Objective The purpose of this study is to investigate the molecular mechanisms of p.C310R(c.T928C) and p.E396V(c.A1187T) lipoprotein lipase(LPL) gene mutations in vitro, which may help to construct the spectrum of LPL gene mutations and phenotype. It also can provide accurate early diagnosis for high-risk population of familial hypertriglyceridemia and provide the basis for the development of gene targeted therapy. Methods Genomic DNA was extracted from proband′s family members′ peripheral blood cells and screened by whole-exome sequencing to verify candidate gene variations. PCR products were afterwards directly sequenced again to confirm corresponding LPL variants. At the cellular level, lentiviruses containing LPL mutations were constructed and then transfected into COS-1 cells. Functional significance of the mutants was corroborated by analyzing LPL activity and mass in the cell medium and lysates via ELISA and enzyme-fluorescent method. mRNA was assayed by RT-PCR to confirm the effect on gene transcription. Results DNA sequence analysis revealed that the proband was a heterozygote for a novel c.T928C mutation in exon 6 of LPL gene, while his nephew was a compound heterozygote for the c.T928C mutation in exon 6 and a novel c.A1187T mutation in exon 8. In vitro studies, these two mutations can cause decreased activity and mass of extracellular LPL(P<0.05). Moreover, further investigation indicated that LPL C310R mutation tremendously affected post-transcriptional modification of LPL gene, whereas LPL E396V mutation dampened intracellular LPL trafficking. Conclusion Both the mutations are pathogenic by reducing the activity and mass of LPL in the plasma, which affected normal metabolism of triglycerides.

Objective To study the expression of Ca-A/K channel-related molecules glutamate receptor 2 and 1(GluR2/1) in hippocampus tissues of neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods A total of 60 7-day-old Sprague-Dawley rats were randomly divided into sham operation group and HIBD group. Hippocampal tissues were obtained at 0 h, 1 h, 6 h, 24 h, 48 h and 72 h after HIBD. The expression of GluR2, GluR1 and autophagy marker protein Beclin-1, LC3 were detected by Western blot assay. Results Edema and focal softening and necrosis were observed 6 h after HIBD in the brains of neonatal rats. Compared with Con group, at each time point, the expression levels of GluR2 were lower while the levels of GluR1, Beclin-1 and LC3 were higher significantly in HIBD group (P<0.05). The protein levels of LC3, Beclin-1, GluR1 and GluR2 in hippocampus tissues of HIBD group were significantly different among different time points after the estab-lishment of HIBD model (F=10.65~701.14, P<0.01). The protein level of GluR2 was decreased from 1 h to 24 h after HIBD and reached the lowest level at 24 h. The levels of GluR1, Beclin-1 and LC3 were increased at 6 h, plateaued at 24 h and remained there until 48 h. The levels of these proteins returned back to the initial level at 72 h. Conclusions Ca-A/K channel-related mol-ecules GluR2 and GluR1 play important roles in the autophagic cell death of hippocampus tissues in neonatal rats with hypoxic-ischemic brain damage.

OBJECTIVETo evaluate the lumber plexus blockade as anesthesia technique for hip fracture repair in elderly patients.

METHODSWe retrospectively analyzed the peri-operative data of 87 hip fracture patients, aged 70 years or older, who underwent surgical repair at our hospital between 2003 and 2006. Patients were divided into three groups according the anesthesia techniques applied: general anesthesia (GA) group (n=21), epidural anesthesia (EA) group (n=37), and lumber plexus blockade (LPB) group (n=29).

RESULTSThe peri-operative data were comparable among three groups, except that intra-operative the dosage of fentanyl was significantly lower in LPB group compared with in GA group(P0.05), and the blood urea nitrogen 1 day after surgery was significantly increased in GA group(P0.05).

CONCLUSIONLumber plexus blockade combined with small-dose intravenous anesthesia is an optional anesthetic technique for elderly patients undergoing hip fracture repair.

Aged ; Aged, 80 and over ; Female ; Hip Fractures ; surgery ; Hip Joint ; surgery ; Humans ; Lumbosacral Plexus ; Male ; Nerve Block ; Retrospective Studies

OBJECTIVETo locate motor functional area of patients who undergone modified anatomical hemispherectomy in order to analysis the plasticity of upper limbs motor.

METHODSThe patients who undergone modified anatomical hemispherectomy were performed BOLD sequences, to locate functional cortical areas in their residual brain.

RESULTS6 patients have performed examination of BOLD sequences by 3.0-T MRI.5 of them obtained contralateral upper limb motor areas in their residual brain, and 3 of them obtained ipsilateral and contralateral upper limb motor area map in cortex. The ipsilateral upper limb motor areas in the M1, SMA and posterior parietal cortex.

CONCLUSIONSThe patients who undergone modified anatomical hemispherectomy is an excellent model to investigate mechanism of plasticity in the developing brain. Functional magnetic resonance (fMRI) provided fine spatial detail of brain responses, would describe the motor functional area of cortical maps. These patients exist ipsilateral motor areas in their residual mono hemisphere. The study indicated there maybe have somewhat extent of correlation between the surgical procedure and the outcome of neuroplasticity.

Cerebral Cortex ; physiopathology ; Epilepsy ; physiopathology ; surgery ; Female ; Hemispherectomy ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Neuronal Plasticity ; physiology ; Postoperative Period ; Upper Extremity ; innervation ; physiopathology

Objective To investigate the phenotypes and genotypes of a hereditary protein C(PC) deficiency pedigree.Methods Imrnunoassay(ELISA)was used for PC antigen and PS antigen; Immunoturbidimetry assay was used for measuring AT antigen;Chromogenic substrate assay was used for measuring the activity of PC,PS and AT in Sysmex 1500 automatic Blood Coagulation Analyzer.Polymerase chain reaction(PCR)for amplification of the fragment of each exon and side sequences of PC gene in 10 members of the 3 generations;Direct DNA sequencing was used to examine the mutation site.Results Among 10 members of the 3 generation pedigree,8 of them had a PC:Ag level of 1.06-1.92 mg/L(normal references 3.00-6.00 rag/L),the activity of PC was between 41% and 67%(normal references 70%- 140%),which was significantly lower than the normal references while the levels of PS:Ag,PS:A,AT:Ag and AT:A were all within normal range.DNA sequencing analysis showed that there was a G to T mutation in exon IX of the PC gene at 12 918 position in 8 members.This mutation resulted in the substitution of terminator TGA for TGG which encoding tryptophan at 372 amino acid.There was a polymorphism in 2 405C/ T,2 418A/G,2 583A/T in the promotor area.Conclusions This pedigree is a type I hereditary protein C deficiency.There is a G12 918T mutation in exon IX of PC gene.This mutation is reported for the first time and there is a polymorphism in 2 405C/T,2 418A/G,2 583A/T in the promotor area.

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

OBJECTIVETo investigate the influence of using fluid warming and forced-air warming system on patient core temperature, blood loss, blood transfusion, extubation time, and postoperative shivering.

METHODSForty ASA (American Society of Anesthesiologists' Physical Status) I-II patients, aged 21-69 years, scheduled for elective abdominal surgery under general anesthesia, were enrolled in the study. The patients were premedicated with intramuscular dolantin 50 mg and atropine 0.5 mg. Anesthesia was induced with midazolam 1 mg, fentanyl 50-100 microg and propofol 1.5-2.0 mg/kg. Tracheal intubation was facilitated with vecuronium 1mg and succinylcholine 1.5-2.0 mg/kg. The patients were mechanically ventilated and anesthesia was maintained with isoflurane 1.5-2.0%, 50% N2O in oxygen and intermittent iv boluses of fentanyl (total dose 5-6 microg/kg). Vecuronium was used for muscle relaxation during maintenance of anesthesia. The patients were randomly divided into 2 groups: control group (n = 20) and warming group (n = 20). In both groups, the patients were covered with surgery blanket. In the warming group, patients were additionally warmed with fluid warming device and forced-air warming system during the operation. The core temperature was recorded every 20 minutes during the operation, as well as the blood loss, blood transfusion, extubation time and postoperative shivering.

RESULTSThe core temperature at the end of the surgery was (36.4 +/- 0.4) degrees C in the warming group and (35.3 +/- 0.5) degrees C in the control group. The difference was statistically significant (t = 7.547, P < 0.001). There was no significant difference of blood loss and blood transfusion between two groups. The extubation time was significantly shorter in the warming group [(18 +/- 6) vs (26 +/- 10) min, t = -3.364, P = 0.002]. 6 patients shivered postoperatively in the control group and none in the warming group (chi2 = 7.059, P = 0.008).

CONCLUSIONFluid warming system and forced-air warming system can effectively maintain normothermia during the surgery and then help to reduce the extubation time and postoperative shivering.

Abdomen ; surgery ; Adult ; Aged ; Anesthesia, General ; Body Temperature ; physiology ; Female ; Humans ; Hypothermia ; prevention & control ; Intraoperative Care ; methods ; Intraoperative Complications ; prevention & control ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Shivering ; physiology

OBJECTIVETo evaluate the value of propofol target-controlled infusion combined with dribbled and nebulized lidocaine in tracheal intubation under spontaneous breathing.

METHODSTotally 40 elective surgery patients to accept tracheal intubation under unconsciousness and spontaneous breathing were randomly divided into 2 groups: 6-8 cm of endotracheal tube was inserted subglottic ally in the complete intubation group (n=20) while 3-4 cm was inserted temporarily in the partial intubation group (n=20).

RESULTSThe tracheal intubation was successfully completed under spontaneous breathing in all patients; meanwhile,the hemodynamic status was stable without any severe respiratory complications. Eleven patients suffered from moderate coughing response in the complete intubation group while no such response was noted in the partial intubation group (P<0.01).

CONCLUSIONSApplication of propofol target-controlled infusion combined with dribbled and nebulized lidocaine provides a good condition for tracheal intubation under unconsciousness and spontaneous breathing. The partial intubation can effectively prevent the occurrence of coughing response.

Adolescent ; Adult ; Aged ; Cough ; etiology ; prevention & control ; Female ; Humans ; Intubation, Intratracheal ; adverse effects ; methods ; Lidocaine ; administration & dosage ; Male ; Middle Aged ; Propofol ; administration & dosage ; Young Adult

BACKGROUNDIn the years around 1990, in Beijing Tiantan Hospital Affiliated to Capital Medical University many children with infantile hemiplegia and intractable epilepsy were treated with further modified anatomical hemispherectomy. We report the follow up of the first six cases. To make good use of these precious clinical data and make clear their neuropsychological state, we performed neuropsychological and neurophysiological measurements in these patients, who were at a median of 17.8 years after hemispherectomy.

METHODSOddball task was given to the patients and to a normal control group to collect the peak latency (PL) and peak amplitude (PA) of event-related potentials (ERPs)-P300. The P300 data of the two groups were analyzed and the P300 patterns of the six patients are presented. The baseline characteristics and long-term follow-up of the six hemispherectomized patients, especially the long-term seizure control and cognitive function after surgery, are described.

RESULTSFive patients had no seizures and one was almost seizure-free during the years after surgery. Clear P300 was obtained from every electrode in the patients. Differences of P300 between patients and normal control group had no statistical significance. And the maximum PA was at the site of electrode Pz or Cz which was consistent with that of the control group and with previous findings.

CONCLUSIONSFurther modified anatomical hemispherectomy has preferable long-term antiepileptic effects. The P300 results of the patients mean that the basic cognitive function of the patients has no difference from the control group. This reflects the plasticity of the hemisphere to some extent and increases the affirmation of the long-term curative effects of further modified anatomical hemispherectomy from both neuropsychological and neurophysiological aspects.

Adult ; Disease-Free Survival ; Event-Related Potentials, P300 ; Female ; Follow-Up Studies ; Hemispherectomy ; Humans ; Male ; Seizures ; surgery ; Treatment Outcome