Objective:To explore the epileptic phenotype spectrum of SLC6A1 gene mutations and their genotype-phenotype correlation. Methods:Four hundred patients with epilepsy of unknown etiology admitted to Epilepsy Center, Department of Neurology, Second Affiliated Hospital of Guangzhou Medical University from July 2019 to July 2024 were enrolled to screen the SLC6A1 gene mutations; the clinical characteristics, mutation pathogenicity, and changes of hydrogen bond between amino acids, stability and amino acid hydrophobicity of SLC6A1 gene encoded proteins caused by missense mutations in patients with SLC6A1 gene mutations were analyzed. At the same time, a comprehensive search was conducted in PubMed, HGMD and CNKI databases to collect the publicly reported SLC6A1 gene mutations related to epilepsy up to September 8, 2024; differences in proportion of missense mutations between the two most common and featured epileptic phenotypes and proportion of missense mutations in loops of SLC6A1 gene coding proteins were analyzed. Results:Five patients carried SLC6A1 gene mutations in 400 patients with epilepsy of unknown etiology: 2 had de novo heterozygous canonical splice site mutations (c.850-1G>A and c.1324-1G>A), with phenotypes as partial epilepsy combined with severe development delay and childhood absence epilepsy combined with mild developmental delay; 2 had de novo heterozygous missense mutations (c.187G>A/p.Gly63Ser and c.1081C>A/p.Pro361Thr), with phenotypes as partial epilepsy combined with mild development delay and generalized epilepsy combined with severe development delay; and one had heterozygous missense mutation of unknown origin (c.700G>A/p.Gly234Ser), with phenotype as Lennox-Gastaut syndrome. Four de novo mutations were evaluated as having pathogenic or likely pathogenic features, and one mutation of unknown origin was evaluated as of uncertain significance. In addition, 3 missense mutations caused significant changes in number or bonding form of hydrogen bonds between amino acids of the encoded proteins, with obviously decreased stability and hydrophobicity of the encoded proteins. (2) Results of literature analysis showed that 84 SLC6A1 mutations have been reported to be associated with epilepsy; combined with the genetic results in this study, a total of 89 SLC6A1 mutations were identified, including 53 missense mutations, 33 nonsense mutations, and 3 in-frame/in-del mutations; 7 epilepsy phenotypes were involved, including 38 patients with myoclonic atonic epilepsy (MAE), 16 with epilepsy, 12 with epileptic encephalopathy, 8 with childhood absence epilepsy, 6 with childhood-onset epilepsy, 6 with generalized epilepsy, and 3 with focal epilepsy. No significant difference in proportion of missense mutations was noted between MAE and epileptic encephalopathy patients ( P>0.05); however, the proportion of missense mutations in loops of the epileptic encephalopathy patients was significantly higher than that of the MAE patients ( P<0.05). Conclusion:SLC6A1 gene mutations can cause complex and diverse epilepsy phenotype spectrum, and most patients are accompanied by developmental delay; subregional effect of the encoded protein molecules may be a potential mechanism for different clinical phenotypes between MAE and epileptic encephalopathy caused by SLC6A1 gene mutations.

