To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

OBJECTIVETo investigate the expression and mutation of Mad1, Mxi1 and Rox genes in leukemia cells.

METHODSExpression and mutation of Mad1, Mxi1 and Rox genes in bone marrow mononuclear cells (BMMNC) from 26 de novo acute leukemia (AL) patients, and in peripheral blood mononuclear cells (PBMNC) from 30 healthy volunteers, as well as in 7 human leukemic cell lines were analyzed by reverse transcription-polymerase chain reaction (RT-PCR), single strand conformational polymorphism (SSCP) and DNA sequencing.

RESULTSRT-PCR showed that all the above cells expressed Mad1, Mxi1 and Rox mRNA. SSCP revealed four polymorphisms: two in Mad1, one each in Mxi1 and Rox. DNA sequencing detected nine missense mutations: two in Mad1 in AL patients, four in Mxi1 (three in AL patients and one in KG-1 cell line), and three in Rox in AL patients. The mutations of Mad1, Mxi1 and Rox mRNA were detected in 2, 3 and 3 patients, respectively.

CONCLUSIONIt is for the first time to demonstrate the mutations of Mad1, Mxi1 and Rox genes in AL patients suggesting these mutated genes involve in the pathogenesis of leukemia.

Adolescent ; Adult ; Aged ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; Female ; Humans ; Leukemia ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; metabolism ; Polymorphism, Single-Stranded Conformational ; Repressor Proteins ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the expression of heat shock protein 90 (HSP90) on the cell surface of highly invasive human prostate cancer cells PC3 and its possible molecular mechanisms of its effect on cell invasion through analyzing FAK/Src signaling pathway.

METHODSThe expression of cell surface HSP90 on PC3 cells was studied by immunofluorescence staining and surface biotinylation assay respectively. A specific HSP90 antibody was used to inhibit the cell surface HSP90. In vitro cell invasion was assessed by modified Boyden chambers. Phosphorylated FAK on tyr 397, 576, 577 and 925, and phosphorylated c-Src on tyr 416 were examined by Western blot assay. The association between FAK and c-Src was analyzed by immunoprecipitation. The effects of FAK knockdown by siRNA or Src kinases inhibitor PP2, with or without anti-HSP90 antibody, on PC3 cell invasion were also evaluated.

RESULTSA pool of HSP90 was detected on the cell surface of PC3 cells. A specific HSP90 antibody significantly retarded tumor cell invasion. Concomitant with this finding, targeting cell surface HSP90 significantly inhibited the phosphorylations of FAK and c-Src, and also the interactions between FAK and c-Src. FAK knockdown or PP2 dramatically suppressed cell invasion, however, anti-HSP90 antibody didn't further inhibit cell invasion.

CONCLUSIONSCell surface HSP90 promotes human prostate cancer cell invasion through a FAK/c-Src signaling, with may be a novel therapeutic target against metastatic tumors.

Antibodies ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Focal Adhesion Protein-Tyrosine Kinases ; genetics ; metabolism ; Gene Knockdown Techniques ; HSP90 Heat-Shock Proteins ; immunology ; metabolism ; Humans ; Male ; Neoplasm Invasiveness ; Phosphorylation ; Prostatic Neoplasms ; metabolism ; pathology ; Pyrimidines ; pharmacology ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transfection ; src-Family Kinases ; antagonists & inhibitors ; metabolism

Objective To optimize the oxygen therapy regimens for infants with pulmonary diseases during bronchoscopy.Methods A prospective randomized,controlled,and single-center clinical trial was conducted on 42 infants who underwent electronic bronchoscopy from July 2019 to July 2021.These infants were divided into a nasal cannula(NC)group and a modified T-piece resuscitator(TPR)group using a random number table.The lowest intraoperative blood oxygen saturation was recorded as the primary outcome,and intraoperative heart rate and respiratory results were recorded as the secondary outcomes.Results Compared with the NC group,the modified TPR group had a significantly higher level of minimum oxygen saturation during surgery and a significantly lower incidence rate of hypoxemia(P<0.05).In the modified TPR group,there were 6 infants with mild hypoxemia,2 with moderate hypoxemia,and 1 with severe hypoxemia,while in the NC group,there were 3 infants with mild hypoxemia,5 with moderate hypoxemia,and 9 with severe hypoxemia(P<0.05).The modified TPR group had a significantly lower incidence rate of intraoperative respiratory rhythm abnormalities than the NC group(P<0.05),but there was no significant difference in the incidence rate of arrhythmias between the two groups(P>0.05).Conclusions Modified TPR can significantly reduce the risk of hypoxemia in infants with pulmonary diseases during electronic bronchoscopy,and TPR significantly decreases the severity of hypoxemia and the incidence of respiratory rhythm abnormalities compared with traditional NC.

OBJECTIVEElevation of reactive oxygen species (ROS), especially the level of superoxide is a key event in many forms of cardiovascular diseases. To study the mechanism of tea polyphenols against cardiovascular diseases, we observed the expressions of ROS-related enzymes in endothelial cells.

METHODSTea polyphenols were co-incubated with bovine carotid artery endothelial cells (BCAECs) in vitro and intracellular NADPH oxidase subunits p22phox and p67phox, SOD-1, and catalase protein were detected using Western blot method.

RESULTSTea polyphenols of 0.4 microg/mL and 4.0 microg/mL (from either green tea or black tea) down-regulated NADPH oxidase p22phox and p67phox expressions in a dose-negative manner (P < 0.05), and up-regulated the expressions of catalase (P < 0.05).

CONCLUSIONSTea polyphenols regulate the enzymes involved in ROS production and elimination in endothelial cells, and may be beneficial to the prevention of endothelial cell dysfunction and the development of cardiovascular diseases.

Animals ; Camellia sinensis ; chemistry ; Carotid Arteries ; cytology ; Catalase ; biosynthesis ; Cattle ; Cells, Cultured ; Down-Regulation ; Endothelial Cells ; drug effects ; enzymology ; metabolism ; Flavonoids ; isolation & purification ; pharmacology ; Membrane Transport Proteins ; biosynthesis ; NADPH Dehydrogenase ; biosynthesis ; NADPH Oxidases ; Phenols ; isolation & purification ; pharmacology ; Phosphoproteins ; biosynthesis ; Polyphenols ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; biosynthesis ; Superoxide Dismutase-1 ; Up-Regulation

Chuangxinmycin (CM) from Actinoplanes tsinanensis was an antibiotic discovered by Chinese scientists about 40 years ago. It contains a new heterocyclic system of indole fused with dihydrothiopyran, whose biosynthetic mechanism remains unclear. CM is used as an oral medicine in the treatment of bacterial infections in China. The simple structure makes CM as an attractive candidate of structure modification for improvement of antibacterial activity. Recently, we analyzed the secondary metabolites of Actinoplanes tsinanensis CPCC 200056, a CM producing strain, as a natural CM analogue. We discovered the first natural CM analogue 3-demethylchuangxinmycin (DCM) as a new natural product. Compared to CM, DCM exhibited a much weaker activity in the inhibition of the bacterial strains tested. The finding provides valuable information for the structure-activity relationship in the biosynthesis of CM.
Anti-Bacterial Agents ; chemistry ; isolation & purification ; China ; Indoles ; chemistry ; isolation & purification ; Micromonosporaceae ; chemistry ; Structure-Activity Relationship

Previous studies demonstrated that interleukin-12 (IL-12) enhances the non-MHC-restricted cytotoxic activity of NK cells and facilitate specific allogeneic human cytotoxic T lymphocyte responses against fresh leukemia cells and cell lines. The Wilms' tumor gene, WT1 mRNA, has been used as a marker of minimal residual disease (MRD) for evaluating therapeutic efficacy of patients with leukemia or myelodysplastic syndrome (MDS). This study was aimed to investigate whether in vitro IL-12 can lower WT1 gene expression in peripheral blood monuclear cells (PBMNC) from patients with leukemia or MDS. PBMNC from these 30 patients and 5 healthy volunteers were cultured at 5 x 10(5) cells/ml alone with or without 100 units/ml of IL-12 for 3 days. WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR) since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia and MDS. The results demonstrated that WT1 mRNA in PBMNC of 5 healthy volunteers was less than 10(3) copies/microg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 10(4.8) to 10(4.2) copies/microg of total RNA in 6 CML patients, from 10(5.4) to 10(4.8) copies/microg in 12 MDS patients and from 10(5.0) to 10(4.2) copies/microg in 5 AML patients in CR, but not reduced in 5 of 7 AML in non-CR. It is concluded that IL-12 significantly decrease the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 may be of considerable benefit in the elimination of MRD in patients with hematological malignancies.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Interleukin-12 ; pharmacology ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; WT1 Proteins ; biosynthesis ; genetics

[Objective]To assess the effects of high flow nasal cannular insufflation(HFNCI)on preoxygenation and extension of safe apneic period during tracheal intubation.[Methods]Patients were randomly allocated into facemask(FM),facemask plus HFNCI(FM+HFNCI),HFNCI and HFNCI plus nasopharyngeal airway(HFNCI+NPA) groups. Facemask was adopted in FM and FM+HFNCI groups,while HFNCI was used in HFNCI and HFNCI+NPA groups for preoxygenation. All patients except for those in FM group received HFNCI during tracheal intubation. PaO2, SaO2,HR and MAP were recorded and analyzed.[Results]There was no significant difference in PaO2and SaO2after preoxygenation among groups(P>0.05). During apneic tracheal intubation period,PaO2decreased significantly in FM group while increased in HFNCI+NPA group. The Δ PaO2in FM group(Mean value was -5.4 kPa)was significantly bigger than those in other groups(Mean values in FM+HFNCI,HFNCI,and HFNCI+NPA groups were -0.5,-0.8 and 1.4 kPa,respectively(P < 0.001). All values at the success of tracheal intubation were much above the safe limits.[Conclusion]HFNCI provides effective preoxygenation and may extend safe apneic period in patients with patent airway.

OBJECTIVETo evaluate the aesthetic outcome after application of rectangle periosteous flap to cover corrugator.

METHODSOn the basis of regular rhytidectomy, a inter-eyebrow periosteous flap is applied to cover the fascia flap of corrugator by means of turnover. This manipulation helps to separate the skin from the muscle and prevent the re-adherence between the skin and muscle.

RESULTSThe approach was applied on 15 cases. The follow-up ranged from 6 months to 1 year. The results were satisfactory.

CONCLUSIONSThe method is effective in elimination of inter-eyebrow crease.

Adult ; Female ; Humans ; Middle Aged ; Muscle, Skeletal ; transplantation ; Periosteum ; transplantation ; Reconstructive Surgical Procedures ; methods ; Rhytidoplasty ; methods ; Skull ; Surgical Flaps ; Treatment Outcome

OBJECTIVE(99m)Tc-[bis (dimethoxypropylphosphinoethyl)-ethoxyethylamine (PNP5)]-[bis (N-ethoxyethyl)-dithiocarbamate (DBODC)] nitride ([(99m)Tc (N) (PNP5) (DBODC)](+)) is a new myocardial perfusion tracer with high heart uptake and rapid liver clearance. The objectives of this study were to compare the myocardial imaging of [(99m)Tc (N) (PNP5) (DBODC)](+) with (99m)Tc-MIBI in a canine model of acute myocardial ischemia.

METHODSThe left anterior descending artery (LAD) was occluded in 12 adult beagle dogs, adenosine was then infused intravenously at a rate of 0.14 mg.kg(-1).min(-1) for 6 min. At the end of 3 min of adenosine infusion, 185 MBq of [(99m)Tc (N) (PNP5) (DBODC)](+) or (99m)Tc-MIBI was injected intravenously. The occluder was released after 6 min adenosine infusion. Serial myocardial SPECT imaging acquisitions were obtained at 0.5, 1, 1.5 and 2 h after tracer injection, respectively. Rest myocardial SPECT imaging was acquired in the next day.

RESULTSSimilar as (99m)Tc-MIBI, [(99m)Tc (N) (PNP5) (DBODC)](+) exhibited high heart uptake, minimal lung uptake and minimal redistribution. No significant myocardial washout was observed with both tracers over a period of 2 hours. [(99m)Tc (N) (PNP5) (DBODC)](+) clearance from the liver was more rapid than that with (99m)Tc-MIBI (heart-liver radio at 60 min, 1.36 +/- 0.43 vs. 0.58 +/- 0.21, P = 0.005). [(99m)Tc (N) (PNP5) (DBODC)](+) the ability to detect myocardial ischemia was also comparable between the two tracers ([(99m)Tc (N) (PNP5) (DBODC)](+) detected 3.60 +/- 1.52 defect segments, (99m)Tc-MIBI detected 4.25 +/- 0.96 defect segments, P = 0.48). The image quality of [(99m)Tc (N) (PNP5) (DBODC)](+) was better than (99m)Tc-MIBI.

CONCLUSION[(99m)Tc (N) (PNP5) (DBODC)](+) is comparable to (99m)Tc-MIBI on detecting myocardial ischemia in this model and liver clearance is more rapid and the image quality is better than (99m)Tc-MIBI.

Animals ; Disease Models, Animal ; Dogs ; Male ; Myocardial Ischemia ; diagnostic imaging ; Organophosphorus Compounds ; Organotechnetium Compounds ; Technetium Tc 99m Sestamibi ; Tomography, Emission-Computed, Single-Photon