1.Impact of biological function on ovarian clear cell carcinoma ES2 cell line with ARID1A gene expression down-regulating in vitro
Changshuai LYU ; Yinglan ZHANG ; Jinghe LANG
Chinese Journal of Obstetrics and Gynecology 2016;51(3):209-214,215
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.
2.Evaluating gallbladder preservation
Wencai LYU ; Weili FU ; Guiling LANG ; Li JIA
Chinese Journal of Hepatobiliary Surgery 2014;20(3):228-231
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.
3.Prevalence and clinical significance of metabolic disorders of lipids, glucose and uric acid at different fibrosis stages of patients with chronic hepatitis B
Peng WANG ; Zhiqiao ZHANG ; Guotao LYU ; Jing LI ; Lang MING ; Chong ZHENG ; Lewu CHEN
Chinese Journal of Infectious Diseases 2017;35(7):398-402
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.
4.Infection characteristics and drug resistance of methicillin resistant Staphylococcus aureus after liver transplantation
Shaocheng LYU ; Lixin LI ; Qiao WU ; Ren LANG ; Xianliang LI ; Hua FAN ; Xin ZHAO ; Qiang HE
International Journal of Surgery 2017;44(7):456-459
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.
5.Surgical treatment of pancreatic carcinoma with portal vein system invasion
Ren LANG ; Shaocheng LYU ; Qiang HE
Chinese Journal of Hepatobiliary Surgery 2020;26(11):801-804
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.
6.Research progress on the role of endoplasmic reticulum stress in gynaecological diseases
Xinyu HUANG ; Lian RUAN ; Yan WANG ; Chunyu SU ; Mengrong SU ; Wen LANG ; Qizhuang LYU
Immunological Journal 2024;40(4):405-410
Gynecological diseases are one of the important factors that threaten women's reproductive health.Endoplasmic reticulum(ER)stress is a stress response induced by the accumulation of unfolded or misfolded proteins in ER cavity when the female body is in a disease state,and the effects of different stress levels on the female reproductive system and the increase in the risk of female disease will vary greatly.Studies have shown that ER stress is closely related to the occurrence and development of gynecological diseases.Based on the relevant research reports at home and abroad in recent years,this paper summarizes the role of ER stress in several common gynecological diseases to provide new research ideas for preventing,diagnosing,and treating gynecological diseases.
7.A preliminary study of long-term mitochondrial dysfunction in rat brain caused by lipopolysaccharide-induced sepsis.
Juan-Juan LYU ; Zhi-Jiang CHEN ; Dan XIANG ; Gui-Lang ZHENG ; Bin WANG ; Shao-Hua TAO ; Mei-Yan XIE ; Cui LIU ; Jin-Da HUANG ; Qi-Yi ZENG
Chinese Journal of Contemporary Pediatrics 2015;17(8):859-863
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
8.Prognosis and related risk factors in patients with primary liver cancer after liver transplantation
Shaocheng LYU ; Bing PAN ; Lixin LI ; Ren LANG ; Xianliang LI ; Hua FAN ; Ping LI ; Qiang HE
Chinese Journal of Hepatobiliary Surgery 2019;25(7):493-496
Objective To evaluate the prognosis and related risk factors in patients with primary liver cancer after liver transplantation. Methods We retrospectively analyzed the clinical data of 298 patients who underwent liver transplantation between January 2013 and December 2017 at Beijing Chaoyang Hospital of Capital Medical University. 121 patients with primary liver cancer on postoperative pathological diagnosis were included into this study. The patients included 108 males and 13 females, aged from 31 to 70 years, with an average of (52. 7 ± 8. 7) years. The prognosis and the related risk factors on prognosis were analyzed. The survival curve was drawn by the Kaplan-Meier method, and the survival rate was compared by the log-rank test. Multivariate Cox regression was used to analyze the prognostic factors. Results Of the 121 patients who were enrolled in this study, 5 patients died during the perioperative period, making a perioperative mortality rate of 4. 1% (5/121). The remaining 115 patients were followed up and the followed up rate was 95. 0% (115/121). The overall 1-, 2- and 3-year survival rates were 81. 0% , 74. 6% and 70. 2% , respectively. The median survival was 41. 5 months. Multivariate analysis showed that preoperative Child grading (RR=3. 028, 95% CI: 1. 625~5. 643) and microvascular invasion (RR=7. 165, 95% CI: 2. 237~22. 951) were independent risk factors of prognosis. The worse the preoperative Child grading, the worse was the prognosis. The prognosis of patients with microvascular invasion was also poor. Conclusions The overall prognosis of patients with primary liver cancer after liver transplantation was good. Preoperative Child grading and pathological microvascular invasion were the main risk factors of prognosis after liver transplantation carried out for primary liver cancer.
