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Objective:To explore the application of high-throughput sequencing (HTS) technology in pathogens detection for spinal infection.Methods:From January 2019 to May 2020, a total of 41 patients including 31 males and 10 females with an average age of 59.7±11.9 years (29-75 years) were suspected of spinal infections. There were 37 patients with local pain, 15 with fever (≥38 ℃) and 18 with neurological dysfunction. The infected sites were as follows, 4 cases of cervical spine, 8 cases of thoracic spine and 29 cases of lumbar spine. There were 36 patients met the surgical indications and underwent open debridement, bone grafting, fusion and internal fixation, while the other 5 patients underwent conservative treatment (three received drug therapy and two were transferred to the internal department for chemotherapy). Lesions obtained from open surgery patients were underwent pathology and HTS examination. In 5 cases with conservative treatment, two of them underwent CT guided percutaneous puncture for samples, while one case underwent ultrasound guided percutaneous puncture for pus, one case for venous blood, and one case received lumbar puncture for cerebrospinal fluid. The samples were sent for pathological and HTS examination, while liquid specimens were sent for bacterial culture and HTS. The sensitivity and specificity of HTS results were determined according to pathological examination which was regarded as the "gold standard". Based on HTS results combined with the clinical manifestations, imaging examination and pathological results of the patients, targeted antibiotics or anti-tuberculosis drugs were selected for postoperative drug therapy. Patients with bacterial infection received anti-infection treatment for 3 months after operation. For tuberculosis patients, "tetrad" (isoniazid+rifampicin+pyrazinamide+ethambutanol) anti-tuberculosis treatments were underwent for one year. Inflammation indicators from the blood samples were observed before and after treatment, including white blood cell count (WBC), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). These indicators were used to monitor disease progression and the curative effects. All patients were followed up for at least 3 months after surgery.Results:A total of 41 patients with suspected spinal infection were included in this study. The HTS pathogen detection results were obtained within 48 h. For the initial 5 patients, first-generation sequencing verification was conducted with coincidence rate 100%. Further, no further verification was conducted in the rest patients. Among the 41 cases, a total of 26 cases had positive results with a positive rate of 63.4%(26/41). Among them, thirteen cases were with mycobacterium tuberculosis (31.7%) and 6 cases with staphylococcus (14.6%). Fungi and Brucellosis were diagnosed in 2 cases respectively, accounting for 4.9% respectively. The test were negative in 15 patients (36.6%), including 2 patients with tumor or tumor-like lesions (1 hematologic tumor and 1 eosinophilic granuloma). A total of 38 patients underwent pathological examination, which confirmed 7 cases of suppurative infection, 12 cases of tuberculosis, 2 cases of tumor or tumor-like lesions and the remaining 17 cases of inflammatory lesions. The sensitivity and specificity of HTS were 80%(16/20) and 55.6% (10/18) with positive predictive value (PPV) 66.7% (16/24) and negative predictive value (NPV) 71.4% (10/14). All patients were followed up for 3 months. The inflammation indicators of blood at 3 months were all lower than that at admission. WBC decreased from (7.50±3.26)×10 9/L at admission to (6.22±2.53)×10 9/L at 3 months after treatment without statistically significant difference ( t=1.082, P=0.290). The CRP decreased from (32.2±34.1) mg/L to (4.5±10.5) mg/L, and ESR from (44.2±26.5) mm/1 h to (18.6±12.1) mm/1 h with statistically significant difference ( t=8.963, P<0.001; t=5.421, P<0.001). Conclusion:High-throughput sequencing technology can be used in detection of spinal infection pathogens, due to its relatively high positive rate, satisfied sensitivity and good diagnostic value.
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Objective To investigate the expression of NMDAR1 in the hippocampus and cerebral cortex of old rats after 30-min -inhalation of 2% isoflurane, and to investigate the effects of isoflurane on the learning and memory functions of old rats and the underlying mechanism. Methods The healthy old male Sprague Dawley rats (n = 36) were randomly divided into the control group, the oxygen group, the 2-hour post-recovery group, the 1-day post-recovery group, the 3-day post-recovery group, and the 7-day post-recovery group. The morris water maze was used to detect the ethological effect of 30-min inhalation of isoflurane , and the immunohistochemistry assay was used to detect the expression of NMDAR1 in the hippocampus (CA1, CA3) and the cerebral cortex. Results The 30-min inhalation of 2% isoflurane inhibited the learning and memory abilities of old rats at 2 h post-recovery. On 1 d post-recovery, the inhibition of learning and memory began to reduce, then on 3 d and 7 d post-recovery, the learning and memory abilities continously recovered. The expression of NMDAR1 in the rat hippocampus and cerebral cortex decreased at 2 h post-recovery, and reversed on 1 d post-recovery and reached the normal level on 3 d and 7 d post-recovery. Conclusion 30-min inhalation of 2%isoflurane had an inhibitory effect on the learning and memory abilities of old rats, and the attenuation of NMDAR1 in the hippocampus and cerebral cortex may involve in this process.
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Objective To investigate the effect of different volume of fresh gas on postoperative analepsia period of general anesthesia with sevoflurane in patients undergoing abdominal surgery. Methods 181 ASAⅠ-Ⅱpatients aged 20 - 70 undergoing elective abdominal surgery were divided into group A (112 patients) and group B (69 patients). Both groups received sevoflurane general anesthesia during surgery. At the end of surgery , group A received fresh gas for no more than 2L/min , and group B received the gas for more than 2L/min. The recovery time of spontaneous breathing, eye-opening time (from the end of surger to eye opening), and time of endotracheal extubation (from the end of surgery to endotracheal extubation ) were recorded. Results There were no significant differences in the recovery time of spontaneous breathing ,eye-opening time and time of endotracheal extubation between group A and group B (P > 0.05); Gender and time of intraoperative continuous uses of sevoflurane had effects on the recovery time of spontaneous breathing , which showed statistical significances (P < 0.05). Conclusions Uses of fresh gas for no more than 2L/min or more than 2L/min after surgery have no marked effects on the recovery time. However , gender and time of intraoperative continuous uses of sevoflurane may have effects on the recovery time of spontaneous breathing.
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Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients.Methods Twenty ASA Ⅰ or Ⅱ patients (aged 20-44 years and weighing 40-75 kg) requiring blood salvage during operation (with the estimated intraoperative blood loss of 15-20 ml/kg) were randomly divided into two groups (n =10 each):control group (group C) and leukocyte depletion group (group D).Anesthesia was induced with midazolam,fentanyl,propofol and vecuronium and maintained with isoflurane inhalation and intravenous infusion of propofol and remifentanil.The patients were mechanically ventilated after endotracheal intubation.Intraoperative blood salvage and reinfusion were performed in all the patients.In group D,the salvaged blood was filtered by a leukocyte depletion filter placed in the line of the reinfusion circuit.Blood samples were collected from the central vein before anesthesia (T1),at the end of surgery (T2) and 12 hours (T3) and 36 hours (T4) after operation in all the patients.The rosette rates of red blood cell-C3b receptors (RBC-C3bRR) and RBC-immune complex (RBC-ICR) were determined.The number of leukocytes and polymorphonuclear neutrophils (PMNs) were counted.The plasma levels of interieukin-6 (IL-6),tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured,too.Results Compared with group C,the plasma levels of IL-6,TNF-α and MDA were significantly lower at T2-T4,the RBC-C3bRR was significantly higher at T3-T4 (P ＜ 0.01),and the number of PMNs was significantly lower at T4 in group D(P＜0.05).Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients,and the decrease of systemic inflammatory response may be involved in the mechanism.
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ObjectiveTo investgate the effects of pyruvate sodium on energy metabolism of erythrocytes in dog blood during circulation in cardiopulmonary bypass machine.MethodsTen healthy adult mongrel dogs of both sexes weighing 12-18 kg were anesthetized with intraperitoneal 2.5％ pentobarbital 25 mg/kg.Femoral artery was cannulated and 500 ml of blood was collected from femoral artery in each dog.Blood samples from the 10 dogs were divided into 2 equal parts (250 ml each).Each part was mixed with 250 ml lactated Ringer's solution (group C) or 250 ml pyruvate sodium (group P).Flow rate of 4 L/min was employed and blood temperature was kept at 30-32 ℃ during circulation in cardiopulmonary bypass machine.The blood sanples were collected before (baseline value,T1 ) and at 30,60,90,120 min of circulation (T2-5) for determination of ATP,ADP and AMP concentrations in RBC.ResultsThe ATP and AMP concentrations were significantly higher at T2-5,while ADP concentration was higher in group P than in group C.ConclusionPyruvate sodium can improve energy metabolism of erythrocytes in dog blood druing circulation in cardiopulmary bypass machine.
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Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients. Methods Twenty ASA Ⅰ or Ⅱ patients, aged 20-44 yr, weighing 40-75 kg, required blood salvage during operations (intraoperative blood loss expected 15-20 ml/kg), were randomly divided into 2 groups ( n = 10 each): control group (group C) and leukocyte depletion group (group D) . Anesthesia was induced with midazolam, fentanyl, propofol and vecuronium and maintained with isoflurane inhalation and iv infusion of propofol and remifentanil. The patients were mechanically ventilated after tracheal intubation. Intraoperative blood salvage and reinfusion were performed in all patients using cell saver system. In group D, the salvaged blood was filtered with a leukocyte depleting filter placed in the line of the reinfusion circuit. Blood samples were collected from the central vein before anesthesia (T1 ) , at the end of surgery (T2 ) , and at 12 h (T3 ) and 36 h (T4 ) after operation in the two groups. The rosette rates of RBC-C3b receptors and RBC-immune complex were determined. The leukocyte and neutrophil were counted. The plasma levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured. Results The plasma levels of IL-6, TNF-a and MDA were significantly lower at T2-4 , the rosette rate of RBC-C3b receptors was significantly higher at T3-4 ( P ＜ 0.01) , and the neutrophil count was significantly lower at T4 in group D than in group C ( P ＜ 0.05) . Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients, and the decrease in the systemic inflammatory response may be involved in the mechanism.