Objective To analyze the clinical,muscle pathological features and molecular mutations in the 2 Chinese Han siblings with limb-girdle muscular dystrophy(LGMD) and conclude the phenotype/genotype correlations.Methods Clinical and muscle pathological data were collected.Genomic DNA of the two siblings and their parents were extracted using standard procedures from the peripheral leukocytes.A custom of targeted gene panel including 61 neuromuscular genes were designed by using the Agilent Sureselect Target Enrichment Kit.Targeted next generation sequencing(NGS) was performed in the proband,and CAPN3 gene mutation was verified with Sanger sequencing in the two siblings and their parents.The dbSNP138 and http://www.dmd.nl were searched to determine the disease-causing mutations.Results The proband slowly showed muscle weakness profoundly with pelvic muscles,developed difficulty in squatting and standing and climbing stairs.She had a tight Achilles tendon,high CK level (1 908-9 241 IU/L),without winging scapula and hypertrophy calf.The affected brother was only diagnosed hyper CKemia.By using the targeted NGS,the two siblings possessed the same two compound heterozygous mutations(c.717delT and c.2243G ＞ A) in CAPN3 gene.The two mutations both were verified by Sanger sequencing and had been reported before.Conclusions LGMD is clinically and genetically heterogeneous,and targeted NGS is powerful in defining the causal mutation of LGMD and helpful in investigating the exact genotype/phenotype analysis.

Bladder cancer is a common malignant tumor in urinary system. The life quality of patients reduces obviously after radical resection of bladder. Comprehensive treatment including radiotherapy and chem-otherapy after bladder preservation surgery plays an important role for the prevention of postoperative recur-rence,preservation the function of bladder,and improving the life quality of patients. Image-guided radiothera-py can reduce the setup error and inner boundary caused by the movement of organs,and can alleviate the side reaction of radiation,and it also can provide basis of expanding boundary of planning target volume for the blad-der cancer patients.

Objective:To compare the effects ofpropofol and sevoflurane on the plasma thromboxane B2 (TXB2),endothelin-1 (ET-1) and D-dimer (D-D) levels of patients underwent posterior retroperitoneal laparoscopic surgery.Methods:84 cases of patients underwent post retroperitoneal laparoscopic surgery in our hospital from May 2015 to December 2016 were selected as research objectives and randomly divided into two groups with 42 cases in each group.The same anesthesia induction were provided for two groups,the observation group was given 2％～3％ sevoflurane for continuous inhalation,while the control group was given 4～12 mg/(kg·h) of propofol for continuous injection by pump.Both groups received remifentanil 10 μg/ (kg ·h) target-controlled infusion simultaneously.The levels of plasma TXB2,ET-1 and D-D in the two groups were measured after anesthesia induction (T0),at 0.5 h (T1),1 h (T2),1.5 h (T3) after pneumoperitoneum.Meanwhile,the anesthetic effects and adverse reactions were compared between two groups.Results:The time of consciousness disappearence,time of tracheal intubation,spontaneous breathing recovery time,eye opening time,verbal response time,orientation recovery time and extubation time of observation group were significantly shorter than those of the control group (P＜0.01).No significant difference was found in the occurrence of adverse reactions between two groups (P＞0.05).The plasma TXB2,ET-1 and D-D levels of both groups were gradually increased at T1,T2 and T3,and all were significantly higher than that at T0 (P＜0.01).The plsma TXB2,ET-1 and D-D levels at T1,T2 and T3 of observation group were significantly lower than those of the control group at same time (P＜0.01).Conclusion:Posterior laparoscopic surgery could cause different degrees of hypercoagulability of blood.Compared with propofol,sevoflurane could effectively inhibit the release of TXB2,ET-1 and D-D in anesthesia after retroperitoneal laparoscopic anesthesia,and play a better role of anticoagulation.

Objective To investigate the acute toxicity of intravenous isoflurane in Beagles.Methods Six healthy adult Beagles of both sexes aged 6-8 months weighing 6-8 kg were used in this study.Isoflurane injectio (120 mg/ml) in 30% hpid emulsion was injected intravenously. Femoral artery was cannulated for direct BP monitoring.ECG was continuously monitored.The maximal tolerance dose (MTD) and approximate lethal dose (ALD) were determined by up-and-down technique. The initial dose was 3.0 ml/kg. The dose was decreased/increased by 0.3 ml/kg if the previous animal died/survived.The survived dogs were observed for 2 weeks.Autopsy and histopathological examination were performed on all dead Beagles.Results The ALD and MTD of intravenous isoflurane were 252 and 216 mg/kg. Autopsy and histopathological examination did not show any abnormality.Conclusion Cardiopulmonary depression is the main manifestation of the acute toxicity of intravenous isoflurane in Beagles.

Objective To investgate the effects of dexmedetomidine combined with small-dose of sufentanil on the hemodynamics during anesthesia induction in patients undergoing off-pump coronary artery bypass grafting.Methods Seventy-five ASA Ⅱ or Ⅲ and NYHA Ⅱ or Ⅲ patients aged 46-72 yr,weighing 59-86 kg,ejection fraction ≥45％,undergoing off-pump coronary artery bypass grafting,were randomly divided into 3 groups ( n =25):dexmedetomidine combined with small-dose of sufentanil group (group DS),small-dose of sufentanil group (group S1 ) and large-dose of sufentanil group (group S2 ).In group DS,dexmedetomidine 0.8 μg/kg (diluted with normal saline to 15 ml) was injected for 15 min at a rate of60 ml/h,while the same volume of normal saline were given in groups S1 and S2.Anesthesia was induced with midasolam 0.08 mg/kg and pipecuronium 0.12 mg/kg.After administration of the total dose of 1/3 midazolam and 1/8 pipecuronium,sufentanil 0.5,0.5 and 0.8μg/kg (diluted with narmalsaline to 10 ml) were injected in groups DS,S1 and S2 respectively.Then the rest of midazolam was injected.When BIS value ≤ 75,the rest of pipecuronium was injected.When BIS value ≤ 55,the patients were tracheal intubated and mechanically ventilated.PETCO2 was maintained at 30-35 mm Hg.The adverse ardiovascular events (hypertension,hypotension,tachycardia and bradycardia) and drugs intervention were recorded during anesthesia induction.Results Compared with group S2,the incidence of hypertension and tachycardia was significantly increased in group S1,the incidence of hypotension decreased in groups S1 and DS,the incidence of drug intervention decreased in group DS (P ＜ 0.05).Compared with group S1,the incidence of hypertension,hypotension and tachycardia was significantly decreased,the incidence of bradycardia increased,theincidence of drug intervention decreased in group DS (P ＜ 0.05).Conclusion Dexmedetomidine (0.8 μg/kg) combined with small-dose of sufentanil (0.5 μg/kg) is beneficial for keep the stable of hemodynamics during anesthesia induction in patients undergoing off-pump coronary artery bypass grafting.

[ ABSTRACT] AIM:To study the expression of WNT5B in the breast cancer and further to discuss the correlation between WNT5B and clinicopathologic characteristics of breast cancer.METHODS:The expression of WNT5B at mRNA and protein levels was measured by real-time PCR and Western blot in 67 cases of breast cancer and the tissue adjacent to carcinoma.In addition, the immunohistochemical method was used to detect the expression of WNT5B in the breast cancer and the tissue adjacent to carcinoma.The relationships between WNT5B expression and clinicopathologic indexes were also analyzed.RESULTS:The expression of WNT5B in the breast cancer was obviously lower than that in the tissue adjacent to carcinoma (P<0.05).The expression of WNT5B at mRNA and protein levels in 67 samples of breast cancer was in va-rious degrees.The expression of WNT5B in T≤20 mm group of human breast cancer was obviously higher than that in T>20 mm group (P<0.05).The expression of WNT5B had no obvious correlation with axillary lymph node metastasis, histo-logical grade and immunohistochemical indexes of ER, PR, c-ErBb-2, p53 and Ki67 ( P>0.05) in the breast cancer. CONCLUSION:The expression of WNT5B decreases obviously in breast cancer.The expression of WNT5B is related to primary tumor size, which provides new ideas for the diagnosis and treatment of breast cancer, suggesting that WNT5B may be a new molecular marker for prognosis of breast cancer.

Objective To evaluate the effects of dexmedetomidine on hemodynamics during induction of anesthesia in the patients with atrial fibrillation with rapid ventricular rate undergoing noncardiac surgery.Methods Fifty patients with rheumatic valvular heart disease complicated with atrial fibrillation,aged 45-64 yr,weighing 50-75 kg,with ventricular rate ≥ 90 bpm,of ASA physical status Ⅱ or Ⅲll (NYHA Ⅱ or Ⅲ),scheduled for elective surgery,were randomly divided into 2 groups (n =25 each) using a random number table:control group (group C) and dexmedetomidine group (group D).Dexmedetomidine 0.6 μg/kg was infused intravenously at 10 min prior to induction of anesthesia in group D.Anesthesia was induced with iv midazolam 0.06 mg/kg,sufentanil 0.6 μg/kg,and vecuronium 0.12 mg/kg.Tracheal intubation was performed when the BIS value≤≤ 55After admission to operating room (T0,baseline),immediately after the end of dexmedetomidine infusion (T1),immediately before intubation (T2),and at 1,3 and 5 min after intubation (T3-5),SP,DP,MAP and HR were recorded.The adverse cardiovascular events were recorded starting from induction of anesthesia to 5 min after intubation.Results Compared with the baseline value at T0,HR was significantly decreased at T2,5,while increased at T3,4 in group C,and HR was decreased at T1-5 in group D; SP,DP and MAP were decreased at T2,5,while increased at T3 in group C,and no significant changes were found in the indices mentioned above in group D.Compared with group C,the incidence of hypotension,hypertension and tachycardia was significantly decreased,and no significant changes were found in theincidence of bradycardia in group D.Conclusion Dexmedetomidine 0.6 μg/kg infused intravously is helpful in maintaining the hemadynamics stable during induction of anesthesia in the patients with atrial fibrillation with rapid ventricular rate underging noncardiac surgery.

