ObjectiveTo study the effect and mechanism of lipopolysaccharide (LPS) on osteoclasts formation and its bone resorption function.MethodsBone marrow-derived macrophages (BMMs) were extracted from the marrow of femur and tibia of 4-week-old male C57BL/6 mice. Flow cytometry was used to detect BMMs. The effect of different concentrations of LPS (0, 100, 200, 500, 1 000, 2 000 ng/mL) on BMMs activity was examined by cell counting kit 8 (CCK-8) activity test. In order to investigate the effect of LPS on osteoclastogenesis, BMMs were divided into macrophage colony-stimulating factor (M-CSF) group, M-CSF+receptor activator of nuclear factor κB ligand (RANKL) group, M-CSF+RANKL+50 ng/mL LPS group, M-CSF+RANKL+100 ng/mL LPS group. After the completion of culture, tartrate resistant acid phosphatase (TRAP) staining was used to observe the formation of osteoclasts. In order to investigate the effect of LPS on the expression of Connexin43, BMMs were divided into the control group (M-CSF+RANKL) and the LPS group (M-CSF+RANKL+100 ng/mL LPS); and the control group (M-CSF+RANKL), 50 ng/mL LPS group (M-CSF+RANKL+50 ng/mL LPS), and 100 ng/mL LPS group (M-CSF+RANKL+100 ng/mL LPS). The expressions of Connexin43 mRNA and protein were detected by Western blot and real-time fluorescent quantitative PCR, respectively. In order to investigate the effect of LPS on osteoclast bone resorption, BMMs were divided into M-CSF group, M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group. Bone absorption test was used to detect the ratio of bone resorption area.ResultsThe flow cytometry test confirmed that the cultured cells were BMMs, and CCK-8 activity test proved that the 100 ng/mL LPS could promote the proliferation of BMMs, showing significant differences when compared with the 0, 200, 500, 1 000, and 2 000 ng/mL LPS (P<0.05). TRAP staining showed no osteoclast formation in M-CSF group. Compared with M-CSF+RANKL group, the osteoclasts in M-CSF+RANKL+50 ng/mL LPS group and M-CSF+RANKL+100 ng/mL LPS group were larger with more nuclei, while the osteoclasts in M-CSF+RANKL+100 ng/mL LPS group were more obvious, and the differences in the ratio of osteoclast area between groups were statistically significant (P<0.05). Western blot result showed that the relative expression of Connexin43 protein in LPS group was significantly higher than that in control group (P<0.05). Real-time fluorescent quantitative PCR showed that the relative expression of Connexin43 mRNA in control group, 50 ng/mL LPS group, and 100 ng/mL LPS group increased gradually, and the differences between groups were statistically significant (P<0.05). Bone resorption test showed that osteoclast bone resorption did not form in M-CSF group, but the ratio of bone resorption area increased gradually in M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group, and the differences between groups were statistically significant (P<0.05).ConclusionLPS at concentration of 100 ng/mL can promote the expression of Connexin43, resulting in increased osteoclastogenesis and enhanced osteoclastic bone resorption.
Developmental and epileptic encephalopathy (DEE) is a group of diseases that severely affects the neurological development of children, characterized by frequent seizures and significant neurodevelopmental impairments. These diseases not only impact the quality of life of affected children but also impose a heavy burden on families and society. In recent years, the development of brain network theory has provided a new perspective on understanding the pathological mechanisms of DEE, especially the role of structural and functional brain networks in the process of epilepsy. This review systematically summarized the research progress of structural and functional brain networks in DEE, highlighted their importance in seizure activity, disease progression, and prognosis evaluation.
Objective To study the effect on expression of high mobility group box-1 （HMGB1） mRNA for the expression of zonula occludens-1 （ZO-1） in ileum tissues， and to explore the possible mechanism of intestinal mucosal barrier injury in rats with acute necrotizing pancreatitis （ANP）. Methods Ninety-six male Wistar rats were divided randomly （random number method） into ANP group， ethyl pyruvate （EP）group， and sham operation group. Eight rats of 3 groups were killed to get abdominal aortic blood and ileal tissues at 6， 12， 24， and 48h after operation， respectively.The levels of plasma amylase （AMY） ， D-lactate acid， and the activity of malonyl dialdehyde （MDA） in the ileum tissues were determined by using automatic biochemical analyzer， improved enzymatic spectrophotometry， and thiobarbituric acid （TAB） colorimetry respectively. The pathological changes of ileum tissues were observed under microscopy by HE staining， the expression of ZO-1 protein in ileum tissues was observed by immunohistochemistry （SP method）， and the expressions of HMGB1 mRNA and ZO-1 mRNA in ileum tissues were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results Compared with ANP group at the same time， levels of AMY， D-lactate acid， and MDA in ileum tissues of EP group were all significantly lower （P＜0.05）. The expression level of HMGB1 mRNA increased at 6 h while ZO-1 mRNA decreased in ANP group. Compared with ANP group at the same time， the expression level of HMGB1 mRNA of EP group was significantly lower while ZO-1 mRNA was higher （P＜0.05）， and the pathological damage in ileum tissues was lighter. Conclusions The decreased expression of ZO-1 in ileum tissues is one of the vitalcauses for intestinal mucosal barrier injury in ANP， and it probably occurs in case of the excessive expression of HMGB1.
Objective To investigate the clinical effect of retroperitoneal laparoscopic dismembered pyeloplasty on ureteropelvic junction obstruction (UPJO). Methods From November 2010 to September 2015, a total of 28 cases (21 males and 7 females) with ureteropelvic junction obstruction (UPJO) underwent retroperitoneal laparoscopic dismembered pyeloplasty. All the 28 patients had a previous history of different degrees of lumbar pain, which was confirmed as typical UPJO by imaging examination. Operation method: following complete renal pelvis and upper ureter isolation, the renal pelvis was clipped into a trumpet-shaped mouth, 1/3 of the upper renal pelvis was temporarily kept without section. In the inferior margin of renal pelvis, longitudinal cut procedure was operated in the ureter and across the stenotic segment in about 1–2 cm; and then, anastomosis of the ureter and the renal pelvis at the lowest was performed, to maintain the continuity of the renal pelvis and ureter. Subsequently, the stenosed segment of ureter and the dilatant renal pelvis were removed, the posterior ureteropelvic anastomosis was made with an interrupted suture, and antegrade stenting of double J tube was further performed, followed by a continuous suture of the anterior wall. Results The procedure was successfully accomplished in all patients whithout conversion to open surgery. The operating time was 90-240 minutes with the median time of 160 minutes. The estimated blood loss was 20-70 mL with the median of 50 mL. No severe surgical complication occurred in perioperative period. The 3-24 -month follow up showed that symptoms of lumbar pain were relieved or disappeared in all the incorporated patients. Postoperative radiographic examination showed no stomal stenosis at the ureteropelvic junction, and the hydronephrosis was alleviated. Conclusions Retroperitoneal laparoscopic dismembered pyeloplasty for UPJO is an effective, safe, and minimally invasive surgical technique. The key to the success of the operation is to the accurate renal pelvis clipping, tension- and torsion-free anastomosis of the lowest position of the renal pelvis and the ureter, and the reconstruction of a new funnel-shaped ureteropelvic junction.
ObjectiveTo analyze the outcomes of patients with one-stage Fontan procedure and short-term prognosis at a single institute. Method We retrospectively analyzed clinical records of 116 patients with one-stage Fontan procedure in our hospital from January 2008 through September 2013. There were 77 males and 39 females (36%), with median age 6.27 years (ranged 3.15 to 17.47 years) at the time of surgery and the median weight at 17.5 kg (ranged 10.0 to 80.0 kg). There were 55 patients with standard extracardiac conduit, 13 patients with direct extracardiac connection, 17 patients with intra-atrial or intra/extracardiac conduit, and 31 patients with lateral tunnel. ResultsMedian cardiopulmonary bypass time was 124 (61-256) minutes. Median cross-clamp time was 60 (19-152) minutes. There were six early deaths (5.1%). The overall median time of the cardiac intensive care unit stay was 4 (1-17) days, with a median ventilator support of 7.3 (1.0-181.0) hours. The mean room air saturation was 90.00%±4.68% before discharge. Median length of chest tube drainage was 10 (4-45) days. Multiple logistic analysis confirmed that heterotaxy syndrome was the only independent predictor for postoperative renal insufficiency. Operations with aortic cross-clamping (OR=26.184, 95% CI 1.712-400.451), preoperative sinus mode dysfunction (OR=6.777, 95% CI 1.495-30.721) and cross-clamp time over 60 minutes (OR=1.036, 95% CI 1.002-1.076) were predictors for prolonged chest tube drainage. A total of 110 patients were followed up for 17 (8-47) months with 2 deaths and 1 with thrombosis. No reoperation occurred. ConclusionThe one-stage Fontan procedure can be performed with satisfactory outcomes. Staged strategies for operations may be appropriately loosen for selected elder children.
Objective To assess clinical results of three different conduit materials （Gore-Tex synthetic graft，bovinejugular vein and autologous pericardium）for palliative right ventricle-to-pulmonary artery （RV-PA） shunt，and explore the correlation between suitable conduit size and patients’ body weight and McGoon ratio. Methods We retrospectively analyzed clinical data of 24 patients with congenital heart diseases who underwent palliative RV-PA shunt in Department of Pediatric Cardiovascular Surgery of Fu Wai Cardiovascular Hospital from July 2010 to July 2012. There were 11 males and 13 females with their age ranging from 60 days to 6 years and body weight of 10.22±7.41 kg. There were 22 patients with pulmonary atresia and ventricular septal defect （PAVSD），1 patient with tetralogy of Fallot （TOF） and 1 patient with doubleoutlet right ventricle （DORV）. Among different conduit materials，autologous pericardium was used for 17 patients，Gore-Texsynthetic graft was used for 5 patients，and bovine jugular vein was used for 2 patients. Conduit size and children’s body weight were analyzed with linear regression，then the equation was corrected with McGoon ratio. Results There was no perioperative death. Postoperative percutaneous saturation （SpO2）of the 24 children was 20.37%±28.33% higher than preoperative SpO2 . Electrocardiogram showed sinus rhythm in all the patients. Twenty-three patients were NYHA classⅡ，and 1 patient was NYHA classⅢ. Postoperative mechanical ventilation time of patients with autologous pericardium were significantly shorter than those of patients with other 2 materials （P=0.017）. Sixteen patients were followed up from 10 months to 2 years after discharge，including 12 patients with autologous pericardium，3 patients with Gore-Tex synthetic graft and 1 patient with bovine jugular vein. During follow-up，McGoon ratio of patients with autologous pericardium，Gore-Tex synthetic graft and bovine jugular vein were 1.98±0.46，1.83±0.33 and 1.68 respectively，all of which weresignificantly higher than preoperative McGoon ratio （P＜0.05）. Six patients underwent radical corrective surgery，including5 patients with autologous pericardium and 1 patient with Gore-Tex synthetic graft. There was no complication directly related to surgery during follow-up. Linear regression was performed to form an equation between suitable conduit size and patients’ body weight：conduit diameter （mm）=0.327×body weight （kg）＋4.599. McGoon ratio，conduit size and equationresult were compared to find a practical choice of conduit size. If McGoon ratio＜0.8，the first integer greater than the equation result could be chosen. If McGoon ratio＞1.2，the first integer less than the equation result could be chosen. If 1.2＞McGoon ratio＞0.8，the first integer either less or greater than the equation result could be chosen. Group analysis showed that patients who recovered better postoperatively were those whose conduit sizes were closer to equation results as well as equation results corrected with McGoon ratio. Conclusion All the 3 materials can be conventionally chosen for RV-PA shunt. Appropriate conduit size can be decided upon patients’ body weight and McGoon ratio for RV-PA shunt.
Objective To observe the influence of connexin 43 (Cx43) on bystander effects induced by cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-tk) coexpression suicide genes system in human cholangiocarcinoma QBC939 cells and transplantation tumors in nude mice. Methods In vitro, the CD+tk+ and CD+tk+Cx+ cells were respectively treated with 5-fluorocytosine (5-FC) and ganciclovir (GCV). The cytotoxic efficacy was evaluated by microculture tetrajolium test (MTT) method. In order to investigate the influence of Cx43 on bystander effects, the volumes of transplantation tumors of the CD+tk+ and CD+tk+Cx+ cells were measured before and after application of 5-FC and GCV. Results CD and tk gene were stably expressed in transfected QBC939 cells. Increasing expression of Cx43 was determined by testing for the presence of Cx43 mRNA by RT-PCR and the presence of Cx43 protein by Western blot in CD+tk+Cx+ cells. The killing effect of 5-FC and GCV on CD+tk+Cx+ cells was more effective than that on CD+tk+ cells both in vitro and in vivo. Conclusion Double suicide genes system CD/5-FC+tk/GCV could induce remarkable killing effect on cholangiocarcinoma cells in vitro and transplantation tumors in vivo. The cotransfection of Cx43 gene is able to enhance the bystander effects and the inhibition of carcinoma cells.
ObjectiveTo isolate and culture cartilage derived stem cells from different subtypes of cartilages, and to identify their characteristics. MethodsCartilage derived stem cells were isolated from different subtypes of cartilages (auricle cartilage, articular cartilage, and intervertebral cartilage) by using adhesive method of fibronectin. The expressions of positive surface markers (CD29 and CD90) and negative surface markers (CD34 and CD45) in cartilage derived stem cells were detected via flow cytometry. The single cell colony-forming efficiency of cartilage derived stem cells was determined by clonal formation unit test; the multipotent differentiation capacity was identified by chondrogensis, osteogenesis, and adipogenesis induction. RT-PCR was used to test the expression of osteogenic, chondrogenic, and adipogenic genes; and bone marrow mesenchymal stem cells (BMSCs) served as control. ResultsThree cell populations were successfully isolated from different subtypes of cartilages, which could express CD29 and CD 90 highly, but did not express CD34 and CD45. After 2 weeks of culture, single cartilage derived stem cell could form single cell colony. In addition, cartilage derived stem cells had high chondrogenesis, osteogenesis, and adipogenesis potentials. After osteogenic induction, the expressions of collagen type Ⅰ and collagen type X in articular and intervertebral cartilage stem cells were significantly higher than those in BMSCs (P<0.05), while there was no significant difference between auricular cartilage stem cells and BMSCs (P>0.05). The expressions of Aggrecan and collagen type Ⅱ in cartilage derived stem cells after chondrogenic induction were significantly higher than those in BMSCs (P<0.05). While the ability of adipogenic differentiation was lower than that in BMSCs, but no significant difference was found (P>0.05). ConclusionCartilage derived stem cells in different subtypes of cartilages possess typical characteristics of stem cells.
ObjectiveTo summerize the early results of total cavopulmonary connection (TCPC) procedure with an extracardiac conduit in adults with congenital heart disease, and assess risk factors for postoperative delayed recovery in ICU. MethodsWe retrospectively analyzed the clinical data of 20 adult patients underwent TCPC operation with extracardiac conduit in Fu Wai Hospital between January 2012 and December 2014. There were 14 female and 6 male patients at age of 16 to 33 (20.45±4.33) years. ResultsThere was no hospital mortality. The time of ICU stay was 4.4±1.7 days. And time of hospital stay was 32.5±21.6 days. Morbidities included prolonged pleural effusion lasting more than 7 days in 12 patients (60.0%), new arrhythmias in 3 patients (15.0%), reexploration for bleeding in 3 patients (15.0%), surgical wound poor healing in 1 patient (5.0%). Dopamine and calcium were used in all the patients in the ICU, epinephrine in 18 patients, milrinone in 11 patients. Risk factors for postoperative delayed recovery in ICU were preoperative arrhythmias (P=0.02), cardiopulmonary bypass time longer than 120 min (P=0.04), plasma applications more than 2 000 ml (P=0.01), absence of fenestration (P=0.04), and pleural effusion lasting longer than 7 days (P=0.04). ConclusionThe TCPC procedure with an extracardiac conduit can be performed in adults with encouraged early results. Actively vasoactive drugs to maintain circulation early in ICU has good results for the patient's recovery.