Objective To evaluate the effects of volume resuscitation with different fluids on expression of aquaporin 1 (AQP-1) in pulmonary capillary endothelial cells of endotoxemic rats.Methods Fifty pathogen-free male Sprague-Dawley,rats,weighing 250-300 g,aged 6-8 weeks,were randomly divided into 4 groups (n =10 each):control group (group C),lipopolysaccharide group (group LPS),NaCl group (group NS),6％ hydroxyethyl starch 130/0.4 group (group HES) and hypertonic hydroxyethyl starch 40 group (group HSH).Sepsis was induced with LPS 5 mg/kg injected via the femoral vein.At l0 min after the model was successfully established,the corresponding fluids were infused into the femoral vein at a constant speed (15 ml/kg over 6 h) via a pump.At 0,3 and 6 h after LPS administration,blood samples were collected from the femoral artery for measurement of serum tumor necrosis factor-α (TNF-α) concentrations and for blood gas analysis.PaO2 and lactate (Lac) concentrations were recorded and oxygenation index (PaO2/FiO2) was calculated.The rats were sacrificed at 6 h after LPS administration,and lungs were removed for determination of AQP-1 expression in pulmonary capillary endothelial cells and wet-to-dry lung weight ratio (W/D ratio),and for microscopic examination of the pathological changes which were scored.Results Compared with group C,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly increased,AQP-1 expression was down-regulated,and PaO2 and oxygenation index were decreased in LPS,NS,HES and HSH groups.Compared with group LPS,W/D ratio,serum TNF-a and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in NS,HES and HSH groups.Compared with group NS,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in HES and HSH groups.Compared with group HES,the blood Lac concentrations were significantly decreased,and no significant changes were found in the other parameters in group HSH.Conclusion Volume resuscitation with NaC1,hydroxyethyl starch 130/0.4 and hypertonic hydroxyethyl starch 40 can reduce the lung injury effectively in endotoxemic rats,and hypertonic hydroxyethyl starch 40 provides more significant efficacy,and up-regulated AQP-1 expression in pulmonary capillary endothelial cells is involved in the mechanism.

A young female complained repeated nasal discharge for over three months with discomfort of right cheek, and oral antibiotics had less effect. She has a history of "root canal therapy" five years before. Physical examination found purulent secretion in the right middle nasal meatus, and light tenderness in the right side of the maxillary sinus area. The CT scan of paranasal sinus shown possible fungal infection of right maxillary sinus. Finally the nasal endoscopic surgery confirmed the fungus ball of right maxillary sinus with foreign body (the root canal filling material).
Endoscopy ; Female ; Foreign Bodies ; diagnosis ; Humans ; Maxillary Sinus ; microbiology ; Maxillary Sinusitis ; microbiology ; Mycoses ; diagnosis ; Respiratory Tract Infections ; microbiology ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of midazolam on human ether-a-go-go (hERG) K+ channels exogenously expressed in human embryonic kidney cells (HEK-293) and the underlying molecular mechanisms.

METHODSWhole-cell patch clamp technique was used to record WT, Y652A and F656C hERG K+ current expressed in HEK-293 cells.

RESULTSMidazolam inhibited hERG K+ current in a concentration-dependent manner, the half-maximum block concentrations (IC50) values were (1.31 ± 0.32) µmol/L. The half-activation voltage (V1/2) were (2.32 ± 0.38) mV for the control and (-1.96 ± 0.83) mV for 1.0 µmol/L midazolam. The half-inactivation voltage (V1/2) was slightly shifted towards negative voltages from (-49.25 ± 0.69) mV in control to (-57.53 ± 0.53) mV after 1.0 µmol/L midazolam (P < 0.05). Mutations in drug-binding sites (Y652A or F656C) of the hERG channel significantly attenuated the hERG current blockade by midazolam.

CONCLUSIONMidazolam can block hERG K+ channel and cause the speed of inactivation faster. Mutations in the drug-binding sites (Y652 or F656) of the hERG channel were found to attenuate hERG current blockage by midazolam.

Dose-Response Relationship, Drug ; Ether-A-Go-Go Potassium Channels ; drug effects ; HEK293 Cells ; Humans ; Midazolam ; pharmacology ; Mutation ; Patch-Clamp Techniques ; Potassium Channel Blockers ; pharmacology

Objective To clone IRX9 gene from Dendrobium huoshanense and perform the bioinformatics and codon bias analysis. Methods The full-length cDNA sequence was cloned from the IRX9 gene based on transcriptome sequencing data of D. huoshanense generated in our previous study by using RT-PCR and further analyzed by bioinformatic methods. Results The cloned IRX9 gene was 1 533 bp, containing a 1 047 bp open reading frame (ORF) which encoded 348 amino acids. The theoretical molecular weight was 39 350 and its isoelectric point was 7.26. IRX9 protein was a hydrophilic protein and had no signal peptide, which had multiple phosphorylation sites and glycosylation sites and might be located in the plasma membrane. Phylogenetic analysis indicated that sequence of amino acids had the highest homology with D. candidum and the conserved region “DXD” of the GT-A superfamily. Codon bias analysis showed that IRX9 gene prefered to use A/T ending codon, with 27 skewed codons. Nicotiana is the most suitable host for exogenous expression of IRX9 gene. Conclusion The IRX9 gene was cloned from Dendrobium huoshanense successfully and it provided a theoretical basis for regulating the growth and development of D. huoshanense and improving the yield and quality of D. huoshanense.

Brucellacapt is a new serological test based on immunocapture-agglutination technique that has been used in many countries ,but there was no related article reported in China .The value of the Brucellacapt method in diagnosing human brucellosis was evaluated in this study .Among 120 suspected patients ,75 patients and 45 negatives were diagnosed by SAT and RBPT method combination with their clinical symptoms .Sera from all 120 people were tested by the method of Brucella-capt and iELISA ,and the results were ,consequently ,analyzed and compared .It showed that sensitivity ,specificity ,consis-tency rate ,Kappa value ,and area under ROC curve were found to be 82 .7% ,88 .9% ,85 .0% ,0 .69 ,and 0 .86 ,respectively for Brucellacapt ,whereas they were found to be 90 .7% ,64 .4% ,80 .8% ,0 .57 ,and 0 .78 for iELISA .In conclusion ,specific-ity ,consistency rate ,Kappa value ,and area under ROC curve were higher in Brucellacapt method than that in iELISA .How-ever ,the sensitivity of iELISA is higher .

OBJECTIVETo investigate effects of dahuang zhechong pill ( DHZCP) on the cell cycle and the related signal pathways in vascular smooth muscle cells (VSMCs) stimulated by platelet-derived growth factor (PDGF) with the method of serum pharmacology.

METHODSDNA synthesis in VSMCs was examined by detecting 5'-bromo-2'-deoxyuridine incorporation with the immunocytochemical method. The cycle of VSMCs was evaluated with flow cytometry. Expressions of cyclin D1, p27, protein kinase Cα (PKCα), and phosphorylated extracellular signal regulated kinase 1/2 (ERK1/2) were quantified by Western blot method.

RESULTSDHZCP containing serum significantly inhibited DNA synthesis of PDGF-stimulated VSMCs, arrested the cells in G G(1) phase, modulated the protein expressions of cyclin D D(1) and p27, and suppressed the activation of PKCα and ERK1/2.

CONCLUSIONDHZCP containing serum inhibits VSMCs proliferation via modulating the expressions of cell cycle proteins to arrest the cell in G G(1) phase, which is attributed to, at least in part, suppressing PKCα-ERK1/2 signaling in VSMCs.

Animals ; Aorta, Thoracic ; cytology ; Blood Proteins ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; DNA ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; G1 Phase ; drug effects ; physiology ; MAP Kinase Signaling System ; physiology ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; enzymology ; Platelet-Derived Growth Factor ; pharmacology ; Protein Kinase C-alpha ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To assess the effectiveness of puncturing Sifeng points (EX-UE10) and pricking Back-Shu points in treating dyspepsia due to chemotherapy for triple-negative breast cancer (TNBC). Method Sixty patients were randomized into an observation group (30 cases) and a control group (30 cases). The observation group was intervened by puncturing Sifeng points and pricking Back-Shu points, once a week. The selected Back-Shu points included bilateral Pishu (BL20), Weishu (BL21) and Geshu (BL17). The control group was treated by promoting gastrointestinal motility (itopride hydrochloride 50 mg) and supplementing digestive enzymes (compound azintamide tablets). The two groups were observed before and after treatment in terms of traditional Chinese medicine (TCM) symptom score, nutritional status score and Karnofsky Performance Score (KPS). The therapeutic efficacies were also assessed. Result The total effective rate was 93.3％ (28/30) in the observation group versus 70.0％ (21/30) in the control group, and the between-group difference was statistically significant (P<0.05). The TCM symptom score showed significant improvement in both groups after treatment (P<0.01), and the improvement in the observation group was more significant than that in the control group (P<0.01). After treatment, the score of Patient Generated-Subjective Global Assessment (PG-SGA) decreased significantly in both groups (P<0.01), while there was no significant difference in the score between the two groups (P>0.05). The KPS score increased significantly in both groups after treatment (P<0.01), and there was a significant difference between the two groups (P<0.05), indicating a more significant improvement of KPS score in the observation group. Conclusion Puncturing Sifeng plus pricking Back-Shu points is effective in treating dyspepsia due to chemotherapy for TNBC. It can improve patient's appetite and quality of life.

OBJECTIVETo investigate and compare the effects and mechanisms of three functional parts of Dahuang Zhechong Pill (DHZCP), including drugs with the function of removing blood stasis and promoting blood circulation (FP-I), drugs with the function of expelling heat and moistening dryness (FP-II), and drugs with the function of nourishing yin and replenishing blood (FP-III) of DHZCP, on platelet-derived growth factor (PDGF)-stimulated vascular smooth muscle cells (VSMCs) proliferation with the method of serum pharmacology.

METHODSVSMCs proliferation of rat was assayed by measuring the cell viability with the 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) method. DNA synthesis in VSMCs was examined by detecting 5'-bromo-2'-deoxyuridine incorporation with the immunocytochemical method. Cycle of VSMCs was evaluated with flow cytometry. Expression of cyclin D1, p27, PKCα, and phosphorylated extracellular signal regulated kinase 1/2 (ERK1/2) was quantified by the Western blotting method.

RESULTSThe FP-I and FP-III containing serum was capable of inhibiting PDGF-stimulated proliferation and DNA synthesis of VSMCs, arrested VSMCs in G phase, downregulated cyclin D1, and upregulated p27 expression (P <0.01 or P <0.05). The FP-I and FP-III containing serum also inhibited the PDGF-induced phosphorylation of tyrosine of ERK1/2 and PKCα expression (P <0.01 or P <0.05).

CONCLUSIONSFP-I and FP-III of DHZCP are able to inhibit VSMCs proliferation via interrupting PKCα-ERK1/2 signaling, modulating the expression of cell cycle proteins to result in arresting the cells in G phase. The inhibitory effect is mainly related to the function of removing blood stasis and promoting blood circulation, slightly to the function of nourishing yin and replenishing blood, but not to the function of expelling heat and moistening dryness.

Animals ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; DNA ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Activation ; drug effects ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; MAP Kinase Signaling System ; drug effects ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Platelet-Derived Growth Factor ; pharmacology ; Protein Kinase C-alpha ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum ; metabolism

Bacterial Typing Techniques ; methods ; Brucella ; classification

OBJECTIVETo study the interaction among aquaporin1 (AQP), aquaporin2 (AQP2) and antisecretory factor( AF) , and their expression in the rat inner ear for furthur understanding of Meniere' s disease.

METHODSInner ear tissue section of six healthy male Sprague-Dawley rats was performed and Envision immunochemical staining was applied to detect the expression of AF, AQP1 and AQP2 in the rat inner ear. Vestibular and cochlear tissues of twenty healthy male Sprague-Dawley rats were dissected. Coimmunoprecipitation and Western Blot were used to specifically immunoprecipitate AF protein in the vestibular and cochlear tissues with monoclonal antibodies against AQP1 and polyclonal antibodies antibodies against AQP2 to detect the above precipitate with specific antibodies against AF.

RESULTS(1) AF was widely distributed in the inner ear, such as marginal cells of stria vascularis , five classes of spiral ligament fibrocyte , Reissner's membrane, basilar membrane, ampullar crest and so on with mild or moderate staining. In addition, round membrane was moderately or markedly stained. Positive immunostaining was found in the cochlear spiral ganglion, vestibular nerve and cochlear nerve. AQP1 was distributed in the intermediate cells in stria vascularis, type III fibrocyte of spiral ligament, basilar membrane and round membrane with moderate to marked degree of immunostaining intensity. AQP2 was mainly localized to the type II, IV, and V fibrocyte of spiral ligament, with moderate to marked degree of immunostaining intensity, round membrane was weakly stained. (2) No band was observed in the control and a single immunoreactive band of 60 000 was observed, which was equal to the molecular mass of AF.

CONCLUSIONS(1) AF, AQP1 and AQP2 have its individual specific localization in the rat inner ear, which was close to the parts of endolymph, so regulating water of the endolymph may be possible. (2) The range of localization of AF overlapped the distribution of AQP1 and AQP2. The results showed the existence of AF protein in the immunoprecipitate using co-immunoprecipitation combined with Western Blot. It suggested that the interaction between AQP1, AQP2 and AF might be possible.

Animals ; Aquaporin 1 ; metabolism ; Aquaporin 2 ; metabolism ; Cochlea ; metabolism ; Ear, Inner ; metabolism ; Male ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley