Amphetamine-type stimulants ( ATS ) , a group of new-type synthetic drugs mainly in psychological dependence, are abused more and more severely in recent years. MicroRNAs ( MiRNAs ) are an important class of endogenous non-coding small RNAs that mediate posttranscriptional negatively regulation of gene expression by targeting specific mRNA sequences to in-hibit the translation of mRNAs or degrade the expression of mR-NAs. ATS can induce the changes in the expression of miRNAs in addiction-related brain regions which directly involve in the regulation of ATS-induced addictive behaviors. Therefore, to study the regulatory role of miRNAs in ATS-induced addiction has important implications for dependent mechanisms of new-type drugs and the discovery of the new targets of drug actions.

Objective To compare the effects of propofol,nidazolarm versus etomidate combined with sufentanil for anesthesia induction on intraocular pressure.Methods Forty-five ASA Ⅰ or Ⅱ patients,aged 20-40 yr,scheduled for surgery under general surgery,were randomly divided into 3 groups(n =15 each):propofol group(group P); midazolam group(group M)and etomidate group(group E).Anesthesia was induced with iv injection of propofol 2 mg/kg,midazolam 0.2 mg/kg,and etomidate 0.3 mg/kg in P,M and E groups respectively,and then with iv injection of sufentanil 0.2 μg/kg and cisatracurium 0.2 mg/kg in all the groups.The patients were then tracheal intubated.Intraocular pressure(IOP)and MAP were recorded at 1 m in before induction of anesthesia (T0),before intubation(T1),and at 0,1 and 2 min after intubation(T2-4).Results Compared with group P,the incidence of intraocujar hypotension was significantly decreased in group M(P ＜ 0.01).Compared with group E,the incidence of intraocular hypertension was significantly decreased in P and M groups(P ＜ 0.05),The correlation coefficient between MAP and IOP was 0.831,0.889 or 0.806 in group P,M or E respectively(P ＜0.05),and there was no significant difference in the correlation coefficient among the three groups(P ＞ 0.05).Conclusion Midazolam combined with sufentanil for anesthesia induction exerts less influence on lOP and the degree of MAP fluctuations is a major factor contributing to the change in IOP.

Alemtuzumab (Campath) was successfully injected in 21 kidney transplant patients,7 islet transplant patients and 1 simultaneous kidney and islet transplant patient for either prevention or treatment of graft rejection.Prophylactic administration was successfully completed in all patients without discontinuation.Adverse events were not observed in 11 patients (38%),but hypertension in 18 patients (62%),shivering in 3 patients (10.3%),high fever in 3 patients (10.3%),and bronchospasm in 1 patient (3%),respectively.All complications alleviated after proper therapy.During the prophylactic administration of alemtuzumab,strict,timely and proper ward-management was needed.Care for lung,perineum,skin,diet and psychological nursing were necessary.Neither graft acute rejection nor graft chronic rejection episode occurred in all patients during 6 months to 2 years follow-up.Therefore,long term effects of Alamtuzumab and consequences of lymphocytopenia need further observation.

Objective To evaluate the feasibility of methylene blue as a method in lymphatic mapping and the effect factors of SLNB identification rate. Methods The results of SLNB in 276 patients with clinical stage T-T2N0M0 breast eaneer were analyzed to evaluate the effect factors of identification rate and false-neg-ative rate of SLNB. Results The identification of SLN was carried out using methylene blne . When the mapping time of SLN and lymphatic were controlled in 20～30 minutes, the SLN could be visualized easily. SLN was successfully identified in 246 of 276 patients (89.1%) ,423 SLNs were retrieved(1～4). The SLN accurately predicted the status of the axilla in 226 of 246 patients (77.3%) yielding a FNR(false negative rate)of 8.1% (20/246) ,false positive rate of0, and an accuracy of 91.9% (226/241). For the identified rate of SLN, the clinical stage of T2N0M0 disease was higher than the clinical stage of T1N0M0 disease (P=0.046,P<0.05), Upper outer quadrant and lower outer quadrant were higher than other evidently (P<0.0010). SLNs were easily retrieved in those cases aged above 50 years than below 50 years (P<0.001).In the view of false-negative rate of SLNB, the FNR rate in patients of 50 years old was less than older ones (P=0.037, P<0.05). No significant correlation was found between the stage of TMN, the expression of ER and PR, histologic status and FNR. Conclusion When SLNB was performed using methylene blue, we found age, clinical stage of TMN, the location of tumor had relationship with the identified rate of SLN. Ag-ing and location of tumor may adversely affect FNR of SLNB.

BACKGROUND:hIL-24, a tumor suppressor gene, can stimulate immune responses, inhibit the growth of tumor cel s, and the formation of tumor vessels, and induce cel apoptosis. OBJECTIVE:To explore the effects of hIL-24 gene on the proliferation and apoptosis of fibroblasts in the keloid and the underlying mechanisms. METHODS:Al the keloid specimens col ected from 13 patients were used for fibroblast culture and indentification. Fibroblast of the keloid was transfected with or without hlL-24 lentivirus. Subsequently, mRNA expressions of transforming growth factor-β, Smad3, proliferating cel nuclear antigen, matrix metal oproteinase-2,-9, and metal opeptidase inhibitor 1 were determined. RESULTS AND CONCLUSION:Immunofluorescent staining and flow cytometry showed that vimentin antibody was expressed positively in cytoplasma of fibroblast cultures, and the purity was more than 97.8%. Western blot assay showed that hIL-24 expression was significantly increased in the transfected fibroblasts. Quantitative PCR showed that the overexpression rate of hIL-24 in fibroblasts was 81.7%and mRNA expressions of transforming growth factor-β, Smad3, proliferating cel nuclear antigen, matrix metal oproteinase-2, and-9 were significantly decreased, while metal opeptidase inhibitor 1 mRNA expression was significantly increased in hIL-24 transfection group compared with control group (P<0.05). These findings suggest that hIL-24 gene inhibits the expressions of proliferating cel nuclear antigen, matrix metal oproteinase-2, and-9 in fibroblasts, and the underlying mechanism may involves TGF-β/Smad3 pathway.

Objective To screen and identify B cell epitopes in SAG1, SAG2, SAG3, GRA1, GRA6 and P35 antigens of Toxoplasma gondii. Methods The indexes such as hydrophilicity, accessibility, flexibility, secondary structure and polarity of the 6 antigen moleculars above mentioned were analyzed by BioSun system. Two B cell epitopes with high antigenicity from each antigen molecular were selected, and the total twelve pairs of oligonucleotide chains were designed according to the 12 B cell epitopes’ sequence and synthesized, then cloned into plasmid pET-32c. The 12 fragment B cell epitopes were expressed and the expressed fusion proteins were identified with Western blot. Results Twelve B cell epitopes from 6 Toxoplasma antigens (two from each antigen) were predicted and selected. The epitope genes were successfully cloned into pET-32c and expressed. Western blot results showed that 3 of 12 expressed fusion proteins could be recognized by the immunized rabbit sera with soluble antigen of Toxoplasma gondii, but not by the unimmunized rabbit sera Conclusion Three B cell epitopes of Toxoplasma[with potential diagnostic value are obtained.