Objective To evaluate the surgical treatments of refractory sclerosing peritonitis related peritoneal dialysis.Methods Clinical data of 15 patients with refractory sclerosing peritonitis related to peritoneal dialysis treated in the General Surgery Department of the 900th Hospital of the Joint Logistics Support Force of the People's Liberation Army from June 30,2014 to May 30,2018.Among them,5 cases underwent"open abdomen peritoneal catheter removal+intestinal adhesiolysis+abdominal infection flushing and drainage with catheter",4 cases underwent"laparoscopic peritoneal catheter removal+intestinal adhesiolysis+abdominal infection flushing and drainage with catheter",3 cases underwent"laparoscopic peritoneal dialysis catheter removal+abdominal infection flushing and drainage with catheter",2 cases underwent"open abdomen peritoneal dialysis catheter removal+abdominal infection flushing and drainage with catheter",and 1 case underwent"laparoscopic examination combined with laparotomy exploration and removal of lower abdominal catheter+intestinal adhesiolysis+abdominal infection flushing and drainage with catheter".Age,gender,clinical symptoms,abdominal CT examination,peripheral blood routine,blood biochemistry,blood C-reactive protein(CRP),white blood cells,biochemistry,and aetiology of peritoneal dialysis fluid were collected and followed up,and the therapeutic effect was evaluated.Results 15 patients were transferred to the Department of Surgery after ineffective treatment in the Department of Internal Medicine.Preoperatively(after 5 days of antibiotic treatment)compared to before antibiotic treatment,there were no significant changes in blood WBC,blood NEUT％,CRP,and peritoneal fluid WBC(P＞0.05).Laparoscopic exploration or laparotomy exploration was performed,during which the peritoneal dialysis catheter was removed and the abdominal infection focus was cleared.A pelvic cavity washout drainage tube was left in place postoperatively.Fourteen patients had a good recovery after surgery,with effective control of peritonitis symptoms and no complications such as intestinal obstruction or enterocutaneous fistula.After the removal of the peritoneal dialysis catheter,all patients switched to hemodialysis.A comparison of inflammatory markers before and after surgery showed a significant decrease after surgery.Three days postoperatively compared to before surgery(after 5 days of antibiotic treatment),there were no significant changes in blood WBC,blood NEUT％,CRP,and peritoneal fluid WBC(P＞0.05).Seven days postoperatively compared to before surgery(after 5 days of antibiotic treatment),there was a significant decrease in blood WBC[(7.43±2.65)× 109/L VS(10.17±5.24)× 109/L],blood NEUT％[(88.23±9.02)％VS(85.07±11.57)％],and CRP[(152.88±113.01)mg/L VS(114.49±92.97)mg/L](P＜0.05);the peritoneal fluid WBC at 7 days postoperatively showed no significant change compared to before surgery(after 5 days of antibiotic treatment)(P＞0.05).The cases were followed up for at least 22 months,and 13 patients did not experience peritonitis or intestinal obstruction again.One patient died 39 days after surgery due to multiple organ failure,and one patient died from other causes after a 2-year follow-up.Conclusion For refractory sclerosing peritonitis related peritoneal dialysis that is ineffective in medical conservative treatment,On the basis of reasonable and effective antibiotics to control infection,surgical intervention should be actively carried out and surgical methods such as surgery should be used to control the progress of peritonitis,reduce mortality and improve the cure rate.

Objective:This paper introduces the concepts of " patient-centered cell therapy research management model", to provide reference for domestic medical institutions which conduct cell therapy clinical trials.Methods:We reviewed and summarized the experiences of conducting cell therapy at Peking University Shenzhen Hospital, including challenges and response plans regarding to the technology assessment, ethical evaluation risks and regulatory compliance. According to which, this paper aims to explore the reflections and practical experience of establishing a patient-centered, multi-stakeholder shared decision-making research management system.Results:The " patient-centered cell therapy research management model" ensures the reliability of research results through multi-stakeholder engagement in decision-making and management, adequate technical evaluation, effective ethical review and harmonized scientific research management, which not only meets the urgent health needs of the patient, but also promotes the standardized development of emerging technologies.Conclusions:The " patient-centered cell therapy research management model" is tailored for the cell therapy research, it is important to promote its further assessment and applications.

Objective:To investigate the effect of artificial dermis combined with KCI negative pressure suction device in the repair of scalp defect with skull exposure after the operation of scalp malignancy.Methods:From January 2016 to June 2018, 18 patients with scalp defect and skull exposure after scalp malignant tumor surgery were treated with artificial dermis combined with KCI negative pressure suction device.Results:Eighten patients had good wound healing after the first stage of surgery, and no infection occurred. The time for artificial dermis to complete granulation was 10-14 days, with an average of 12.1 days. After the second stage of skin grafting, the skin graft had a survival rate of 100%, with uniform color and good elasticity, without obvious scar hyperplasia or contracture. The postoperative follow-up period was 6 to 2 months, all the patients healed well without tumor recurrence, with good skin elasticity and friction-resistant, and they were satisfied.Conclusions:Artificial dermis combined with KCI negative pressure suction device is an effective and simple method to repair the scalp defect with skull exposure after tumor surgery.

Objective To investigate the effects of adoptive reinfusion of regulatory T cell (Treg) on the recovery of islet function and graft survival time after islet allograft transplantation. Methods The diabetic model was established using C57BL/6 mice as recipients, and Balb/c mice were chosen as donors for islet allografts transplantation beneath the renal capsule. The recipient mice were divided into 3 groups and 3 mice in each group according to different processing Methods: Treg experiment group (Treg group, 1×10⁶ Treg cells were injected via tail vein at 1 d before operation), positive control group [sirolimus (SRL) group, SRL at a dose of 300 μg/(kg·d) was intragastrically given every day from 1 d before operation] and blank control group (control group, an equivalent volume of normal saline was intragastrically given every day from 1 d before operation). Enzyme-linked immune absorbent assay (ELISA) was used to detect the changes of blood glucose and C-peptide in mice within 14 days after transplantation. In vivo imaging technique was used to dynamically monitor the survival of mice within 14 days after transplantation. Results In each group, the blood glucose levels at postoperative 3 d were significantly decreased compared with those before transplantation (all P < 0.001). At postoperative 1 d, the C-peptide levels showed an explosive rise to varying degree in each group. At postoperative 3 d, the C-peptide levels in each group were significantly higher than that before operation (all P < 0.001). At the end of the observation period at 14 d after operation, the C-peptide levels in the SRL and Treg groups were (427±50) pmol/L and (833±57) pmol/L, relatively higher than that in the control group. But the blood glucose levels were (14.5±0.5) mmol/L and (12.1±0.6) mmol/L, significantly lower than that in the control group (all P < 0.001). Compared with the SRL group, the explosive release amount of C-peptide was significantly lower, the declining trend was more remarkably stable, and the C-peptide level was considerably higher in the Treg group at the end of the observation period (all P < 0.001). At postoperative 14 d, the grafts were almost completely apoptotic in the control group, over 50% of the grafts survived in the SRL group, and over 80% of the grafts survived in the Treg group. Conclusions Adoptive reinfusion of Treg cells can effectively protect islet grafts, prolong the survival time of grafts, and maintain the normal levels of blood glucose and C-peptide in the recipient mice.

Objective To explore the surgical procedures and effects for recurrent scalp dermatofibrosarcoma protuberans (DFSP).Methods Retrospective review was conducted in 7 scalp recurrent DFSP cases in the past 3 years.Intraoperative frozen section analysis of surgical margins was performed.Wide local excision with margins of 3 cm or more was applied,and excision of the skull external lamina or the entire layer of the cranial bones was performed once the skulls under the tumors were invaded.Frozen section analysis was again performed on all the margins.Secondary wound was repaired with local skin flap or fascial pedicled flaps.Surgically removed tumor was paraffin-embedded for HE-staining and immunohistochemical analysis.Results All of 7 tumor specimens were proved recurrent DFSP with histologic findings of intraoperation or postoperation.7 patients were all performed with wide local excision with margins of 3 cm or more,also with skull excision (5 cases with excision of skull external lamina and 2 cases with excision of the entire layer of skull).Negative margins were confirmed with intraoperative frozen sections or postoperative paraffin-embedded sections analysis were confirmed.Immunohistochemical analysis demonstrated that Ki-67 was all positive and CD34 was partially positive in 7 cases.No tumor recurrence or metastasis had been observed in any of our patients after a median follow-up duration of 17 months (range 15-41).Conclusions Wide local excision together with the skull invaded by tumors completely is effective surgical treatments for recurrent scalp DFSP; Use of scalp transferring technique could improve wound repair after complete tumor excision.