9.Analysis on the application of vascular closure system in portal vein reconstruction of liver transplantation: report of 137 cases
Bing PAN ; Shaocheng LYU ; Lixin LI ; Xin ZHAO ; Zhihua ZHANG ; Ping LI ; Ren LANG ; Qiang HE
Organ Transplantation 2018;9(4):255-260
Objective To analyze the application effect of non-penetrating vascular closure system in portal vein reconstruction of allogenic liver transplantation in adults. Methods Clinical data of 222 patients undergoing allogeneic liver transplantation were retrospectively analyzed. According to whether vascular closure system was used in portal vein reconstruction during operation, all patients were divided into vascular clip group (n=137) and traditional suture group (n=85). Perioperative conditions, clinical prognosis and complications were statistically compared between two groups. Results All patients successfully completed the surgery. The success rate of one-time portal vein anastomosis was 93.4% (128/137) in the vascular clip group. A total of 14 patients died during perioperative period in this study with a mortality rate of 6.3% (14/222). No portal vein-related complications occurred during perioperative period in both groups. The time of portal vein anastomosis in the vascular clip group was (5.6±1.7) min, which was significantly shorter than (10.7±3.6) min in the traditional suture group (P<0.05). The incidence and grade of perioperative complications did not significantly differ between two groups (all P>0.05). Conclusions It is safe and feasible to utilize vascular closure system to reconstruct the portal vein during liver transplantation. Compared with traditional suture, it can effectively shorten the time of portal vein anastomosis.
10.Treatment of acetabular fractures with infra-acetabular screwing
Gang LYU ; Lei MA ; Hui SUN ; Zhiqiang MA ; Bin LANG
Chinese Journal of Orthopaedic Trauma 2021;23(6):507-512
Objective:To investigate the clinical efficacy of infra-acetabular screwing in the treatment of acetabular fractures.Methods:A retrospective analysis was conducted of the 22 patients with acetabular fracture who had been admitted to Department of Trauma and Orthopedics, The Affiliated Hospital of Traditional Chinese Medicine, Xinjiang Medical University from January 2016 to January 2019. They were 16 males and 6 females, aged from 19 to 65 years (mean, 45.2 years). According to Letournel-Judet classification, there were 2 anterior column fractures, 12 anterior plus posterior hemi-transverse fractures, 3 T-shaped fractures and 5 both-column fractures. All patients were treated with infra-acetabular screwing through the ilioinguinal approach. Recorded were the patients' operation time, intraoperative blood loss, reduction quality, fracture union time, hip function and complications.Results:Operation time for this cohort ranged from 115 to 285 min (mean, 160 min), and intraoperative blood loss from 360 to 1,600 mL (mean, 650 mL). By the Matta scoring, fracture reduction was assessed as excellent in 14 cases, as good in 5 cases and as poor in 3 cases, giving an excellent and good rate of 86.4% (19/22). Of this cohort, 21 were followed up from 12 to 45 months (mean, 28.5 months) and one was lost to the follow-up. The fracture healing time for 21 patients ranged from 1.6 to 3.0 months, averaging 2.2 months. No patient had fracture displacement. The Merle d’Aubigné & Postel hip scores at the last follow-up ranged from 8 to18 points (average, 16 points), giving 12 excellent, 6 good, 2 fair, and one poor cases and an excellent and good rate of 85.7% (18/21). Follow-ups observed injury to the lateral femoral cutaneous nerve in one case, deep venous thrombosis of lower limb in 2 cases, superficial wound infection in one case and traumatic arthritis in one case, yielding a total rate of compilations of 23.8% (5/21).Conclusion:Application of infra-acetabular screwing after anatomical reduction of an acetabular fracture can effectively enhance the strength of internal fixation with no risk of fracture re-displacement, conducive to early functional exercise of the patient and leading to good clinical efficacy.