Objective To investigate the genotypes of aminoglycoside-modifying enzymes (AMEs) produced by Pseudomonas aeruginosa isolated from Huaibei Miner's General Hospital.Methods Minimum inhibitory concentration of 36 strains of P. aeruginosa to 3 aminoglycoside antibiotics was determined. AME genes were amplified by polymerase chain reaction (PCR). Results The P. aeruginosa isolates were resistant to multiple antibiotics. Most (62.2% to 81.1%) were resistant to aminoglycosides. And 27 strain (75.0%) carried one or more types of AME genes, including ant(3″)-Ⅰ (63.9%), aac(6')-Ⅱ (58.3%), aac(6')-Ⅰ (50.0%), aac(3)-Ⅱ (38.9%) and ant(2″)-Ⅰ (36.1%). The aac(3)-Ⅰ, aac(3)-Ⅲ, aac(3)-Ⅳ, aph(3')-Ⅳ genes were not identified.Conclusions The study indicates that the P. aeruginosa isolates in Huaibei are multi-resistant to aminoglycoside antibiotics. The prevalence of AMEs-positive strains is high.

Objective To explore a safe and efficient method for endotracheal intubation in mice.Methods 60 BALB/c mice were random divided into 2 groups, direct intubation under direct vision group, ( n = 30) and direct intubation under light by transillumination group ( n = 30).After successful tracheal intubation at the first judgment, mouse received a tracheal instillation of 3 mg/kg lipopolysaccharide.Bronchoalveolar lavage (BAL) was then performed three times with 0.8 ml sterile saline.Exudate cells were centrifuged and stained with Wright's fluid.Whether the intubation indeed succeeded was judged by the amount of neutrophil infiltrated into the lungs.The mortality after intubation, the time consumed for each successful tracheal intubation and the times of attempting for intubation in two groups were assessed to approve the efficiency of two techniques.Results Eight vs.two mice died within 24 hours in two groups respectively.The time consumed for each successful tracheal intubation and success rate for the first time for two groups were (92.6 ±23.4)s vs (64.0±20.1)s and 53.3% vs 86.7% respectively, which the consumed time was significantly different, while the final success rate almost the same for the two groups.Conclusion Both direct intubation under direct vision and intubation under light by transillumination can successfully intubate the tube into the trachea.But direct intubation under direct vision by transillumination is more efficient and safe in endotracheal intubation in mice.

Objective To clarify whether oxymatrine ( OM) could suppress the activation of pancreatic stellate cells ( PSC) and explore the potential molecular mechanism .Methods The proliferation of PSC line LTC 14 being activated by TGF-β1 with OM treatment at different concentrations (OM group) was measured. SOD level was determined by ELISA and p 38-MAPK mRNA was determined by real-time PCR.Results The proliferation of PSC in the control group , 0.1, 0.5, 1, 2, 5 g/L OM group was (1.51 ±0.08), (1.50 ± 0.07), (1.15 ±0.04), (1.15 ±0.04), (1.08 ±0.06), and (1.08 ±0.10), respectively.The level of the control group was lower than the groups where the concentration of OM reached or exceeded 0.5mg/ml ( all P=0.000).SOD level of LTC 14 cells in the control group, TGF-β1 group, 0.5 and 1 g/L OM group was (0.087 ±0.005), (0.073 ± 0.004), (0.085 ± 0.010), and (0.086 ± 0.007), respectively. No statistically significant difference existed among the groups (P=0.095).The p38-MAPK mRNA expression of PSC in the control group, TGF-β1 group, 0.5, and 1 g/L OM group was (1.000 ±0.000), (1.979 ± 0.505), (0.606 ±0.111), and (0.303 ±0.159), respectively.The p38-MAPK mRNA level of TGF-β1 group was higher than that of the control group (P=0.002), and that of 0.5 mg/ml OM group and 1 mg/ml OM group was lower that of TGF-β1 group ( P=0.000 ) , while no statistical difference was found between 0.5 mg/ml OM group and 1 mg/ml OM group.Conclusions OM could suppress the activation of PSC in vitro and the suppression of p38-MAPK mRNA expression may be involved .

Objective To investigate the chemical constituents of red algae Hypnea charoides. MethodsThe constituents were isolated by column chromatography and their structures were elucidated by chemical properties and spectroscopic analyses. Results Three compounds were isolated and their structures were identified as 3, 6-dimethyl-8-(4-methyleneheptan-3-yloxy) octane-1-amine (Ⅰ), palmitate-K (Ⅱ), and (2R, 1′S, 2′S, 3′R)-N-(1′-hydroxymethyl-2′, 3′-dihydroxy-heptadecenyl)-2-hydroxy-12, 13-methylene-tetracosanamide (Ⅲ). Conclusion Compounds Ⅰ and Ⅲ are new compounds named as charoidesine and charoidesamide.

Objective To find new compound from gorgonian.Methods Three ceramides have been isolated from the South China Sea gorgonian Echinogorgia sp.by silica gel column chromatography.Results Their structures were established as(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl)]-hexadecanamide(1),(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl)]-heptadecanamide(2),and(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl]-octadecanamide(3) by spectroscopic methods and chemical conversion.Conclusion It is the first time to report the three chemical compounds from coral Echinogorgia sp.and compound 2 is a new compound.

OBJECTIVE To investigate the genes associated with the drug-resistance to ?-lactam antibiotics in Pseudomonas aeruginosa(PAE) isolated from clinical patients in Huaibei,Anhui Province.METHODS Antibiotic sensitivity test was performed by VITEK-AMS.The three-dimensional method was taken to differentiate various beta-lactamases.The ?-lactamases and the outer membrane protein D2(oprD2) genes were detected by polymerase chain reaction(PCR) in 36 PAE strains.RESULTS In 36 PAE strains,the positive rates of genes of TEM,DHA,CTX-Ge and CARB were 77.8%,69.2%,27.8% and 25.0%,respectively,that of the OXA-1,OXA-10,VIM-2 and IMP-1 were 16.7%,16.7%,13.9% and 2.8%,respectively.The rate of lacking oprD2 gene was 58.3%,and none possessed VEB,SHV,GES,PER,GIM and SPM-2 genes.CONCLUSIONS P.aeruginosa carries various beta-lactamase genes in clinical patients of Huaibei,and the deletion ratio of oprD2 gene is higher.

OBJECTIVETo explore the effect of microneedle combined with Lauromacrogol on skin capillary network.

METHODS24 male Leghone (1.5-2.0 kg in weight) were randomly divided into three groups as group A (microneedle combined with Lauromacrogol), B (microneedle combined with physiological saline) , and C(control). The cockscombs were treated. The specimens were taken on the 7th, 14th, 21th , and 28th day postoperatively. HE staining, immunohistochemical staining and special staining were performed for study of the number of capillary and collagen I/III , as well as elastic fibers.

RESULTSThe color of cockscombs in group A became lightening after treatment. The number of capillary decreased as showing by HE staining. The collagen I and III in group B was significantly different from that in group A and C (P < 0.05). Special staining showed proliferation of elastic fibers in group B.

CONCLUSIONSIt indicates that microneedle combined with Lauromacrogol could effectively reduce the capillary in cockscomb without any tissue fibrosis. Microneedle can stimulate the proliferation of elastic fiber, so as to improve the skin ageing process.

Animals ; Capillaries ; anatomy & histology ; Chickens ; Comb and Wattles ; blood supply ; drug effects ; Male ; Needles ; Polyethylene Glycols ; pharmacology ; Punctures ; instrumentation ; methods ; Random Allocation ; Skin Aging

Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.

Objective To explore the influence of LPS treatment on related molecules in Smads and ERK1/2 signal pathway in pancreatic stellate cell line LTC-14.Methods LTC-14 cells were cultured in vitro, and were treated with LPS at different dose in different time points.Protein expressions of related molecules in Smads pathway and ERK1/2 pathway and α-SMA in LTC-14 Cells were examined by Western blot.Results On Treated LTC-14 cells by 0, 1, 5, 10, 20 and 50 mg/L LPS,protein expressions of Smad3 were 0.15±0.02, 0.37±0.02, 0.44±0.01, 0.46±0.02, 0.372±0.01 and 0.24±0.03;expressions of Smad7 were 0.79±0.05, 0.84±0.02, 0.55±0.03, 0.45±0.03, 0.34±0.02 and 0.92±0.07;p-ERK1/2 levels were 0.48±0.05, 0.74±0.03, 0.72±0.04, 0.89±0.02, 0.81±0.02 and 0.72±0.03;p-cPLA2 levels were 0.15±0.03, 0.30±0.01, 0.31±0.01, 0.30±0.02, 0.28±0.03 and 0.32±0.02;α-SMA levels were 0.56±0.06, 0.62±0.06, 0.54±0.04, 1.03±0.11, 1.39±0.08 and 1.28±0.10.The changes of protein expressions before and after LPS treatment were obvious (all P<0.01).The protein expressions of ERK1/2 were 0.56±0.03, 0.57±0.02, 0.53±0.02, 0.58±0.02, 0.59±0.05 and 0.55±0.04, which did not change obviously along with increased LPS dosages.LTC-14 cells treated with 10 mg/L LPS for 0, 1, 3, 6 and 9 h,the expressions of Smad3 were 0.69±0.05, 0.68±0.07, 1.02±0.14, 1.82±0.0 and 2.04±0.11,those of Smad7 were 2.77±0.10, 1.37±0.08, 1.45±0.14, 0.78±0.09 and 0.63±0.06,those of p-ERK1/2 were 0.16±0.03, 0.32±0.05, 0.79±0.03, 1.50±0.07 and 1.77±0.04,those of p-cPLA2 were 0.15±0.04, 0.32±0.06, 0.63±0.04, 0.95±0.04 and 1.49±0.10,those of α-SMA were 0.84±0.03, 1.26±0.21, 1.81±0.19, 4.28±0.26 and 4.37±0.15, all of which changed obviously as the treatment time increased (P<0.05 or 0.01).The expressions of ERK1/2 were 0.75±0.03, 0.72±0.02, 0.80±0.04, 0.74±0.03 and 0.85±0.09, which did not change obviously as the treatment time increased.Conclusions LPS could upregulate the expression of α-SMA in a time-and dose-dependent way, and activate intracellular Smads and ERK1/2 inflammatory pathways, which may be the potential molecular mechanism of the development of chronic pancreatitis.

Objective To measure the mRNA expression of interleukin receptors (IL-2R?IL-4R and IL-10R) in peripheral blood mononuclear cells (PBMCs) from patients with chronic idiopathic urticaria (CIU). Methods Thirty CIU patients and 30 controls were enrolled in this study. PBMCs were separated from the peripheral blood specimens. Reverse transcription-polymerase chain reaction (RT-PCR) technique was applied to semi-quantitatively analyze the mRNA expression. Results The expression level of IL-10R mRNA was significantly increased in patients with CIU than that in the healthy controls, while that of IL-2R and IL-4R mRNA in PBMCs showed no significant difference. Conclusion Our results suggest that IL-10R might be involved in the pathogenesis of CIU.