepsis ; chemically induced ; mortality ; physiopathology

OBJECTIVE To investigate the effect of amifostine(Amf)on the differentiation of human megakaryocyte cell line-Dami. METHODS Dami cells were treated with Amf 0.01-5.0 mmol · L-1 for 12 d. Dami cells were counted every day for the growth curve:only cells with a diameter>20μm. The platelet demarcation membrane system was observed by transmission electron microscopy. The expression of CD33,CD34,CD41a and DNA ploidy was detected by flow cytometry. RESULTS Amf 0.1-1.0 mmol · L-1 promoted the differentiation of Dami cells ,but inhibited their proliferation at a concentration>1.0 mmol · L-1. When these cells were treated with Amf 1.0 mmol · L-1 for 12 d,the platelet demarcation membrane system was observed,the percentage of cells with a diameter >20 μm was increased by 24.6%(P<0.01),the expression of CD41a was increased by 11.9%,while the expression of CD33 was decreased by 13.6%(P<0.05). Polyploidy cells(16N)were observed,and 4N,8N and 16N cells were increased to 31.56%,8.83% and 3.43%,respectively(P<0.05). CONCLUSION Amf 0.1-1.0 mmol · L-1 can promote the differentiation of Dami cells,but inhibit their proliferation at a high concentration(>1.0 mmol·L-1).

Objective To investigate the clinical characteristics and the related risk factors of pulmonary infection in patients after liver transplantation.Methods The clinical data of 298 patients who underwent liver transplantation in the Beijing Chaoyang Hospital between January 2013 and December 2017 were retrospectively stndied.The patients were divided into the infection group (n =58) and the control group (n =240) according to whether they had pulmonary infection.The characteristics,diagnosis,treatment,prognosis and risk factors of pulmonary infection were analyzed.Results Pulmonary infection occurred in 58 patients (19.5％) after liver transplantation.Eight patients died in the infection group of pulmonary infection.The mortality rate was 13.8％.The most common pathogenic bacteria were Acinetobacter bauman,Pseudomonas aeruginosa and Klebsiella pneumoniae,which accounted for 28.8％,20％ and 13.8％,respectively for the pulmonary infection.Multivariate analysis showed that the neutrophil ratio,and intensive care unit (ICU) hospitalization time were independent risk factors of pulmonary infection (P ＜ 0.05).Conclusions Pulmonary infection was common after liver transplantation.The treatment strategy should be the use of rational antibiotics and intensive pulmonary management.The neutrophil ratio and ICU hospitalization time were the risk factors of pulmonary infection.

Objective To analyze the application and effect of non-penetrating vascular and tubular tissue closure system in radical resection of adenocarcinoma of pancreatic head combined with venous resection and reconstruction.Methods A retrospective analysis was made on the clinical data of 78 patients with pancreatic head cancer who underwent radical resection of venous vessels from Feb 2014 to Feb 2018.According to the intraoperative venous resection and anastomosis,the patients were divided into vascular clip group (41 cases) and traditional suture group (37 cases).The preoperative data,intraoperative and postoperative recovery of the two groups were analyzed and compared.Results There were no perioperative deaths and no significant differences in perioperative complications between the two groups (P ＞ 0.05).The anastomotic time of the clip group was (18.6 ± 3.3) min,which was significantly shorter than that of the traditional suture group (39.7 ± 8.5) min,(P ＜ 0.05).There were no significant differences in operation time,intraoperative bleeding volume,ICU time,hospital stay and anastomotic stenosis between the two groups(P＞ 0.05).Conclusion It is safe and feasible to use vascular clip in venous vascular reconstruction in radical resection of carcinoma of the head of the pancreas combined with venous vascular resection.

OBJECTIVE: To explore the genetic basis for a child suspected for hypokalemic periodic paralysis. METHODS: Clinical data of the patient was collected, and venous blood samples were taken from the patient and his parents for the extraction of genomic DNA. Next generation sequencing (NGS) with target capture was carried out to detect potential variants. Suspected variants were validated by Sanger sequencing. RESULTS: The child developed fatigue without obvious reason at the age of 15. Laboratory test revealed hypokalemia but normal serum magnesium. Genetic testing discovered that he has carried two variants in the SLC12A3 gene, namely c.179C>T and c.539C>A. The patient was diagnosed with Gitelman syndrome. CONCLUSION For children with hypokalemia, genetic testing should be considered for the differential diagnosis of Gitelman syndrome from hypokalemia due to other causes.