ObjectiveTo study the efficient expression conditions, purification, and partial enzymatic properties of His-tagged recombinant Candida utilis uricase.MethodsThe effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) and lactose as inducers which were added in the end logarithmic phase were compared by the method of shake flask culture. The induction culturing time was studied with 50 L fermentor. The protein of interest was purified by Ni-Sepharose and Sephacryl S-200 HR chromatographies, and the optimal pH value, temperature, and thermal stability were also studied.ResultsThe shake flask culturing experiment results showed that IPTG was better than lactose as an inducer. In the fermentor culturing and at the end of the logarithmic growth stage, the enzyme activity of 164 U per gram bacteria and biomass of 23 grams per litter fermented solution were maximal after adding lactose for 7 hours as an inducer, i.e. the enzyme activity could be collected at 3 772 U per liter. The specific activity of purified uricase was 4.5 U/mg and the optimal pH value and temperature were 7.5 and 40℃. Additionally, the enzyme was stable at pH 6.0–10.0 and the thermal stability was below 45℃.ConclusionsIt is better to use lactose as an inducer in the process of culture. The recombinant uricase with His label can be purified by Ni-column affinity chromatography and Sephacryl S-200HR molecular sieve chromatography, and the purification process is easier than ever before. The optimum temperature, pH value, acid-base stability and thermal stability of the purified enzyme have been established to provide some experimental basis for the relationship between structure and function and practical application in the future.
ObjectiveTo explore the clinical manifestation, diagnosis, treatment and prognosis of infantile spasm complicated with craniostenosis.MethodsA case of infantile spasm complicated with craniostenosis in the Department of Neurology of Qilu Children's Hospital in December 2017 was reviewed with the literature. The clinical manifestations, diagnosis, treatment and prognosis of infantile spasm with craniostenosis were analyzed.ResultsThe proband infantile spasms and craniostenosis was diagnosed by clinical, imaging examination and VEEG. Epileptic attack was prevented and craniostenosis was corrected by hormone shock therapy (corticotrophin was administered for 14 days, followed by topiramate)and surgical treatment (cranial cap reconstruction was performed), and good clinical prognosis was obtained.ConclusionThis case was the first reported case of craniostenosis with infantile spasm in China, and compared with the foreign treatment method, better treatment method and the operation opportunity were obtained. Which has a significant effect on the clinical treatment of infantile spasm complicated with transcranial disease.
ObjectiveTo investigate the efficacy and safety of Stereotactic electroencephalogram (SEEG)-guided Radiofrequency-thermocoagulation (RF-TC) in the treatment of refractory insular epilepsy in children.MethodsThe clinical data of 7 children with SEEG-confirmed insular epilepsy admitted to the Epilepsy Center of the Children’s Hospital Affiliated to Shandong University from January 2021 to May 2022, were retrospectively analyzed (3 males and 4 females; average age, 6.6±3.5 years). All patients underwent stage I pre-operative evaluation, and were implanted with SEEG electrodes for video EEG monitoring. The radiofrequency thermocoagulation contacts were determined according to SEEG and imaging results, and radiofrequency thermocoagulation was performed via electrode contacts. The patients were followed up at 3, 6, 12 and 18 months after operation by outpatient review or via telephone interview. The clinical efficacy was evaluated by Engel classification and complications were recorded. ResultsSix cases (6/7) were characterized by nocturnal seizures, and four cases (4/7) exhibited hypermotor or complex motor seizures. Three cases (3/7) showed focal ankylosis; only 1 patient had aura. All of the 7 cases showed interictal scalp EEG consistent with the side of surgery: 6 cases showed distribution in the perilateral fissure region, and 1 case showed confinement to the temporal region. In MRI, 4 cases showed negative signal, 2 cases showed unclear gray-white matter boundary, and 1 case showed thickening of the insular cortex. All of the 7 patients received electrode implantation and completed follow-up for over 6 months [6.0~22.0 (12.3±5.3) months]. At the last follow-up, 5 of the 7 children were seizure free (Engel class la), and 2 still had seizures after surgery, with no postoperative long-term complications.ConclusionChildren with insular epilepsy rarely show an aura, but have prominent motor symptoms, and the scalp electroencephalogram is mainly distributed in the perilateral fissured area. SEEG-guided RF-TC has good safety and efficacy in the treatment of drug‐resistant insular epilepsy.