PCDH19 gene related epilepsy is an unusual X - linked disease that females and mosaic males are affected,while hemizygous males are not. Recently,the number of reports about PCDH19 gene related epilepsy is in-creasing,and PCDH19 gene has become one of the most important epilepsy genes. Now,the structure and function of PCDH19 gene and protein,the inheritance and pathogenesis,clinical features,treatment and genotype/ phenotype asso-ciated with PCDH19 gene related epilepsy,were reviewed.

Objective: To investigate the spectrum of mutations in families with benign familial neonatal-infantile epilepsy (BFNIE) . Methods: Clinical data and peripheral blood DNA samples of all BFNIE probands and their family members were collected from Peking University First Hospital between December 2012 and April 2016. Clinical phenotypes of affected members were analyzed. Genomic DNA was extracted from peripheral blood samples with standard protoco1. Mutations in PRRT2 were screened using Sanger sequencing. For families that PRRT2 mutations were not detected by Sanger sequencing, candidate gene mutations were further screened by next-generation sequencing for epilepsy. Results: A total of 7 families were collected. Of the 30 affected members, 15 were male and 15 were female. The age of epilepsy onset was from 2 days to 6 months. Genetic testing led to the identification of gene mutations in all families. One family had the PRRT2 hotspot mutation (c.649dupC). Three families had missense SCN2A mutations (c.2674G>A/p.V892I, c.2872A>G/p.M958V, and c.2627A>G/p.N876S) . Both c.2872A>G/p.M958V and c.2627A>G/p.N876S were novel SCN2A mutations. Three families had KCNQ2 mutations. Two of them had missense mutations (c.958G>A/p.V320I and c.998G>A/p.R333Q) . The KCNQ2 mutation c.958G>A/p.V320I was novel. One family had a gene deletion of KCNQ2, which also extended to the adjacent gene, CHRNA4; and the deletion involved all the exons of KCNQ2 and CHRNA4. Conclusions Mutations in KCNQ2, SCN2A, and PRRT2 are genetic causes of BFNIE in Chinese families. The detection rate for gene mutations is high in BFNIE families. KCNQ2 and SCN2A mutations are common in BFNIE families. SCN2A mutations (c.2872A>G/p.M958V and c.2627A>G/p.N876S) and KCNQ2 mutation (c.958G>A/p.V320I) are novel mutations.

Objective: To summarize the phenotype of epileptic children with SCN2A mutations. Methods: Epileptic patients who were treated in the Pediatric Department of Peking University First Hospital from September 2006 to October 2017 and detected with SCN2A mutations by targeted next-generation sequencing were enrolled. Clinical manifestations of all patients were analyzed retrospectively. Results: A total of 21 patients (16 boys and 5 girls) with SCN2A mutations were collected. Twenty-one SCN2A mutations were identified. Ten patients had mutations inherited from one of their parents and 11 patients had de novo mutations. The age of epilepsy onset was from 2 days to 2 years and 6 months: six patients with seizure onset in neonates (29%) , six patients with seizure onset between 1 month and 3 months of age (29%), three patients with seizure onset between 4 months and 6 months of age, two patients with seizure onset between 7 months and one year of age, and four patients with seizure onset beyond one year of age. Multiple seizure types were observed. The focal seizure was the most common seizure type which was observed in 18 patients (86%) . Spasm seizure was observed in 6 patients (29%) . Other seizure types were rare. In 19 patients, seizures manifested in clusters (90%) . In 3 patients, seizures manifested fever-sensitive. Nine of ten patients with inherited SCN2A mutations had normal development. However, all patients with de novo SCN2A mutations had mild or severer development delay. In 21 patients with SCN2A mutations, five were diagnosed with benign familial infantile epilepsy, 3 with benign familial neonatal-infantile epilepsy, 3 with Ohtahara syndrome, 3 with West syndrome, 2 with encephalopathy with early infantile onset epilepsy, one with febrile seizures plus, one with Dravet syndrome, one with encephalopathy with childhood-onset epilepsy, one with autism with epilepsy and one with intellectual disability with epilepsy. Conclusions The clinical features of patients with SCN2A mutations include that main seizure onset is the neonate and early infancy, and the main seizure type is the focal seizure, manifested in clusters. The large spectrum of SCN2A-related epilepsy, which not only includes epilepsy with a comparatively favorable prognosis, but also epileptic encephalopathy. De novo mutations often lead to severe phenotype with development delay.