Objective To analyze the clinical characteristics and pyridox (am)ine-5'-phosphate oxidase(PNPO) gene mutations in 2 patients with PNPO deficiency.Methods The identical twin brothers were diagnosed at the Department of Pediatrics of Peking University First Hospital in February 2016.The clinical presentations,course of treatment,blood biochemistry,metabolic screening,EEG,brain magnetic resonance imaging (MRI) and epilepsy-related genes detection (including PNPO gene) of them were analyzed.Results These 2 patients were born at 35 +5weeks gestation and had asphyxia after birth.The seizures started within the first day,which was uncontrolled by various antiepileptic drugs.EEG showed atypical hypsarrhythmia or multifocal epileptiform discharges.MRI showed nonspecific abnormality.Pyridoxine was used as monotherapy or combination with various antiepileptic drugs during the treatment.Seizures had ever been controlled by pyridoxine alone for up to 1 month.Antiepileptic drugs were withdrawn gradually under the circumstances of seizures persisting when they became 5 years old.During the past year,pyridoxine was used alone.They still had seizures at their age of 6 years and 4 months.Blood metabolic screening showed that the level of arginine,asparaginic acid and methionine decreased.Urinary metabolic screening showed vanillic acid elevating prominently in both patients,49.78 and 36.60 times beyond normal,respectively.Genetic analysis showed compound heterozygous variants ofPNPO gene in both patients:c.445_448del (p.P150RfsX27) and c.481C ＞T (p.R161C).These 2variants were not reported previously.After definite diagnosis was made,pyridoxine was replaced by pyridoxal-5'-phosphate (PLP) immediately.Seizures increased slightly at the initial treating with PLP,then reduced gradually and were controlled eventually.Psychomotor development was severely delayed in 2 patients.Conclusions Infantile onset intractable seizures in these 2 patients responded to pyridoxine.The results of blood and urinary metabolic screening suggest the possibility of PNPO deficiency.This is the first time to report patients with PNPO deficiency diagnosed by PNPO gene mutations in China.Seizures could be controlled by PLP monotherapy eventually.

Objective To explore the therapeutic effects and adverse reaction of high-dose Diazepam (DZP) in patients with electrical status epilepticus during sleep (ESES).Methods Nine patients in the Outpatient of the Department of Pediatrics,Peking University First Hospital from October 2016 to May 2017 with ESES were treated with high-dose DZP.Oral DZP was administered in a dose of 0.75-1.00 mg/kg(maximum:40 mg) during the first night followed by 0.5 mg/(kg · d) (maximum:20 mg) from the second night for 1-3 months and tapered over next 1-3 months.The seizures,electroencephalogram (EEG) changes and adverse reactions were observed before and after DZP treatment.Results Six of 9 patients were male and 3 were female.The age of onset was ranged from 1 year and 6 months to 10 years.Benign childhood epilepsy with central temporal spike was diagnosed in 5 cases,epileptic encephalopathy with continuous spike-and-wave during sleep in 1 case,and ESES related epilepsy in 3 cases.Age of onset DZP treatment ranged from 4 years and 4 months to 12 years,and the duration of DZP treatment was ranged from 1 to 5 months (1 case only for the first night).The follow-up interval was 6-12 months.The efficiency of DZP on seizures:intent effective in 5 patients,effective in 2 patients and ineffective in 2 patients,and the effective rate was 78％ (7/9 cases).The efficiency of DZP on EEG (1 month after DZP treatment):intent effective in 2 patients (EEG normalized),effect in 3 patients and no effect in 2 patients,and the effective rate was 71％ (5/7 cases),while 2 patients did not receive EEG examination.Four of 7 patients (57％) with intent effect and effective of DZP on seizures had seizures relapse during drug reduction and after drug withdrawal,and the EEG deteriorated simultaneously.Adverse reactions of DZP included 3 patients (33％) with adverse reactions,bed-wetting in 2 patients and snoring on the first night in 1 patient who withdrew DZP later.Conclusions The high-dose of DZP has a certain effect on seizures control and ESES suppression in patients with ESES,but also has a certain recurrence rate.The adverse reactions are mild and self-limiting.High-dose DZP treatment could be a choice for refractory patients with ESES to alleviate disease.

Objective: To investigate the electroclinical findings in epilepsy children with epileptic negative myoclonus (ENM) restricted to the lower limb as the first seizure type. Methods: Each retrieved electroencephalogram record performed between March 2011 and March 2018 at the Department of Pediatrics of Peking University First Hospital was searched with "midline" . There were 302 records of 175 patients with "benign" or "functional" midline spikes. A retrospective review of each patient′s hospital record was performed. Thirteen patients had ENM restricted to the lower limb as the first seizure type. The clinical and electroencephalogram characteristics of them were analyzed. Results: Thirteen patients manifested ENM restricted to the lower limb as the first seizure type, diagnosed as benign childhood focal epilepsy with vertex spikes (BEVS). Six patients had ENM as the first and only seizure type during the short-time follow-up. Among them, there were 1 male and 5 females. The age at seizure onset was (2.5±0.7) years. One of them had electrical status epilepticus during sleep (ESES) identified on electroencephalogram at theage of 4 years and 8 months. The last follow-up age was (3.8±1.5) years. The remaining 7 patients developed nocturnal focal motor seizures. Among them, there were 4 males and 3 females. The age at seizure onset was (3.5±0.7) years. Two of them were diagnosed as BEVS evolving into benign childhood epilepsy with centrotemporal spikes (BECTS) and 5 were diagnosed as BEVS concurring with BECTS. The age at focal seizures was (4.1±0.6) years. The interval ranged from 1 month to 1 years. Six of 7 patients had electrical ESES with the age of (5.2±1.0) years. All had developmental regression, further diagnosed as atypical benign partial epilepsy (ABPE). The median age at last follow-up was 5.9 years. Five of 13 patients had repeated electroencephalogram records at our apartment, showing that epileptiform discharges in midline regions were significantly reduced either in frequency or amplitude with the improvement of ENM restricted to the lower limb and that independent epileptiform discharges in Rolandic regions from midline regions were noticed with the onset of nocturnal focal seizures. Conclusions ENM restricted to the lower limb has a close association with vertex (midline) epileptiform discharges. ENM restricted to the lower limb as the first seizure type is a peculiar phenomenon of BEVS. Some patients could evolve into BECTS or overlap with BECTS, and further into ABPE. The age of seizure onset in BEVS with ENM restricted to the lower limb as the first symptom is a little earlier than in BECTS. Ignorance of the close association between midline spikes and ENM restricted to the lower limb may lead to misdiagnosis of these patients.

Objective:To summarize the clinical phenotype and gene mutation characteristics of male patients with epilepsy caused by mosaic PCDH19 mutation. Methods:The clinical data of 3 male patients with epilepsy caused by mosaic PCDH19 mutation were analyzed.Microdroplet digital polymerase chain reaction (mDDPCR) was used for the detection of mosaicism in the three probands and their family members.Relevant literatures were reviewed. Results:The seizure onset age were 5 months, 9 months and 6 months of life respectively.Focal seizures occurred in 2 cases and multiple seizure types occurred in 1 case.Three patients presented with clusters of seizures.Fever sensitivity was observed in 2 cases out of the 3 cases.Two patients had intellectual disability and 1 patient had autistic manifestation.The clinical phenotype in 2 patient fulfilled the diagnosis of Dravet syndrome. PCDH19 mosaic mutations c. 317T>A(p.M106K), c.158dupT(p.D54Gfs*35) and c. 1639G>C(p.A547P) were detected respectively, and were identified as de novo after parental validation.Mutant allele fractions (MAF) in the blood samples were identified as 81.18%, 37.08%, 77.64%, respectively.The MAF of multiple tissues in 1 patient varied from 78.67% to 98.46%.Review of literature revealed that a total of 11 cases with mosaic PCDH19 mutation were reported.Among them, seizure onset occurred between 5 and 31 months of age.Focal seizures occurred in 9 cases, 3 cases of the 9 cases had only focal seizures.Generalized tonic clonic seizures occurred in 4 cases.Two or more seizures were observed in 6 cases.Clustering of seizures was found in all patient and sensitivity to fever was observed in 9 patients.Seven patients had mild to severe intellectual disability and 5 patients had autistic features. Conclusions:The clinical phenotypes of male patients with epilepsy caused by PCDH19 mosaic mutation are characterized by clustering of seizures, sensitivity to fever, focal seizures in most cases, varied degree of intellectual disability and autistic features in partial.