Objective To find out some ways to solve the problems which are dealing with the clinical application of intrafix.Methods From February to May 2013,324 patients with back blood using intrafix in emergency department were randomly assigned to the experimental group A and the control group A,with 162 patients in each group.The experimental group A used the method of pressing pipe while pulling out the needles when transfusion was over,while the control group A used traditional ways.206 patients who occurred bubbles in the intrafix among the above two groups were divided into the experimental group B and the control group B.The experimental group B got rid of the bubbles using top-down method,the control group B used traditional ways.The effect of withdrawing the needles and exhaustion of bubbles were compared.The least flushing time of remaining liquid using transfusion pipe flushing method was calculated.Results The experimental group A and the experimental group B were better in withdrawing the needles and exhaustion of bubbles compared with those of the control group A and the control group B.Using the method of drop coefficient to calculate the least flushing time can ensure the drug to be transfused into the body thoroughly.Conclusions When transfusion is over,pressing intrafix pipe will have a better result than the traditional way when pulling out the pipe.Top-down method can clearly settle the problem of intrafix pipe,which can not get rid of bubble via traditional way.Flushing pipe and press pipe method can apparently decrease drug waste.

Objective:To analyze the effect of simvastatin combined with tanshinone in the treatment of patients with acute cere-bral infarction. Methods:The patients with acute cerebral infarction were randomly divided into the treatment group (n=50) and the control group (n=50). The control group was given tanshinone treatment, the treatment group was given simvastatin treatment addi-tionally, and the treatment course was 14 d. The NIHSS ( nerve function defect degree) , ADL ( daily life activity) and the treatment effect before and after the treatment were comprehensively evaluated and compared between the two groups. Results: Compared with those before the treatment, the NIHSS and ADL scores were significantly improved in the two groups after the treatment, and the differ-ences were statistically significant (P<0. 05), and the improvement in the treatment group was significantly better than that in the control group with statistical significance (P<0. 05). The total effective rate of the observation group was 98%, which was higher than that (70%) in the control group (P<0. 05). Conclusion: Simvastatin combined with tanshinone in the treatment of acute cerebral infarction shows obvious therapeutic effect, which can obviously improve neurologic deficits and daily life activity, and is worthy of fur-ther clinical application.

Objective To study the relationship between insulin resistance and carotid atherosclerosis in elderly patients with type 2 diabetes.Methods A total of 63 elderly patients with type 2 diabetes were divided into 2 groups:group A (HbA1c≤7％,n=30),group B (HbA1c＞7％,n=33); and 30 healthy people were as controls.Fasting blood sugar(FBS),fasting insulin (FINS),glycated hemoglobin (HbA1c) and carotid intima-media thickness (IMT) were measured,atherosclerotic plaques were counted,and insulin resistance was calculated by homeostatic model assessment (HOMA-IR).Results The IMT,number of atherosclerotic plaques and incidence of carotid atherosclerosis were all higher in the two diabetic groups than in controls (P＜0.05).The levels of FINS,HbA1c and HOMA-IR were all higher in group A and group B than in controls (P＜0.05 and P＜0.01),which had a significant difference between group A and group B [(9.7± 2.1)mU/L vs.(13.6±2.0) mU/L; (6.5±0.4)％ vs.(8.2±0.6)％; (3.5±0.4) vs.(6.1±0.5); all P＜0.05].Pearson correlation analysis showed that IMT was positively correlated with FBS and HOMA-IR in group A and group B (r=0.62 and r=0.46,respectively,P＜0.05).Conclusions There is a positive correlation between insulin resistance and carotid atherosclerosis in elderly patients with type 2 diabetes.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity. The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD). Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development, differentiation, survival and function. Meanwhile, GDNF mutations are a major cause of HD. In this study, BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3, and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM). Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5, VIP and nNOS. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs. The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM. This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders, such as HD.

Background Leber hereditary optic neuropathy (LHON) is a maternally inherited disease caused by mitochondrial DNA (mtDNA) mutation with the common mutation sites of m.3460 G＞A,m.11778 G＞A and m.14484 T＞C,and other mutation sites are rare.Understanding the mutation type of mtDNA in LHON patients has an important clinical significance.Objective This study was to analyze the clinical features of LHON and detect the mitochondrial mutation.Methods Twelve unrelated Chinese patients who was diagnosed as LHON were included in Peking University People's Hospital from 2010 to 2014.The visual acuity,perimetry,ocular segment,visual evoked potential,fundus were binocularly examined.The peripheral blood of 4 ml was collected from each patient and mtDNA was amplified and sequenced by using PCR.Three common genetic mutation sites for LHON and other mutation sites were determined and analyzed.This study protocol was approved by Ethic Committee of Peking University People's Hospital and complied with Helsinki Declaration.Written informed consent was obtained from each patient prior to any medical examination.Results Of the 12 patients,11 were male and 1 was female.The visual acuity of both eyes reduced simultaneously in 7 patients,and the visual acuity of left eye and the right eye first reduced in 3 patients and 1 patient,respectively.There was no significant correlation in the visual impairment between the left and right eyes (P＞0.05).In the near vision of the patients,J7 was invisible in 18 eyes,and J7 were obtained in 3 eyes,J6 were obtained in 2 eyes and J2 was obtained in 1 eye.In the distant vision of the patients,hand movement was obtained in 1 eye,light perception was obtained in 1 eye,0.01-0.1 were obtained in 18 eyes and 0.12-0.3 were obtained in 2 eyes.The visual field defect of nasal lateral was found in 7 eyes,visual field defect of temporal lateral was found in 3 eyes and the visual field defect of central was found in 8 eyes.mtDNA sequencing revealed that m.3460 G＞A mutation was seen in 3 patients,m.11778 G＞A mutation was seen in 5 patients and m.14484 T＞C mutation was seen in 2 patients.In addition,other 2 mutations were found in 2 patients,which were m.3497 C＞T and m.10663 T＞C mutations at the MT-ND1 and MT-ND4L genes,respectively.Conclusions LHON is more common in male.Visual impairment shows the varying degrees between both eyes of patients and appears to be severe in near vision.Central visual field defect is common in LHON patients.This study detects m.3497 C＞T and m.10663 T＞C mutation in Chinese LHON patients.

Objective To set up a new method,which is sensitive,low cost,rapid and suitable for clinical application for FTO gene rs9930506 variant genotyping basing on high resolution melting (HRM) platform,and to preliminarily put into practice in susceptibility analysis for metabolic syndrome (MS) in Beijing.Methods Unlabelled probe with C3-spacer block specific for rs9930506 variant has been designed according to the Refseq from GenBank.With LC-Green plus dye pre-mixed,we scanned the signal for the genotype analysis after PCR amplification and HRM reaction.Restriction fragment length polymorphism (RFLP) and PCR-sequencing methods were designed as 2 control genotyping methods for the evaluation of accuracy and convenience.Afterwards,the HRM-based method was put into practice in metabolic syndrome patients (n =500) and control groups (n =500) for rs9930506 genotyping,and primarily study the association between rs9930506 and MS.Results All the 3 methods could genotype rs9930506 appropriately,although the 2 control methods seemed to be a little time-inefficient.The call rate of HRM-method was 100％ and sampling accuracy reached 99.3％ according to sequencing results.In the MS group,AA,AG and GG genotypes were found in 290,185 and 25 cases,respectively.And in the control group,those were found in 344,138 and 18 cases.No genotype distribution difference was detected between control group and HapMap-CHB data (P =0.520 ).The genotype distributions were all in Hardy-Weinberg equilibrium in each group.AA genotype of rs9930506 seemed to reduce the risk for MS( OR =0.626,95％CI =0.483-0.812).Conclusions The AA genotype of rs9930506 variant in FTO might be a protective factor for MS in Beijing population.The susceptibility related genotyping in clinical samples could be more rapid,precise and inexpensive with the development of HRM in genotyping.

ObjectiveTo evaluate the therapeutic efficacy of low-dosage methyltestosterone or andriol in men with senile osteoporosis. MethodsA total of 134 male patients with senile osteoporosis and the decreased serum level of free testosterone were divided into three groups. 45 patients were treated with low-dosage methyhestosterone(100 mg, once a day, sublingual) and 46 patients were treated with low-dosage andriol (40 mg, once a day, orally), while 43 patients were treated with placebo. The duration of treatment in each group was 1 year. The bone density, blood and urine biochemical indexes related to bone metaholites,the quality of life indexes, ultrasonography for prostate,serum prostate specific antigen,blood routine, urine routine, hepatic and renal function were detected before and after the treatment. ResultsBoth low-dosage methyltestosterone and low-dosage andriol could prevent the decrease of bone mineral density and improve patients' general health, role-emotional function and vitality (all P＜0.05). The difference values of femoral neck bone mineral density before and after treatment with low-dosage andriol and low-dosage methyltestosterone were (0.14+0.18)g/cm2 and (0.12±0.09)g/cm2 , respectively(P＜0.05). Low-dosage andriol hadstronger effects in increasing the level of estradiol (32.5±14.2 )ng/L than low-dosage methyltestosterone(19.3±9.2)ng/L(P＜0.05) and showed more notable effects in improving the physical functioning and role-physical function than low-dosage methyhestosterone. The use of the two androgenic hormones at low dosage showed safety. ConclusionsBoth low-dosage methyltestosterone and low-dosage andriol can be used to treat senile osteoporosis in men and to improve life quality. Both of them are effective and safe therapeutic choices.

AIM:To study the effect of Panax quinquefolium saponin (PQS) on cardiomyocyte apoptosis in-duced by thapsigargin ( TG) .METHODS:Primary cultured cardiomyocytes from neonatal SD rats were divided into con-trol group, TG group, PQS (40 mg/L, 80 mg/L and 160 mg/L) +TG group, si-PERK+TG group, and mock+TG group.The cells were treated with 1 μmol/L TG for 24 h to induce apoptosis.The PERK gene in the cardiomyocytes was knocked down by RNAi.The cell viability was detected by CCK-8 assay.Apoptosis was analyzed by flow cytometry.Wes-tern blotting was used to determine the expression of ERS molecules GRP78, CRT, ATF4 and CHOP, anti-apoptosis pro-tein Bcl-2 and pro-apoptosis protein Bax.RESULTS: Compared with control group, TG significantly and the apoptosis, reduced the cell viability (P<0.05), increased the phosphorylation of PERK and eIF2α, increased the expression of GRP78, CRT, ATF4, CHOP and pro-apoptosis protein Bax, and decreased the expression of anti-apoptosis protein Bcl-2 (P<0.05).Compared with TG group, PQS treatment (160 mg/L) significantly reduced the apoptosis and increased the cell viability (P<0.05).All the 3 different concentrations of PQS significantly increased the expression of anti-apoptosis protein Bcl-2 and reduced the expression of pro-apoptosis protein Bax (P<0.05) in a dose-dependent manner.PQS pre-treatment and knockdown of PERK both reduced the protein levels of GRP78, CRT, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP and pro-apoptosis protein Bax, and increased the expression of anti-apoptosis protein Bcl-2 (P<0.05). CONCLUSION:PQS at concentration of 160 mg/L attenuated cardiomyocyte apoptosis induced by TG.PQS had the simi-lar effect as PERK knockdown on cardiomyocyte apoptosis.The mechanism may be associated with inhibiting PERK-eIF2α-ATF4-CHOP pathway of ERS-related apoptosis.

Base excision repair (BER) is an important pathway for DNA oxidative damage repair,and mutations in this pathway cause gene instability and increase cancer risk.A large number of studies have shown that abnormal expression of key proteins in the BER pathway in a variety of tumors,such as DNA glycosylase,apurinic/apyrimidinic endonuclease 1,DNA polymerase and so on.Single nucleotide polymorphisms of these proteins affect the repair activity of tumor cells,and are expected to be important indicators for the diagnosis,treatment and evaluation of prognosis.

Objective To investigate the effect of methylene blue (MB) on oxygen metabolism in patients with septic shock. MethodsForty ASA Ⅱ or Ⅲ patients with septic shock aged 38-64 yr weighing 48-75 kg undergoing emergency surgery were randomly divided into 2 groups ( n = 20 each): group Ⅰ norepinephrine (group NE) and group Ⅱ MB. The patients were unpremedicated. Anesthesia was induced with midazolam, etomidate,sufentanil and vecuronium and maintained with inhalation of 0.5％-1.5％ sevoflurane and intermittent iv boluses of sufentanil and vecuronium. The patients were mechanically ventilated after tracheal intubation. PETCO2 was maintained at 35-45 mm Hg. During operation MB was infused at 0.5-1.0 mg·kg-1·h-1 in group MB and NE at 0.5-2.0 μg· kg-1 · min-1 in group NE respectively to maintain hemodynamic stability. Radial artery was cannulated and connected with Vigileo cardiac output monitor. Right internal jugular vein was cannulated for CVP monitoring.HR, SvO2, MAP, CVP, SV and CI were continuously monitored. Arterial and central venous blood samples were collected simultaneously before induction of anesthesia (T0, baseline), immediately before (T1) and at 30, 60 and 90 min after skin incision (T2-4) and at the end of operation (T5) . Blood gas analysis was performed. O2 consumption index (VO2I), O2 delivery index (DO2I) and O2 extraction rate (ERO2) were calculated. Blood lactate concentration was measured. Results MAP, HR, CVP, SVRI, DO2I, VO2I and ERO2 were significantly higher,while CI and blood lactate concentration lower during operation (T2-5) in group MB than in group NE. MAP, HR,CVP, SVRI, VO2I, DO2I, and ERO2 were significantly higher, while CI and blood lactate concentration were lower during operation (T2-5) as compared with the baseline values at T0 in group MB. In group NE there were no significant change in MAP, HR, CVP and DO2I during operation (T2-5 ) as compared with the baseline at T0. ConclusionIntravenous infusion of MB at 0.5-1.0 mg·kg-1·h-1 during operation may improve hemodynamics and oxygen metabolism in patients with septic shock.