Based on single factor tests,the optimum vacuum belt drying conditions of Qibai Pingfei granule were obtained through Box-Benhnken central combination design and RSM. In this study, drying time, drying temperature and extract density were chosen as independent variables, while transferring rate ginsenoside Rg₁, Re, Rb₁and astragaloside IV were taken as dependent variables. The optimum parameters are as follows: drying time of 112 min, drying temperature of 87 °C and extract density of 1.30 g · mL⁻¹. At the optimum condition, transferring rate ginsenoside Rg₁+ Re, Rb₁and astragaloside IV were 88.01%, 87.31%, 84. 34%. Above all, the optimum processing parameters of vacuum belt drying of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Temperature ; Vacuum

To study the anti-proliferation effect of lambda-carrageenan oligosaccharides (lambda-CO) on human umbilical vein endothelial cells (HUVECs) and expression of apoptotic relevant genes, the influence of lambda-CO on HUVECs proliferation was measured by MTT assay; apoptotic rate, cell cycle distribution and the level of active caspase-3 of HUVECs were analyzed using flow cytometry; the mRNA level of apoptosis related genes was determined by RT-PCR. At a high concentration of 1 mg x mL(-1), lambda-CO significantly inhibited the endothelial cell proliferation. Annexin-V FITC/PI double stain assay showed that when treated with 0, 0.8, 1 mg x mL(-1) of lambda-CO for 24 h, cell apoptotic rates were (1.67 +/- 1.6)%, (11.48 +/- 2.4)% and (13.81 +/- 2.2)%, respectively, when treated for 48 h, cell apoptotic rates were (2.02 +/- 2.3)%, (13.84 +/- 1.9)% and (38.72 +/- 2.5)%, respectively, cell cycle assay showed the decrease of cells in G0/G1 phase, and increase in S phase. Furthermore, we observed the level of active caspase-3 increased in a dose-dependent manner at 24 th and 48 th. RT-PCR results indicated that mRNA of TNFalpha, p53, caspase-8 and caspase-3 in cells increased after treated with lambda-CO. lambda-CO induce apoptosis of HUVECs in a dose-dependent way and arrests cells at S phase, which mainly due to the up-regulation of apoptotic genes such as TNFalpha, p53, caspase-8, caspase-3 and increase the level of active caspase-3.
Angiogenesis Inhibitors ; pharmacology ; Apoptosis ; drug effects ; Carrageenan ; pharmacology ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; biosynthesis ; genetics ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Humans ; Oligosaccharides ; pharmacology ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Umbilical Veins ; cytology

2-(4-Methylthiazol-5-yl) ethyl nitrate hydrochloride (W1302) is a nitro containing derivative of clomethiazole, which is a novel neuroprotective agent with both carbon monoxide (NO) donor and weak γ-aminobutyric acid type A (GABA_A) receptor allosteric regulatory excitatory effect. The current study used a rat model of transient middle cerebral arteryocclusion (tMCAO) brain injury to evaluate the therapeutic effect of W1302 on ischemic stroke and explore its potential mechanisms of action. This experiment has been reviewed and approved by the Laboratory Animal Management and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences (ethical review forms No. 620, 632, 5013). The results showed that gavage administration of 1, 3, and 10 mg·kg^-1 of W1302 can significantly reduce the volume of cerebral infarction in rats with ischemia for 2 h and reperfusion for 24 h, and the therapeutic effect was better than that of 200 mg·kg^-1 of DL-3n-butylphthalide. W1302 significantly increased NO levels in blood and brain tissue. It increased cerebral blood flow and brain adenosine triphosphate (ATP) content after reperfusion, as well as, inhibited the expressions of inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue. The time window of W1302 was between 120-180 min after ischemia. These research results demonstrated that W1302 could increase NO release, dilate blood vessels, and increase cerebral blood flow, improve energy supply to brain tissue and increase ATP levels, and inhibit neuro-inflammation, which playing the protective roles in tMCAO stroke model rats. This provides theoretical support for the clinical application of W1302 in the treatment of ischemic stroke.

BACKGROUND: Three-dimensional finite element analysis has been used by many scholars from department of orthopedics, but the results of postoperative evaluation of hip preserving treatment for osteonecrosis of femoral head are different. OBJECTIVE: To study the biomechanical changes of the femoral head and the biomechanical changes of the proximal femur after greater trochanter bone flap for the treatment of femoral head necrosis using three-dimensional finite element method, and to verify the mechanical safety and effectiveness. METHODS: One case of unilateral femoral head necrosis in ARCOIII stage undergoing parallel vascularized greater trochanter bone flap transplantation was selected. Computed Tomography data of proximal femur were collected before and 6 months after the operation, and preserved in DICOM format. With the aid of computer technology, professional medical modeling software, MIMICS and HYPERMESH, were used to establish the three-dimensional geometric models of the proximal femur. These models were divided into normal group, necrosis group and repair group. Finite element analysis software ANSYS was utilized to simulate human body standing and movement in different situations. The model was divided by free mesh, and given material parameters to establish normal proximal femur, femoral head necrosis and bone defect. Greater trochanter bone flap was applied in repairing three-dimensional finite element model of bone defect. Loads were loaded on different finite element models. The maximum displacement of the femoral head and the stress distribution in the proximal femur of the three groups were observed under different loading models. RESULTS AND CONCLUSION: (1) Under the same load, the maximum displacement of the three sets of models was 0.61 mm in the normal group, 0.66 mm in the necrosis group, and 0.61 mm in the repair group, respectively. Maximum Von Mises stress was greater in necrosis model than in the normal molding. The maximum Von Mises stress gradually decreased in the repair model, and was close to normal value. (2) Three groups of models showed stress concentration above the rotor in femoral neck region. The maximum stress in the trochanteric position was higher in necrosis models than in normal models. The maximum stress in this region gradually increased after repair, but was still lower than the failure stress of bone. (3) The results confirm that the maximum stress and the maximum displacement are closer to the normal value after greater trochanter bone flap for treatment of osteonecrosis of the femoral head. The greater trochanter is safe and reliable for repairing bone defect of femoral head.

OBJECTIVETo explore the clinical value of ultrasonic surface localization in internal jugular vein catheterization.

METHODSTotally 150 patients with American Society of Anesthesiologists physical status I -III who were planning to receive elective surgeries were randomized into anatomical landmark group, ultrasonic surface positioning group, and ultrasound-guided group using computed random table, with 50 cases in each group. The right internal jugular vein catheterization was performed after tracheal intubation. In the anatomic landmark group, patients were punctured using surface marks through central approach. In ultrasonic surface positioning group and ultrasound-guided group, patients were punctured with ultrasonic localization and guidance through central approach. The relationship between internal jugular vein and carotid artery, the position of the needle into the vein, the success rate of puncture, the change times of puncture point, and the complications were recorded.

RESULTSUltrasound scan revealed that the relationship between the right internal jugular vein and the right common carotid artery could be divided into three types: parallel (12.7%), partial overlapping (69.3%), and complete overlapping (18.0%). The average "safety distance" of jugular vein puncture was (1.15 +/- 0.47) cm. The success rate of the first puncture attempt in ultrasonic surface positioning group and ultrasound-guided group were 78.0% and 82.0%, respectively, which was significantly higher than that in anatomic landmark group (22.0%) (P < 0.05), whereas the complication incidence in anatomic landmark group (12.0%) were significantly higher than those in ultrasonic surface positioning group (0) and ultrasound-guided group (0) (P < 0.05).

CONCLUSIONSUltrasonic surface positioning applied during internal jugular vein catheterization is helpful to reveal the inner diameters as well as the origin and course of arteries and veins in the puncture and identify the abnormalities as early as possible. As a simple support technique for internal jugular vein puncture, it is suitable for clinical application.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Catheterization, Central Venous ; methods ; Female ; Humans ; Jugular Veins ; diagnostic imaging ; Male ; Middle Aged ; Ultrasonography ; Young Adult

OBJECTIVETo investigate the presence of β-amyloid peptide (Aβ) deposition in the cerebellum and the expression of related miRNAs in the cerebellum of a mouse model of Alzheimer disease.

METHODSTwelve 12-month-old APPswe/PSδE9 double transgenic mice and 12 wild-type C57 mice were sacrificed and the brain tissues were taken for examination. The right hemisphere was stained with Congo red to observe the deposition of amyloid substances, and from the left hemisphere, the hippocampus and the cerebellum were dissected for detecting the expression of miRNA-135a-5p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p using real-time PCR.

RESULTSCongo red staining revealed the presence of Aβ deposition in both the hippocampus and the cerebellum of the transgenic mice but not in the control mice. Real-time PCR showed a significantly lower expression of the 4 miRNAs in the hippocampus in the transgenic mice than in the control mice (P<0.05). The expression of miRNA-135a-5p, miRNA-298-5p, and miR-669f-3p in the cerebellum was significantly lower in the transgenic mice than in the control mice (P<0.05). The expression of miRNA-298-5p and miR-669f-3p in the hippocampus was significantly lower than that in the cerebellum of the transgenic mice (P<0.05).

CONCLUSIONβ deposition also occurs in the cerebellum of APPswe/PSδE9 double transgenic mice, and its formation might be related to the down-regulation of miRNA-135a-5p, miRNA-298-5p, and miR-669f-3p.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Cerebellum ; metabolism ; Disease Models, Animal ; Hippocampus ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; MicroRNAs ; metabolism

OBJECTIVETo observe the changes of miRNA expression profiles in APPswe/PSδE9 transgenic mice and explore the possible roles of miRNA in the pathogenesis of Alzheimer's disease.

METHODSUsing miRNA chip technique, we examined the miRNA expression in the brain tissue of 6-month-old APPswe/PSδE9 transgenic mice, with age-matched wild-type mice as the control group.

RESULTSTwelve miRNAs showed differential expressions by more than two folds in APPswe/PSδE9 transgenic mice, namely miRNA-135a, miRNA-135a-2*, miRNA-298, miRNA-466b-3p, miR-669-3p, miR-142-5p, miR-144, miR-466f-3p, miR-466g, miR-200a, miR-200b and miR-96. Five miRNAs were significantly down-regulated in the transgenic mice, including miRNA-135a, miRNA-135a-2*, miRNA-298, miRNA-466b-3p, and miR-669-3p.

CONCLUSIONThe 5 down- regulated miRNA may play important roles in the pathogenesis of AD in APPswe/PSδE9 transgenic mice.

Alzheimer Disease ; genetics ; metabolism ; Animals ; Disease Models, Animal ; Gene Expression Profiling ; Mice ; Mice, Transgenic ; MicroRNAs ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; Transcriptome

OBJECTIVETo detect the expression of miRNA-135a-5p, miRNA-135a-2-3p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p in the brain tissue of the APPswe/PS δE9 double transgenic mouse model of Alzheimer's disease using real-time PCR.

METHODSSix-month-old APPswe/PS δE9 double transgenic mice and wild-type C57 mice of the same species were examined for the expressions of miRNA-135a-5p, miRNA-135a-2-3p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p in the brain tissue using real-time PCR.

RESULTSThe relative expression levels of the 5 miRNAs in the transgenic versus the wild-type mice were 0.73∓0.27 vs 1.08∓0.58, 2.47∓6.15 vs 1.65∓0.67, 0.72∓0.14 vs 1.31∓0.73, 0.57∓0.34 vs 1.06∓0.35, and 0.63∓0.26 vs 1.02∓0.18, respectively, showing significance differences in the expressions of miRNA-135a-5p, miRNA-298-5p, miRNA-466b-3p, and miR-669f-3p between the two groups (P<0.05).

CONCLUSIONSmiRNA-135a-5p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p are expressed differentially in APPswe/PS δE9 double transgenic mice, suggesting their important roles in the pathogenesis of Alzheimer disease.

Alzheimer Disease ; genetics ; metabolism ; Animals ; Brain ; metabolism ; Disease Models, Animal ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; MicroRNAs ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; methods

BACKGROUNDAcinetobacter baumannii is one of the main gram-negative bacilli in clinical practice. Nosocomial infections caused by multi-drug resistance Acinetobacter baumannii is very difficult to treat. This study was designed to investigate the antimicrobial resistance characteristics and four resistant gene expressions of aminoglycoside-modifying enzymes including N-acetyltransferases and O-phosphotransferases in Acinetobacter baumannii.

METHODSBacterial identification and antimicrobial susceptibility test were performed by Phoenix system in 247 strains of Acinetobacter baumannii. Minimal inhibitory concentrations (MICs) of seven aminoglycosides including gentamicin, amikacin, kanamycin, tobramycin, netilmicin, neomycin and streptomycin in 15 strains of multi-drug resistant Acinetobacter baumannii were detected by agar dilution. Four aminoglycoside-modifying enzyme genes were amplified by polymerase chain reaction (PCR) and verified by DNA sequencer.

RESULTSThe resistance rates of 247 strains of Acinetobacter baumannii against cefotaxime, levofloxacin, piperacillin, aztreonam, tetracycline, ciprofloxacin and chloramphenicol were more than 50%. Imipenem and meropenem showed high antibacterial activities with resistance rates of 3.2% and 4.1%. MIC50 and MIC90 of gentamicin, amikacin, streptomycin and kanamycin in 15 strains of multi-drug resistant Acinetobacter baumanii were all more than 1024 mg/L, and the resistance rates were 100%, 100%, 100% and 93.3%, respectively. But their resistance rates to tobramycin, netilmicin and neomycin were 86.7%, 93.3% and 46.7%, respectively. Three modifying enzyme genes, including aacC1, aacC2 and aacA4 genes, were found in 15 strains, but aphA6 had not been detected. Their positive rates were 93.3%, 20.0% and 20.0%, respectively. These three genes existed simultaneously in No.19 strain. Nucleotide sequences of aacC1, aacC2 and aacA4 genes shared 100%, 97.9% and 99.7% identities with GenBank genes (AY307113, S68058 and AY307114).

CONCLUSIONMulti-drug resistant Acinetobacter baumannii strains are rapidly spreading in our hospital, and their resistance to aminoglycosides may be associated with aminoglycoside-modifying enzyme gene expressions.

Acinetobacter baumannii ; drug effects ; enzymology ; genetics ; Aminoglycosides ; metabolism ; pharmacology ; Base Sequence ; Drug Resistance, Multiple, Bacterial ; Gene Expression Regulation, Bacterial ; Genotype ; Microbial Sensitivity Tests ; Molecular Sequence Data

OBJECTIVETo investigate the relationship between the drug resistance of Pseudomonas aeruginosa (PA) isolated from burn patients wounds and its mobile genetic elements, including plasmid, transposon, and integron.

METHODSThirty-two strains of PA were isolated from wounds exudate of hospitalized burn patients in Ningbo No. 2 Hospital. PA drug sensitivity was determined using GNS-448 drug sensitivity card and K-B tests. The genetic markers of plasmid, transposon and integron including traA, traF, tnpA, tnpU, merA, int I 1 were amplified by PCR and verified by gene sequencing.

RESULTSDrug resistant rate of 32 PA strains to gentamicin, amikacin, cefoperazone/sulbactam, ciprofloxacin was 43.7%, 32.0%, 46.8%, 49.9%, respectively. PA drug resistant rates to piperacillin, cefotaxime, ceftazidime, cefepime, aztreonam, piperacillin/tazobactam, levofloxacin, imipenem and meropenem were all above 56.0%. Seventeen out of 32 PA strains were found to carry transposon and (or) integron genetic markers. One strain was positive for both tnpA and merA, 8 strains were positive for both merA and int I 1, 1 strain was only positive for tnpA, 2 strains were only positive for merA, and 5 strains were positive for int I 1 only.

CONCLUSIONSPA isolated from burn wounds of hospitalized patients in Ningbo No. 2 Hospital is seriously drug resistant, which may relate with its high positive rate of mobile genetic elements of transposon and (or) integron.

Anti-Bacterial Agents ; pharmacology ; Burns ; microbiology ; DNA Transposable Elements ; Drug Resistance, Multiple, Bacterial ; genetics ; Humans ; Integrons ; Microbial Sensitivity Tests ; Plasmids ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification