BACKGROUND:It has been proved that erythropoietin can promotes angiogenesis in injured tissue, which is closely related to the proliferation and differentiation of endothelial progenitor cel s. However, the involved mechanism remains unclear yet.
OBJECTIVE:To investigate the effect of erythropoietin on the function and activity of bone marrow-derived endothelial progenitor cel s in mice, and to explore the signal pathway.
METHODS:The endothelial progenitor cel s from the bone marrow of mice were separated by means of density gradient centrifugation and then cultured. The cel s were preconditioned by specific inhibitor of PI3K
(LY294002), and were divided into the fol owing groups:EGM-2 group, three erythropoietin preconditioned groups (the concentrations of erythropoietin in medium were 1, 5, 10 U/mL respectively), erythropoietin+LY group (10 U/mL erythropoietin and 10 mmol/L LY294002 in medium), LY group (10 mmol/L LY294002 in medium), dimethyl sulfoxide group (1 mL/L dimethyl sulfoxide in medium). The cel proliferation and apoptosis were evaluated by cel counting kit-8 and flow cytometry respectively. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cel lysates were detected by the method of ELISA, and the expressions of Akt and p-Akt were by western blot assay.
RESULTS AND CONCLUSION:Erythropoietin could promote the proliferation of endothelial progenitor cel s in a dose-dependent manner, which was, however, completely inhibited by LY294002. The apoptosis rate in the erythropoietin preconditioned groups was significantly lower than that in the erythropoietin+LY group. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cel lysates of LY group and erythropoietin+LY group were significantly lower than those in the erythropoietin groups. There was no difference in Akt expression found in each group, while the p-Akt expression in the erythropoietin+LY group was significantly lower than that in the erythropoietin groups. The above results reveal that erythropoietin can promote the proliferation of endothelial progenitor cel s and decrease the cel apoptosis, which is depending on PI3K/Akt signal pathway.

Objective To summary the experience of T4 esophageal carcinoma surgery and to explore the methods and operating skills on descending thoracic aortic resection with prosthetic vascular graft replacement in patients with T4 locally advanced esophageal carcinoma invading descending thoracic aorta.Methods From Jan.2001 to Dec.2010,36 patients with esophageal carcinoma underwent esophagectomy and descending aortic replacement simultaneously in our hospital.The clinical data were retrospectively reviewed.All patients had a left posterior lateral incision via the 6th intercostal space.The vascular adventitia of the descending thoracic aorta in the left side was incised,and the aorta was clamped in the proximal and distal side of the invaded segment.Then the invaded segment was resected and replaced with artificial vessels.Esophageal carcinoma was radically resceted,and left cervical esophageal-gastro anastomosis was performed in all patients.Results Radical resection of esophageal carcinoma was achieved in all patients.There was no perioperative death,or severe complications such as paraplegina,acute renal failure and intestinal dysfunction occurred.Two patients had chylous hydrothorax,and one had late stage anastomotic stoma fistula.The post-operative hospital stay was 10-42 d,mean (15.5 ± 7.2 )d.The pathological examination revealed that all the patients had squamous cell carcinoma.The aortic tunica adventitia was invaded in all the patients,9(25％ ) had tunica media invasion,and there was no tunica intima invasion observed.The 1-,3-,and 5-year survival rate was 80.6％,46.2％ and 20.0％ respectively.Conclusion Combined esophagectomy and descending aortic replacement for locally advanced T4 esophageal carcinoma invading aorta can be considered as radical operation for selected patients,and it can improve the survival rate and life quality of the patients.

Objective To compare the effect of brain unilateral and bilateral antegrade selective cerebral perfusion (ASCP) during a hybrid approach to DeBakey type Ⅰ aortic dissection and to provide the clinical evidence for the selection of cerebral perfusion methods of aortic dissection surgery.Methods Among the 56 patients undergoing a hybrid approach to DeBakey type Ⅰ aortic dissection from January 2009 to June 2011,24 were enrolled in the study.They were randomly divided into a unilateral ASCP group (n =11) and a bilateral ASCP group (n =13).The patients in both groups underwent cognitive ability test,brain CT scan,cerebrovascular and aortic CTA examinations before and after procedure.The general information,intraoperative conditions and neurological function in the patients of both groups were compared.Results There was no difference in the general data between the unilateral and bilateral ASCP groups.There were no significant differences among the intraoperative cardiopulmonary bypass time (125.2 ± 34.4 min vs.132.1 ± 45.4 min; t =- 0.278,P =0.784),aortic cross-clamping time (54.5 ± 23.6 min vs.61.6 ± 27.5 min; t =-0.149,P =0.883),cerebral perfusion time (30.9 ± 13.2 min vs.31.7 ± 14.5 min; t =- 1.283,P =0.213),right radial artery pressure (57.6 ± 15.5 mm Hg vs.60.7 ± 14.3 mm Hg; t =0.758,P =0.457),and arterial oxygen pressure (465.6 ± 62.4 mm Hg vs.488.4 ± 72.5 mm Hg; t =- 1.894,P =0.071 ).There were no surgery and recent death in both groups.There were no significant differences among the mechanical ventilation time (33.5 ± 14.6 h vs.37.8 ± 12.3; t =- 1.009,P =0.162),time awake after surgery (5.2 ± 2.4 h vs.5.5 ± 3.1 h; t =0.876,P =0.195),and intensive care unit stay time (7.5 ± 3.1 d vs.8.2 ± 3.5 d; t =-0.186,P =0.427).There was no new permanent neurological dysfunction in both groups.One patient had transient neurological impairment in each group.The cognitive function scores after surgery in the unilateral ASCP group (50.1 ± 14.8 vs.47.3 ± 15.2; t =1.005,P =0.126) and in the bilateral ASCP group (52.1 ± 13.7 vs.48.6 ± 16.5; t =0.576,P =0.254) were slightly lower than those before procedure,however,there was no significant difference; there was also no significant difference in the unilateral and bilateral ASCP groups before (t =-0.887,P =0.385) and after procedure (t =-0.953,P =0.351).Conclusions Under the circumstance of complete circle of Willis,the brain protective effect of the unilateral and bilateral ASCP in patients undergoing a hybrid approach for DeBakey type Ⅰ aortic dissection had no significant difference,and more simple and convenient unilateral ASCP can be used.

Objective The present study aims to summarize the clinical experience and methods of nervous system protection in the treatment of DeBakey type Ⅰ aortic dissection with hybrid procedure.Methods From Jan 2009 to Jun 2011,56 patients suffering from DeBakey Ⅰ aortic dissection underwent hybrid procedure.The ascending aorta part were replaced under conventional cardiopulmonary bypass,and the0 aortic arch branch vessels were reconstructed,and then a stent graft was implanted to cover the aortic arch and part of the descending aorta.The unilateral antegrade cerebral perfusion(UACP) and/or bilateral antegrade cerebral perfusion (BACP) combined with femoral artery perfusion was used,and the left subclavian arteries were selectively reconstructed according to cerebral arteries and aorta computed tomography angiography scan.Results All the patients went through the procedure successfully.BACP combined with femoral artery perfusion was applied in 16 patients,UACP combined with femoral artery perfusion in 33 patients,and 7 were perfused with only femoral artery cannulation.Of all the patients,19 underwent the innominate artery and left common carotid artery reconstruction and the other 37 patients underwent the innominate artery,the left common carotid artery and the left subclavian artery reconstruction.The cardiopulmonary time was 44 -95 min,mean (65 ±24) min; aortic clamping time was 32 -71 min,mean (48 ±29)min; the cerebral perfusion time was 24 -44 min,mean (32 ± 13) min.One ( 1.8％ ) patient,who was perfused with only femoral artery cannulation,suffered from permanent neurological dysfunction,and 5 (8.9％) had transient neurological dysfunction.One patient died from severe infection,1 patient was given up because of permanent neurological dysfunction,and the rest 54 patients recovered and discharged.The patients were followed up 1 to 25 months,and there was no newly occurred neurological dysfunc tion.The CTA examination 3 months post-operative revealed that the branch bypass vessels were unobstructed.There was no subclavain steal symptom occurred in the 19 patients whose left subclavian arteries were not reconstructed.The left upper limb strength was slightly lessened in 3 patients and recovered 6 - 12 months later.Conclusion In the treatment of DeBakey type Ⅰ aortic dissection with hybrid procedure,the selective use UACP and/or BACP combined with femoral artery perfusion can avoid deep hypothermic and circulatory arrest and provide the continuous cerebral and spinal perfusion.This perfusion strategy in hybrid procedure can decrease the morbidity of post-operative nervous system disorders with satisfactory nervous system protection effect.

Objective To review the clinical experience in the surgical treatment of infective endocarditis,and to summarize the key points of how to elevate therapeutic effect.Methods From Jan 2001 to Dec 2010,106 patients with infective endocarditis who underwent operative therapy were retrospectively analyzed.All patients underwent cardiac operation in conventional hypothermic cardiopulmonary bypass.Vegetations and suspicious infective focus were thoroughly cleaned.Endocardium was swabbed with normal sodium repeatedly and with high concentration antibiotic solution.Combined anomalies were rectified and the affected valves were replaced.Artificial valves were preconditioned with antibiotics before implantation.All patients were treated with full dose of sensitive antibiotics for 6-8 weeks after operation.Results Two patients(1.8％) died perioperatively,1 died of severe pulmonary infection 5 days post operation,and 1 died of multisystem organ failure.Other 104 patients recovered smoothly,95 of whom were followed up for 6 months to 10 years.One patient died,and the other patients recovered with Ⅰ-Ⅱ grade heart function(NYHA).Three patients had anticoagulation related complications.There was no relapse of endocarditis occurred during the period of follow-up.Conclusion Early diagnosis and timely operation on infective endocarditis can achieve satisfactory effect.The thorough sterilization,prosthetic valves precondition and post operative regular antibiotics therapy were the key points of successful surgical treatment.

Objective Some major procedures for DeBakey type Ⅰ aortic dissection used to be performed with deep hypothermic circulatory arrest, which had been associated with more complications than seen with standard extracorporeal circulation. We reviewed the cases who received the treatment for DeBakey type Ⅰ aortic dissection by hybrid procedure without deep hypothermic circulatory arrest. The procedure consisted of ascending aorta replacement, ascending aorta-aortic arch branch vascular bypass reconstruction and endovascular graft exclusion. Methods From January 2009 to June 2010, 39 patients [mean age (55 ±16) years] who had DeBakey Ⅰ aortic dissection underwent hybrid procedure without deep hypothermic circulatory arrest. The femoral artery and right axillary artery were cannulated for perfusion. The ascending aorta and/or aortic valves were replaced under conventional extracorporeal circulation with Bentall procedure or Wheat procedure. The aortic arch branch vessels were dissected and the proximal part was sealed. Then the ascending aorta-aortic arch branch vascular bypasses were constructed with 4-bifurcation vascular grafts, Y-shape bifurcated vascular grafts or artificial vessels. Finally the endovascular grafts were deployed via the femoral incisions monitored dynamically with DSA, or via the ascending aortic bifurcated vessels monitored with transesophageal echocardiography. Results The operation succeeded in all 39 patients. Eight patients underwent ascending aorta replacement without aortic valve replacement or prosthesis, 20 patients underwent Bentall procedure ( Carbrol procedure were used in 11 cases), and 11 underwent Wheat procedure. For ascending aorta-aortic branch vascular bypass reconstruction, sequential anastomoses were performed in 8, Y-shaped bifurcated grafts were used in 15, and 4-bifurcated grafts were employed in 16 patients. The endovascular stent grafts were deployed via the former femoral incisions in 36 patients and via ascending aortic bifurcated vessels in 3. The cardiopulmonary bypass time was (61 ±22) minutes, the aortic crossclamp time was (48 ±18) minutes, and the post-operative intubation time was (30 ±9) hours. The thoracic drainage from each patient was less than 300 ml in 24 hours. No complication, such as hemiplegia, paraplegia, severe infections, renal failure or coagulation disorder, was observed. The duration of hospitalization was (21 ±6) days. No hospital death occurred. Follow-up was performed 1 to 15 months [mean (8.4 ±7.2) months] postoperatively. All patients survived without any organ dysfunction at follow up. The CTA examination 3 months after operation revealed that the false lumens had been closed in 91.2% of the patients. Conclusion Our findings indicated that the hybrid procedure, which combining ascending aorta replacement,ascending aorta-aortic arch branch vascular bypass reconstruction and endovascular graft exclusion under conventional extracorporeal circulation, may be an option for avoiding the possible complications associated with profound hypothermic circulatory arrest. The novel hybrid operation may improve the surgical outcomes and provide a simplified surgical approach for the treatment of DeBakey Ⅰ aortic dissection.

Objective To observe the effects of Soyasaponins on inflammatory factors, antioxidant activity and exercise ability in rats with severe heat stroke. Methods Eighty male Sprague-Dawley (SD) rats were randomly divided into normal control group, heat shock model group, saline control group and Soyasaponin group, The rats that died during the experiment or with a low rectal temperature (< 41℃) were excluded, and finally 54 rats were included, 18 rats remaining in each group. The rats in the heat shock model group were placed in the simulated hot climate animal cabin at 30 ℃, and the temperature within 30 minutes was raised to 39 ℃ in the cabin with 65% humidity; in the mean time, the rat models of heat shock were replicated under the following situations: let the rats exercise on a treadmill with running speed set at 15 m/min, slope degree 0°, once running for 8 minutes, interval 2 minutes and the heat shock time was 90 minutes, the rats in the normal control group were fed in an environment with temperature ranging from 23-25 ℃ and relative humidity ranging from 50%-70%. After the establishment of models, the saline control group and Soyasaponin group were given daily saline and Soyasaponin (10 mg/kg) respectively by gavage for 3 consecutive months, while the heat shock model group was not given any treatment. The femoral artery blood was collected 24 hours after the rats left the cabin. The serum levels of interleukins (IL-6, IL-1β), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) were measured by enzyme-linked immunosorbent (ELISA) and the contents of serum hemoglobin (Hb), serum urea (BUN), lactate dehydrogenase (LDH) and blood lactic acid (Lac) were measured by automatie biochemical analyzer. Results The levels of IL-6, IL-1β, TNF-α, IFN-γ, MDA, Hb, BUN, LDH, Lac in heat shock model group were significantly higher than those of the normal control group [IL-6 (ng/L): 86.17±4.82 vs. 12.60±3.49, IL-1β (ng/L): 83.00±5.98 vs. 15.70±3.64, TNF-α (ng/L): 72.22±6.93 vs. 13.75±2.69, IFN-γ (ng/L): 36.22±3.02 vs. 7.35±1.60, MDA (nmol/mg): 19.78±4.56 vs. 6.40±1.35, Hb (g/L): 136.22±1.93 vs. 126.75±5.84, BUN (mmol/L):21.06±3.44 vs. 5.65±1.35, LDH (μmoL·s-1·L-1): 9.65±0.83 vs. 2.12±0.17, Lac (mmol/L): 552.56±78.33 vs. 1.32±0.18, all P ＜ 0.05], SOD and GSH-Px were significantly lower than those in normal control group [SOD (kU/L):97.89±10.57 vs. 126.65±11.35, GSH-Px (kU/L): 19.22±2.58 vs. 43.45±4.02]; however, the levels of IL-6, IL-1β, TNF-α, IFN-γ, MDA, BUN, LDH and Lac in Soyasaponin group were significantly lower than those in heat shock model group [IL-6 (ng/L): 45.28±3.54 vs. 86.17±4.82, IL-1β (ng/L): 41.61±2.93 vs. 83.00±5.98, TNF-α (ng/L):37.22±2.46 vs. 72.22±6.93, IFN-γ (ng/L): 19.22±2.60 vs. 36.22±3.02, MDA (nmol/mg): 11.28±1.74 vs. 19.78±4.56, BUN (mmol/L): 11.78±2.13 vs. 21.06±3.44, LDH (μmoL·s-1·L-1): 3.70±0.26 vs. 9.65±0.83, Lac (mmol/L): 274.56±59.08 vs. 552.56±78.33, all P < 0.01], SOD, GSH-Px and Hb were significantly higher than those of heat shock model group [SOD (kU/L): 116.11±11.28 vs. 97.89±10.57, GSH-Px (kU/L): 31.17±2.90 vs. 19.22±2.58, Hb (g/L): 141.33±3.79 vs. 136.22±1.93, all P < 0.01]; there were no significant statistical differences in above indexes between heat shock model group and saline control group (all P > 0.05). Conclusion After heat shock and exercise management, the production and release of inflammatory factors are increased, and the level of lipid peroxidation was elevated in rats. The Soyasaponin can improve the ability to withstand heat shock and strong exercise by reducing the production and release of inflammatory factors and lipid peroxidation in the rats with severe heatstroke.

Objective:To explore the clinical and genetic characteristics of a fetus diagnosed with Congenital myasthenic syndrome type 16 (CMS16).Methods:A couple who had visited Tianjin Medical University General Hospital in February 2018 due to "adverse outcome of two pregnancies" was selected as the study subject. Clinical data was gathered. Peripheral blood and amniotic fluid samples were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Low-depth whole-genome sequencing was carried out to detect copy number variation (CNV) in the fetus.Results:The couple′s first pregnancy had resulted in a miscarriage at 27 + 5 weeks, when ultrasound had revealed pleural effusion and polyhydramnios in the fetus. Their second pregnancy was terminated at 30 + 5 weeks due to fetal hand malformations, polyhydramnios and pleural fluid. Both couple had denied family history of genetic conditions. For their third pregnancy, no CNV abnormality was detected, whilst a compound heterozygous variants, including a maternally derived c. 3172C>T (p.R1058W) and paternal c. 1431delG (p.K477fs*89) in the SCN4A gene were detected. Based on the guidelines from the American College of Medical Genetics and Genomics, the c. 3172C>T (p.R1058W) was predicted as a likely pathogenic variant (PM1+ PM2_supporting+ PP3+ PP4), whilst the c. 1431delG (p.K477fs*89) was predicted as a pathogenic variant (PVS1+ PM2_supporting+ PP4). Conclusion:The c. 3172C>T (p.R1058W) and c. 1431delG (p.K477fs*89) compound heterozygous variants of the SCN4A gene probably underlay the CMS16 in the third fetus.

OBJECTIVETo investigate the effect of temperature on the stability of intermediate and final products of inactivated enterovirus 71 vaccine, which was prepared in human diploid cells.

METHODSThe different batches of harvest viral cultures, the vaccine stock solutions and the final productions of inactivated enterovirus 71 vaccine were stored at different temperatures. The samples of viral culture stored at -20°C or 4°C were harvested at 0, 6, 12 and 24 months later. The samples of vaccine stock solutions stored at -20°C were harvested at 0, 6, 12 and 24 months later, and that stored at 4°C were harvested at 0, 1, 3, 6 and 12 months later. The samples of finial products were harvested at different time points (0, 6, 12 and 24 months for storing at 4°C; 0, 7, 14, 28, 42 and 60 d for storing at 25°C; 0, 3, 7, 14 and 21 d for storing at 37°C). The viral titer, antigen content, antigen purity, endotoxin content, effectiveness, pH and appearance of samples were determined, respectively. A total of 1 800 BLAB/c mice were immunized by vaccine and 150 control mice were injected by diluents without antigen via intraperitoneal. The tail vein blood (500 µl per mouse) from 1 950 mice were harvested after 4 weeks post injected. The neutralization antibody titers of the serum were tested to calculate the half effective dose (ED50) of final products. All results were analyzed using analysis of variance to compare the differences of the above indexes.

RESULTSThe viral titers of harvest viral culture of inactivated EV71 vaccine were (6.67 ± 0.13), (6.56 ± 0.09), (6.52 ± 0.04), (6.39 ± 0.16) lgCCID50/ml (CCID50, the half cell culture infective dose) after 0, 6, 12 and 24 months storage at -20°C; and (6.67 ± 0.13), (6.41 ± 0.13), (6.19 ± 0.18), (5.97 ± 0.09) lgCCID50/ml at 4°C. The viral titers reduced with time (F = 9.81 or 44.16, P < 0.05). The antigen contents of the vaccine stock solution were maintained at (3 626.67 ± 1 382.56) EU/ml within 3 months at 4°C, but were (2 080.00 ± 876.36), (951.17 ± 346.35) EU/ml at 6 and 12 months, respectively. The ED50 of the final production were (31.00 ± 2.71), (32.93 ± 3.22), (39.37 ± 3.44) and (46.04 ± 3.25) EU/ml after 0, 6, 12 and 24 months storage at 4 °C, but were (31.00 ± 2.71), (32.23 ± 2.66), (34.70 ± 1.77), (40.04 ± 2.10), (47.78 ± 1.93) and (56.97 ± 0.50) EU/ml at 0, 7, 14, 28, 42 and 60 days at 25°C, and were (31.00 ± 0.00), (36.20 ± 0.00), (41.87 ± 0.50), (53.25 ± 0.50) and (64.84 ± 0.58) EU/ml at 0, 3, 7, 14 and 21 days at 37°C, respectively. The ED50 had increased with the time by and had significantly differences compared with the beginning level (F = 28.49, 215.15 or 156.12, P < 0.05).

CONCLUSIONThere is a good stability of the intermediate and final productions of inactivated enterovirus 71 (EV71) vaccines, within 24 months at -20°C or 6 months at 4°C storage for viral culture, 24 months at -20°C or 3 months at 4°C storage for stock solution and 24 months at 4°C or 28 d at 25°C or 7 d at 37°C storage for finial vaccine.

Animals ; Drug Storage ; methods ; Enterovirus A, Human ; Humans ; Immunization ; Mice ; Vaccination ; Vaccine Potency ; Vaccines, Inactivated