Objective To study the prevention effects of AduoLa Fuzhenglin(ADL)Oll the brain injury induced by microwave radiation in rats.Methods A total of 140 male Wismr rats were divided randomly into 5 groups,including control group,microwave exposed group,low dosage(0.75 g·kg-1·d-1)group.middle dosage(1.5 g·kg-1·d-1)group and high dosage(3 g·kg-1·d-1)group.Rats in three ADL groups were lavaged with ADL per day for 2 weeks before radiation.After administration,rats were exposed to microwave at 30 mW/cm2 for 15 min.The abilities of learning and memory were detected by Morris water maze,and the contents of amino acids neurotransmitter of hippocampus were detected by HPLC, then the histology and uhrastrncture of hippocampus were observed with light and electron microscope at 6 h,7 and 14 d after exposure.Results The abilities of learning and memory were declined(F=0.000-0.043,P<0.05)from 6 h to 7 d after exposure,and the contents of four kinds of amino acid neurotransmitter in hippocampus were decreased,of which GLU,GLY and GABA were decreased significantly(F=0.000-0.007,P<0.01)at 6h after exposure,then tissue edema,neuronal degeneration,neuron mitoehondria swelling and cavitation,endocytoplasmie rotieulum broaden,synaptic cleft blurred,and perivascular space widen were found in the hippocampus at 6 h and 7 d after exposure.The changes in low dosage group were similar to those of the radiation group.However,in middle and high dosage groups,the abilities of learning and memory were normal to some extent with the significant differences compared to the radiation group from 6 h to 7 d after exposure(F=0.015-0.028.P<0.05).The contents of four kinds of amino acid neurotransmitter were not decreased,especially GLU contents close tO normal level.There were significant differences between middle and high dosage groups and radiation group at 6 h after exposure(F=0.000-0.042,P<0.05).Moreover,no obvious injury in the hippocampus was observed in middle and high dosage groups at 6 h and 7 d after exposure.Conclusions Exposure to 30 mW/cm2 microwave radiation could decrease the abilities of learning and memory,induce amino acid neurotransmitter turbulence,and injure the histology and uhrastructure of hippocampus.ADL at the dosages of 1.5 and 3 g·kg-1·d-1 would have preventive effects on the injury induced by microwave exposure.The concentration of 1.5 g·kg-1 ·d-1 of ADL might be the effective dosage to prevent the brain damage after microwave exposure.

BACKGROUND: Studies have demonstrated that intervention material retention for a period of time in vivo causes pathogenic bacteria surface attachment, proliferation even forming biofilm to become potential source of high polymer catheter-associated infection. Inhibition of biofilm formation significantly reduces infection.OBJECTIVE: To evaluate the antimicrobial effect of anti-infective polyethylene material and its inhibiting effect to biofilm on the surface.DESIGN, TIME AND SETTING: Single sample observation was performed at the National Engineering Research Center for Engineering Plastics, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences from June 2008 to May 2009, and the antimicrobial test was performed at the General Hospital of General Staff Headquarters of PLA in March 2009.MATERIALS: Organic antibacterial agent2,4,4'-trichloro-2'-hydroxydiphenyl ether; IONPURE~(R) Silver-Inorganic anti-microbialreagent; low-density polyethylene (LDPE).METHODS: LDPE was respectively blended with organic antibacterial agent and inorganic antibacterial agent, and the LDPE slice was prepared by injection molding process.MAIN OUTCOME MEASURES: The antimicrobial efficacy of anti-infective polyethylene was determined by the plate counting method. The ultrasonic-plate method and scanning electron microscopy were introduced to observe the biofilm formation.biofilm maturation. The propagation of microbe was found to be depressed remarkably on the surfaces of modified samples with organic antibacterial agent, and the formation of biofilm was prevented. The observation of SEM confirmed that no biofilm was formed on the surfaces of PE added organic antibacterial agent. The results showed that a large amount of bacteria and extracellularpolysaccharide matrix adhered on the surfaces of PE added inorganic antibacterial agent and untreated PE.CONCLUSION: Anti-infective polyethylene added organic antibacterial agent possesses excellent antimicrobial performance against S. aureus and E coli, and it can inhibit the formation of biofilm on the surfaces, which provides a promising approach to prevent catheter-related infections.

OBJECTIVE: To explore the effects of c-Met-siRNA on the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells in vitro. METHOD: Firstly, the pSilencer 2.0/c-Met-shRNA recombinant plasmid was transfected into laryngeal carcinoma Hep-2 cells with transfecting agent of cationic liposome Lipofectamine 2000. Secondly,the transfection efficacy was tested by RT-PCR and Western-Blot, then the most inhibitive c-Met-siRNA sequence was elected. Cell proliferation, movement and invasion were detected with MTT, cell migration assay and cell invasion assay, respectively. RESULT: After the transfection of pSilencer 2.0/c-Met-shRNA recombinant plasmid into laryngeal carcinoma Hep-2 cells, the expression of mRNA and protein of c-Met decreased significantly in Hep-2 cells, and ability of the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells were also inhibited. CONCLUSION The results indicated that c-Met-siRNA can down-regulated the expression of c-Met and markedly inhibited laryngeal carcinoma Hep-2 cell proliferation, movement and invasion. It may have the potential as a therapeutic modality to treat human laryngeal carcinoma.
Apoptosis ; genetics ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Liposomes ; Proto-Oncogene Proteins c-met ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection