Objective:To study the feasibility of using sevoflurane induction in infants' removing stitches after cleft lip surgery. Methods:60 infants after cleft lip surgery were randomly divided into three groups:group K (ketamine group,n=20),group S (sevoflurane group,n=20) and group SN (sevoflurane and nitrous oxide group,n=20). Group K were given intramuscular ketamine 5 mg/kg,midazolam 0.05 mg/kg and atropine 0.01 mg/kg. Group S were induced with inhalation of 8% sevoflurane under 6 L/min oxygen. Group SN were induced with inhalation of 8% sevoflurane under 4 L/min nitrous oxide and 2 L/min oxygen. After induction,anesthesia was continued with inhalation of sevoflurane under 3 L/min oxygen for 2 min before starting removing stitches. HR and SpO2 were monitored regularly during operation. The induction time,recovery time,occurrence of head moving,complication such as respiratory depression and increased secretion were recorded. Results:Induction and recover time in group S and group SN were similar,but faster than that of group K. Head moving in group S and group SN were less than that in group K. There happened glossoptosis and increased secretion in all the three groups,but no differences were found significantly. Conclusion:Inhaled induction of sevoflurane has more rapid induction and recover compared with intramuscular ketamine,and can be used safely in infants' removing stitches after cleft lip surgery. Additional inhalation of nitrous oxide can not shorten infants' induction and recovery time than sevoflurane inhalation alone.

BACKGROUND: It is reported that the hip-conserving effect of osteonecrosis of femoral head is closely related to the retention of lateral column. The classification of China-Japan Friendship Hospital is based on the three-column structure, and the prediction accuracy of femoral head collapse is high. OBJECTIVE: To establish a three-dimensional finite element model for China-Japan Friendship Hospital classification of femoral head necrosis, and to analyze the mechanical changes of fibula implantation in each classification by finite element method, and to explore the significance of lateral column retention in hip preservation, so as to provide a basis for precise prediction of collapse of the classification. METHODS: Three groups of 11 kinds of three-dimensional finite element models of normal femoral head, China-Japan Friendship Hospital type femoral head necrosis (type M, type C, type L1, type L2, type L3) and fibula implantation were established. The finite element analysis was carried out by ANSYS software. The maximum stress, maximum displacement and load transfer mode of proximal femur were observed in each group. RESULTS AND CONCLUSION: (1) In the necrosis group, the strain was the largest, and the displacement was different due to the different types of necrosis. The displacement changes were as follows: Type M < type C < type L1 = type L2 < type L3. The displacement recovery of fibula implantation group was lower than that of the normal group, and the displacement recovery was different due to the different necrosis types. The displacement changes were as follows: Type M < type C < type L1 < type L2 < type L3. The reduction range of the displacement of the repaired necrotic femoral head gradually decreased from the lateral column to the medial column, which was lower than the maximum displacement of the normal femoral head. (2) The peak value of the stress nephogram of the loading area of the femoral head after necrosis was higher than that of the normal group. The peak value of necrotic type M was nearly normal. The peak value of necrotic type C was 74.5% higher than that of the normal group, and the peak value of necrotic type L was more than 100% higher than that of the normal group. The peak value of necrotic type M after operation was not only 14.2% lower than that before operation, but also was lower than that of the normal group. The peak value of necrotic type C after operation was 5.3% lower than that before operation, but higher than that of normal group. The peak value of necrosis type L after operation was lower than that before operation, but significantly higher than the normal level. (3) The load transfer in the normal femoral head was continuous. The conduction path was from the lateral column of the femoral head to the femoral moment. In the necrosis group, the internal load transfer of types M and C femoral head was continuous, and the conduction of type M was basically consistent with normal. The stress of type C conduction to femoral moment was reduced. The load transfer of L1, L2 and L3 type femoral head was interrupted. The stress changed in cliff type, and was unable to transmit to femoral moment, resulting in stress concentration in load area of the femoral head. A certain effective load transfer mode was reconstructed in the femoral head of the fibula implantation group, and the stress concentration at the femoral moment occurred in all types of conduction. Part of the load was transferred to the femoral moment through fibula, and the normal load transfer mode was partially restored. (4) China-Japan Friendship Hospital type fibula placement can prevent the collapse of the femoral head to a certain extent. The location and size of the necrosis area are very important. The closer the necrosis is to the lateral column, the easier it is to collapse and the more difficult it is to repair. The retention of the lateral column is an important factor for accurate prediction of the collapse of the femoral head.

Objective To investigate the risk factors for postoperative delirium in patients after vascular free flap reconstruction performed under general anesthesia.Methods Two hundred and sixteen ASA Ⅰ-Ⅲ patients aged 18-80 yr undergoing vascular free flap reconstruction surgery were enrolled in this study.Patient characteristics before and during operation were recorded.The patients were followed up for 5 days after operation.Their level of consciousness,severity of pain and sleep quality were evaluated daily.The patients were divided into 2 groups according to the occurrence of delirium during the 5 days after operation:delirium group and non-delirium group.The method of CAM-ICU was reed in the diagnosis of postoperative delirium.Multivariate logistic regression was used to analyze the risk factors for postoperative delirium.Results logistic regression analysis showed that old age,history of alcohol abuse and sleep diacrder after operation were risk factors for delirium developed after free flap surgery.Conclusion Old age,history of alcohol abuse and sleep disorder after operation were the risk factors for postoperative delirium in patients after vascular free flap reconstruction performed under general anesthesia.

AIM:To investigate the effects of DL-3-n-butylphthalidle (NBP) on angiogenesis of human um-bilical vein endothelial cells ( HUVECs) and the role of vascular endothelial growth factor ( VEGF)/VEGF receptor 2 (VEGFR2)-Notch1/Delta-like ligand 4 (Dll4) signaling pathway in this process. METHODS:The serum-free medium and anoxic tank were used to simulate the conditions of hypoxia and ischemia ( H/I). HUVECs were divided into control group, H/I group, H/I+NBPhigh group and H/I+NBPlow group. The HUVECs in control group were conventionally cul-tured, and those in H/I group were cultured under H/I intervention. The HUVECs in H/I+NBPhigh group were treated with NBP at 20 μmol/L under H/I intervention. The HUVECs in H/I+NBPlow group were treated with NBP at 5 μmol/L under H/I intervention. The cell viability of each group was measured by CCK-8 assay. The migration ability of the HUVECs in each group was detected by cell scratch test. The vessel formation ability of the HUVECs was examined by in vitro angiogenesis assay. The expression of VEGFR2, Notch1 and Dll4 at mRNA and protein levels was determined by qPCR and Western blot, and the expression of VEGF was determined by qPCR and ELISA. RESULTS:NBP increased the viability of HUVECs, and promoted the migration ability and the formation of blood vessels in vitro under H/I interven-tion. These effects of NBP at high dose were more significant than those at low dose. NBP increased the expression of VEGF, VEGFR2, Notch1 and Dll4 at mRNA and protein levels (P<0.05). CONCLUSION:NBP promotes HUVECs to form blood vessels under H/I intervention. The mechanism may be related to the activation of VEGF/VEGFR2-Notch1/Dll4 signaling pathway.

BACKGROUNDMycoplasma pneumoniae (M. pneumoniae) is one of the common pathogens causing atypical pneumonia. In recent years, resistance to macrolides has become more common, especially in China. Previous studies have confirmed that the mutation at position 2063 in domain V of the 23S rRNA is the most prevalent, followed by the mutation at position 2064. Reported molecular detection methods for the identification of these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction (PCR) with high-resolution melt analysis, and nested PCR-linked with capillary electrophoresis, etc. The most commonly used method for monitoring resistance-conferring mutations in M. pneumoniae is direct DNA sequencing of PCR or nested PCR products. However, these methods are time-consuming, labor-intensive or need expensive equipments. Therefore the development of rapid and sensitive methods is very important for monitoring the resistance globally.

METHODSIn this study, we reported a fast and cost-effective method for detecting 2063 and/or 2064 macrolide resistant mutations from specimens using a modified allele-specific PCR analysis, and all results were compared with the sequencing data. We also analyzed the clinical courses of these samples to confirm the modified allele-specific PCR results.

RESULTSAmong 97 M. pneumoniae specimens, 88 were found to possess mutations by this method, and all modified allele-specific PCR analysis results were consistent with the sequencing data. The data of the clinical courses of these 97 cases showed that they suffered from severe pneumonia. Erythromycin showed better efficacy on cases from which no macrolide resistance mutation was found on their specimens. However, in some cases from which mutations were detected, erythromycin monotherapy had poor efficacy, and on these patients severe symptoms improved only when azithromycin was added to the treatment.

CONCLUSIONSThe drug-resistant M. pneumoniae is very common in Beijing, China. Our modified allele-specific PCR analysis can identify erythromycin resistant mutations more rapidly from specimens than any other method currently available. Erythromycin is still effective for treating patients infected with the mutation negative M. pneumoniae, but this treatment fails to work on mutant organisms. This method can facilitate clinicians in selecting appropriate therapy within short timescales.

Alleles ; Anti-Bacterial Agents ; pharmacology ; China ; Drug Resistance, Bacterial ; genetics ; Erythromycin ; pharmacology ; Mycoplasma pneumoniae ; drug effects ; genetics ; Polymerase Chain Reaction ; methods

An expression plasmid carrying anthrax protective antigen (PA) gene was constructed, which has an OmpA signal sequence attached to the 5' end of PA gene. The plasmid was transformed into E. coli and induced to express recombinant PA (rPA) . The recombinant protein, about 10% of the total bacterial protein in volume, was secreted to the periplasmic space of the cell. After a purification procedure including ion-exchange, hydrophobic interaction chromatography, and gel filtration, about 15 mg of 95 % pure rPA was obtained from 1-liter culture. The bioactivity of rPA was proved by in vitro cytotoxicity assay. The polyclonal antiserum from rabbits immunized with rPA could inhibit the action of anthrax lethal toxin in vitro, which suggests that antibodies against rPA can provide high passive protection against anthrax. The results reported here may be helpful to develop a safe and efficacious recombinant PA vaccine against anthrax.
Amino Acid Sequence ; Animals ; Anthrax Vaccines ; immunology ; Antigens, Bacterial ; chemistry ; genetics ; immunology ; toxicity ; Bacterial Toxins ; chemistry ; genetics ; immunology ; toxicity ; Base Sequence ; Mice ; Molecular Sequence Data ; Plasmids ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVEUsing doubly labeled water method to validate the colmputer science application's activity monitor (CSA) in assessing physical activity of free-living adults in Beijing, in order to develop equations to predict total daily energy expenditure (TEE) and activity related energy expenditure (AEE) from activity counts (AC) and anthropometric variables.

METHODSA total of 72 healthy adults (33 males and 39 females, mean age 43.6 +/- 4.0 yr) were monitored for 7 consecutive days by CSA. TEE was simultaneously measured using doubly labeled water method. Average AC (counts/min(-1)) was compared with TEE, AEE and physical activity level (PAL).

RESULTSPhysical activity determined by AC was significantly related to data on energy expenditures: TEE (r = 0.31, P < 0.01), AEE (r = 0.30, P < 0.05), and PAL (r = 0.26, P < 0.05). Multiple stepwise regression analysis showed that TEE was significantly influenced by gender, fat-free mass (FFM) or BMI and AC (R(2) = 0.52 - 0.70) while AEE was significantly influenced by gender, FFM and AC (R(2) = 0.25 - 0.32).

CONCLUSIONAC from CSA activity monitor seemed a useful measure in studying the total amount of physical activity in free-living adults while AC significantly contributed to the explained variation in TEE and AEE.

Activities of Daily Living ; Adult ; Anthropometry ; Body Weight ; Calorimetry, Indirect ; Energy Metabolism ; physiology ; Female ; Humans ; Male ; Monitoring, Physiologic ; instrumentation ; Motor Activity ; physiology ; Physical Fitness ; physiology

OBJECTIVETo investigate the expression profiles of myocardin gene during the differentiation of bone marrow-derived mesenchymal stem cell to smooth muscle cells in the conditional medium combined with a high concentration of fetal bovine serum (FBS).

METHODSMarrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myocardin and several smooth muscle cells marker genes were determined by immunofluorescence, RT-PCR and Western blot before and 3, 7, 10, 14 d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

RESULTSNaive bone marrow-derived mesenchymal stem cells displayed multiple morphological forms including fusiform, polygon, oval, and micro-spherical, as compared to the single macro-spindle form after the induction. Typical appearance of peak valley was displayed on the 21st day after induction. At the same time, the expression of smooth muscle marker genes was reinforced along with an up-regulation of myocardin expression. Immunofluorescence study showed that the cells expressing myocardin and smooth muscle marker genes such as alpha-SMA and SM-MHC increased. Fluorescence domain of myocardin translocated from cytoplasm to nucleus and the amounts of double positive cells for myocardin with alpha-SMA or SM-MHC also increased. RT-PCR confirmed that the mRNA expression of myocardin increased gradually and remained stabilized after achieving its peak on the 7th day after induction. The expression of smooth muscle marker genes, alpha-SMA and SM22alpha, remained stable on the 10th day of induction. It was also confirmed by Western blot that the protein expression of both myocardin and alpha-SMA were markedly increased during the induction. Finally, transmission electron microscopy revealed the presence of myofilament on the 21st day after induction.

CONCLUSIONSBone marrow-derived mesenchymal stem cells can be effectively induced into smooth muscle-like cells by conditioned medium combined with 20% FBS. Myocardin plays an important role in the differentiation process of bone marrow-derived mesenchymal stem cells to the peripheral smooth muscle cells.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Cattle ; Cell Differentiation ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; physiology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; physiology ; Up-Regulation

OBJECTIVETo compare the knee osteoarthritis (OA) models in rabbits by different concentrations of papain and provide data for exploring pathogenesis and treatments of this disease.

METHODSSixty New Zealand white rabbits were randomly divided into four groups of 15 each and given injections into the right knee on days 1, 3 and 5 including intra-articular injections of 2%, 5% or 10% (w/v) papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg (experimental groups). The 0.9% NaCl (w/v) with a dose of 0.1 ml/kg were injected intra-articularly into the right knees of rabbits in the control group. The rabbits were sacrificed at 2, 4, 6 weeks respectively after the initiation of papain injection and these OA models were evaluated through recording the width of knee joint, performing the morphological observation and histological evaluation of articular cartilage and synovium.

RESULTSThe degenerative changes were demonstrated in knee joints of rabbit in all experimental groups, such as thinner articular cartilage, fibrillation and destroyed cartilage matrix, and inflammation, proliferation, and degeneration of the synovial tissue. All these changes were much worse with increased concentration and prolonged observation time.

CONCLUSIONDifferent severity of OA are established through intra-articular injections of 2%, 5% or 10% papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg. These models are of the characters of short period and a good reproducibility.

Animals ; Disease Models, Animal ; Male ; Osteoarthritis, Knee ; chemically induced ; pathology ; Papain ; toxicity ; Rabbits

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism