OBJECTIVETo investigate the effect of hypoxia on the human pulmonary artery smooth muscle cells two pore domain potassium channels TASK-1 and the regulation of non-receptor tyrosine kinase c-Src in this process.

METHODSThe cultured human pulmonary artery smooth muscle cells (hPASMCs) were divided into: normal group, hypoxia 30 minute group, hypoxia 6 hours group and hypoxia 48 hour group, and hypoxia 48 hour + PP2 group, hypoxia 48 hour + PP3 group, hypoxia 48 hour + bpV group. Flow cytometry was used to analyze the cell cycle, RT-PCR and Western blot technique were carried out to detect the expression changes of TASK-1 mRNA and protein in different groups.

RESULTS(1) Cell Cycle Show: Compared with normal control group, with prolonged hypoxia, the percentages of hPASMCs in S phases of cell cycle were increased. While compared with hypoxia 48 hour group, the percentages of hypoxia 48 hour + PP2 group hPASMCs in S phases of cell cycle were decreased. The expression of TASK-1 mRNA on hPASMCs in acute hypoxia 6 hour group was increased, while the expression of TASK-1 protein on hPASMCs in the acute and chronic hypoxia group was decreased, and the expression of TASK-1 mRNA on hPASMCs in the chronic hypoxia group was decreased; After pre-incubation of a potent and selective inhibitor of the Src family of protein tyrosine kinases PP2, the expression of TASK-1 mRNA and protein in hypoxia 48 hour group was increased, however after pre-incubation of the inhibitor of the Src family of protein tyrosine phosphatase bpV, the expression of TASK-1 protein in hypoxia 48 hour group was decreased.

CONCLUSIONHypoxia promotes human pulmonary artery smooth muscle cell proliferation, and non-receptor tyrosine kinase c-Src may participate in the expression of two pore domain potassium channels TASK-1 regulated by hypoxia. Therefore, we hypothesized that TASK-1 channels and c-Src participatein the acute and chronic hypoxic human pulmonary vasoconstriction.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology ; Nerve Tissue Proteins ; metabolism ; Potassium Channels, Tandem Pore Domain ; metabolism ; Pulmonary Artery ; cytology ; RNA, Messenger ; Vasoconstriction ; src-Family Kinases ; metabolism

Objective To examine the antimicrobial susceptibility and prevalence of blaKPC gene in carbapenem-resistant Klebsiella pneumoniae (CRKP) strains isolated in Huashan Hospital,Fudan University.Methods The CRKP strains isolated in Huashan Hospital from January to December of 2014 were included in this study.The MICs of antibiotics were determined using CLSI broth dilution method.The blaKPC gene was amplified by polymerase chain reaction (PCR).Results A total of 205 CRKP strains were isolated,mainly from respiratory tract (76.1％,156/205) and urine specimens (18.5％,38/205).Antimicrobial susceptibility test indicated that CRKP isolates had higher resistance rates (85％-100％) to the antimicrobial agents except colistin (1.5％),tigecycline (0.5％),trimethoprim-sulfamethoxazole (51.0％) and amikacin (74.9％).Most (87.8％,180/205) of the CRKP strains were positive for blaKPC gene.Conclusions CRKP are mostly isolated from patients with lower respiratory tract infection and/or urinary tract infection in Huashan Hospital.The strains were highly resistant to the antibacterial agents tested except colistin and tigecycline.Production of KPC-type carbapenemase is the common mechanism of carbapenem resistance in these K.pneumoniae isolates.

In this study, hydrophilic matrix sustained release tablets of total lactones from Andrographis paniculata were prepared and the in vitro release behavior were also evaluated. The optimal prescription was achieved by studying the main factor of the type and amount of hydroxypropyl methylcellulose (HPMC) using single factor test and evaluating through cumulative release of three lactones. No burst drug release from the obtained matrix tablets was observed. Drug release sustained to 14 h. The release mechanism of three lactones from A. paniculata was accessed by zero-order, first-order, Higuchi and Peppas equation. The release behavior of total lactones from A. paniculata was better agreed with Higuchi model and the drug release from the tablets was controlled by degradation of the matrix. The preparation of hydrophilic matrix sustained release tablets of total lactones from A. paniculata with good performance of drug release was simple.
Andrographis ; chemistry ; Delayed-Action Preparations ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydrophobic and Hydrophilic Interactions ; Kinetics ; Lactones ; chemistry ; Tablets ; chemistry

OBJECTIVES: Chronic rhinosinusitis (CRS) is common disease in otorhinolaryngology and will lead to lower airway abnormality. However, the only lung function in CRS patients and associated factors have not been much studied. METHODS: One hundred patients with CRS with nasal polyps (CRSwNP group), 40 patients with CRS without nasal polyps (CRSsNP group), and 100 patients without CRS were enrolled. The difference in lung function was compared. Meanwhile, CRSwNP and CRSsNP group were required to undergo a bronchial provocation or dilation test. Additionally, subjective and objective outcomes were measured by the visual analogue scale (VAS), 20-item Sino-Nasal Outcome Test (SNOT-20), Lund-Mackay score, Lund-Kennedy endoscopic score. The correlation and regression methods were used to analyze the relationship between their lung function and the above parameters. RESULTS: The forced expiratory volume in 1 second (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75) of CRSwNP group were significantly lower than other groups (P<0.05). On peak expiratory flow, there was no difference between three groups. In CRSwNP group, FEV1 was negatively correlated with peripheral blood eosinophil count (PBEC) and duration of disease (r=–0.348, P=0.013 and r=–0.344, P=0.014, respectively), FEF25-75 negatively with VAS, SNOT-20 (r=–0.490, P=0.028 and r=–0.478, P=0.033, respectively) in CRSsNP group. The incidence of positive bronchial provocation and dilation test was lower in CRSwNP group (10% and 0%, respectively), with both 0% in CRSsNP group. The multiple linear regression analysis indicated that change ratio of FEV1 before and after bronchial provocation or dilation test were correlated with PBEC in CRSwNP group (β=0.403, P=0.006). CONCLUSION: CRS leading to impaired maximum ventilation and small airway is associated with the existence of nasal polyp. Lung function impairments can be reflected by PBEC, duration, VAS, and SNOT-20. In CRSwNP patients, PBEC is independent predictor of FEV₁ change ratio.
Bronchial Hyperreactivity ; Bronchial Provocation Tests ; Eosinophils ; Forced Expiratory Volume ; Humans ; Incidence ; Linear Models ; Lung* ; Nasal Polyps ; Otolaryngology ; Ventilation ; Vital Capacity

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

OBJECTIVETo plot a hour-specific transcutaneous bilirubin (TCB) nomogram for healthy neonates, and to evaluate its value for prediction of the risk of neonatal hyperbilirubinemia.

METHODSA total of 5,250 healthy full-term or near-term neonates (gestational age≥35 weeks, birth weight≥2 000 g) were enrolled as subjects. Their TCB values were continuously recorded for 168 hours after birth. The TCB values in the high-risk zones of three time periods, 24-48, 49-72, and 73-96 hours after birth, were used as predictors. The hour-specific TCB nomogram combined with the receiver operating characteristic (ROC) curve was used to evaluate the predictive value of hour-specific TCB nomogram for hyperbilirubinemia.

RESULTSAccording to the hour-specific TCB nomogram, the TCB value dramatically increased during 16-72 hours after birth, and the increase slowed down gradually during 72-144 hours. Finally, the curve reached a plateau after 144 hours. Particularly, the P95 of TCB had been stabilized at 96 hours. The P40, P75, and P95 peak values of TCB were 173, 217, and 248 µmol/L, respectively. For the prediction of hyperbilirubinemia, the areas under the ROC curve of TCB at 24-48, 49-72, and 73-96 hours after birth were 0.77, 0.85, and 0.87, respectively. The high-risk zones at 24-48, 49-72, and 73-96 hours after birth predicted the incidence rates of neonatal hyperbilirubinemia as 35.03%, 43.35%, and 79.95%, respectively, with positive likelihood ratios of 3.35, 4.75, and 22.70, respectively.

CONCLUSIONSThe hour-specific TCB nomogram and the division of TCB risk zones can give a satisfactory prediction of the incidence of neonatal hyperbilirubinemia. The neonate with a bilirubin level in the high-risk zone within 73-96 hours after birth is likely to have hyperbilirubinemia after 73-96 hours.

Bilirubin ; analysis ; Female ; Humans ; Hyperbilirubinemia, Neonatal ; diagnosis ; Infant, Newborn ; Male ; Neonatal Screening ; methods ; Nomograms ; ROC Curve

In order to establish HPLC fingerprint of Liuwei Dihuang soft capsule, and to provide certain reference for an quality control of it, the HPLC method was performed on an Agilent C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-0.02% trifluoroacetic acid as mobile phase, gradient elution volume flow of 1.0 mL x min(-1), column temperature was 30 degrees C, detection wavelength: 0-60 min, 238 nm, 60-70 min, 210 nm. The software for chromatographic fingerprint was applied to analysis different batches of Liuwei Dihuang soft capsule samples. Sixteen mutual peaks were selected as the fingerprint peaks in 12 samples with loganin as the reference peak, and all of the detected peaks were separated effectively. Cluster analysis (HCA) and similarity analysis (SA) were done based on data of 12 samples clustering analysis of 12 batches of samples were divided into 2 categories. Including 7 for the first class, the rest was second, similarities calculated by SA were all above 0.92, indicating a good similarity between the reference and twelve batches of samples, also, the analysis results of HCA and SA basically the same. This method is simple with good precision, repeatability and stability, and provides the basis for Liuwei Dihuang soft capsule quality control.
Capsules ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Quality Control

OBJECTIVETo evaluate the role of water channel AQP4 in NMDA-induced brain injury in mice.

METHODSIn AQP4 gene knockout (AQP4(-/-)) mice, brain injury was induced by microinjection of NMDA into the cortex. The injured area was determined by toluidine blue staining, degenerated neurons were detected by Fluro-Jade B staining, and increased blood-brain barrier (BBB) permeability was evaluated by IgG immunostaining.

RESULTCompared with wild-type mice, AQP4(-/-) mice exhibited increased cortical lesion area, aggravated neuron degeneration, and increased BBB disruption after NMDA microinjection.

CONCLUSIONAQP4 may play a protective role in NMDA-induced brain injury in mice.

Animals ; Aquaporin 4 ; genetics ; physiology ; Blood-Brain Barrier ; pathology ; Brain ; drug effects ; pathology ; Mice ; Mice, Knockout ; N-Methylaspartate ; toxicity

Background:The role of local treatment in oligometastatic prostate cancer (PCa) is gaining interest with the oligometastases hypothesis proposed and the improvement of various surgical methods and techniques.This study aimed to compare the short-term therapeutic outcomes of robotic-assisted laparoscopic radical prostatectomy (RALP) for oligometastatic prostate cancer (OPC) vs.localized PCa using propensity score matching.Methods:Totally 508 consecutive patients underwent RALP as a first-line treatment.The patients were divided into two groups according to oligometastatic state:the OPC group (n =41) or the localized PCa group (n =467).Oligometastatic disease was defined as the presence of two or fewer suspicious lesions.The association between the oligometastatic state and therapeutic outcomes of RALP was evaluated,including biochemical recurrence (BCR) and overall survival (OS).A Cox proportional hazards model was used to assess the possible risk factors for BCR.Results:Totally 41 pairs of patients were matched.The median operative time,the median blood loss,the overall positive surgical margin rate,the median post-operative hospital stays,and the post-operative urinary continence recovery rate between the two groups showed no statistical significance.The 4-year BCR survival rates of the OPC group and localized PCa group were 56.7％ and 60.8％,respectively,without a significant difference (P=0.804).The 5-year OS rates were 96.3％ and 100％,respectively (P =0.326).Additionally,the results of Cox regression showed that oligometastatic state was not an independent risk factor for BCR (P =0.682).Conclusions:Our findings supported the safety and effectiveness of RALP in OPC.Additionally,oligometastatic state and sites did not have an adverse effect on BCR independently.

OBJECTIVETo observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.

METHODSFour interference sites targeting at rat ICOS gene were designed and four pairs of oligonucleotide fragments were cloned into the pSilencer 4.1-CMV neo plasmid vectors then transfected into rat lymphocytes with cationic liposome. The expression of mRNA and protein of ICOS was detected by RT-PCR and flow cytometry. The alteration of lymphocyte proliferation ability was evaluated by mix lymphocyte reaction, and the secretion levels of IFN-gamma and IL-4 were measured by ELISA procedure.

RESULTSAfter transfection, the expression of mRNA and protein of ICOS in test groups were lower than that in control groups (P < 0.05). The ability of T-lymphocytes in proliferation was poor and the levels of IFN-gamma and IL-4 were reduced with ICOS gene shut down.

CONCLUSIONSRNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.

Animals ; Antigens, Differentiation, T-Lymphocyte ; genetics ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Genetic Vectors ; Inducible T-Cell Co-Stimulator Protein ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lymphocyte Activation ; Male ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; immunology ; metabolism ; Transfection