Objective To investigate the influence of abaI expression on acinetobacter baumannii biofilm formation.Methods Acinetobacter baumannii strain S isolated from bums patients was collected for the study,while the standard strain ATCC19606 was served as the control.At 6,24 and 48 hours,the gene expressions of abaI,pgaA,pgaB and pgaC were detected by real-time fluorescent quantitative PT-PCR,secretion of N-acyl-homoserine lactones (AHLs) by biological sensor and biofilm formation by MTT method.Results (1) Gene expressions of abaI,pgaA,pgaB and pgaC at 6 hours were 8.63 ±5.93,1.98 ± 1.93,1.01 ± 1.32 and 2.67 ± 3.46 respectively,which showed a quick increase at 24 hours (22.81 ± 17.60,5.13 ± 4.32,5.66 ± 3.97,11.97 ± 7.75 respectively),followed by a rapid decline in 48 hours (3.43 ± 0.88,1.30 ± 0.24,3.01 ± 3.00,3.02 ± 3.29 respectively).Gene expressions of pgaB and pgaC at 6 hours and that of pgaA and pgaC at 48 hours revealed statistically significant differences from those at 24 hours (P ＜ 0.05).(2) AHLs showed a level of 18.49 ± 11.03 at 6 hours,reached a peak of 52.23 ± 15.95 at 24 hours,then descended to 5.53 ± 0.94 at 48 hours.AHLs level at 24 hours showed statistically significant difference from that at 6 hours and 48 hours (P ＜ 0.05).(3)Biofilm formation at 24 hours and 48 hours was 2.83 ±0.44 and 2.71 ±0.15 respectively,far higher than that at 6 hours (0.49 ± 0.11,P ＜ 0.05).(4) In the correlation analysis among AHLs,biofilm formation and gene abaI,pgaA,pgaB and pgaC expressions,significant positive correlation was found between abaI and pgaA and between AHLs and pgaC expression (P ＜ 0.05).Conclusion Acinetobacter baumannii may regulate gene expressions of pgaA and pgaC responsible for biofilm formation to adjust to the external environment by means of changing abaI gene expression and AHLs secretion.

Animals ; Central Nervous System Diseases ; drug therapy ; Drug Design ; Drugs, Chinese Herbal ; Humans ; Inflammation ; drug therapy

OBJECTIVETo investigate the clinical effect and safety of chiropractic in treating cervicogenic sudden hearing loss.

METHODSFrom January 2011 to October 2013, 90 patients with cervicogenic sudden hearing loss were randomly divided into treatment group and control group according to the random number table produced by SPSS 19.0 software. In the treatment group, there were 17 males and 28 females, aged from 31 to 62 years old with an average of (47.57±9.43) years; course of disease was from 1 to 3 days with an average of (1.43±0.68) days; pure-tone audiometry score was from 46.5 to 77.8 dB with the mean of (61.20±9.83) dB; Northwick Park Neck Pain Questionnaire (NPQ) score was from 17 to 31 scores with an average of (23.46±7.18) scores. In the control group, there were 15 males and 30 females, aged from 28 to 64 years old with an average of (45.77±6.99) years; course of disease was from 1 to 3 days with an average of (1.50±0.73) days; pure-tone audiometry score was from 48.1 to 75.0 dB with the mean of (63.91±8.05) dB; Northwick Park Neck Pain Questionnaire (NPQ) score was from 20 to 29 scores with an average of (25.61±10.43) scores. The patients of control group were treated with dexamethasone intravenous drip of 10 mg, 3 days later, decreased to 5 mg, 3 days again. And with the methycobal intravenous drip of 500 μg, treatment continued for 10 days. The patients of treatment group were treated with chiropractic additionally except for the therapeutic methods of control group. Chiropractic included local muscle loosening, attacking point, bilateral pulling atlanto-axial joint, and continuous treatment for 10 days. The pure-tone audiometry score and NPQ score were compared between two groups after treatment.

RESULTSAfter the treatment, pure-tone audiometry score and NPQ score in treatment group improved to (40.23± 8.14) dB and (12.70±8.29) scores respectively, which were obviously better than that of control group's (37.70±10.61) dB and (21.24±11.13) scores (P<0.05).

CONCLUSIONCompared with routine method for cervicogenic sudden hearing loss, additional chiropractic can improve hearing and relieve neck pain effectively.

Cervical Vertebrae ; Female ; Hearing Loss, Sudden ; therapy ; Humans ; Male ; Manipulation, Chiropractic ; methods ; Medicine, Chinese Traditional ; Middle Aged

OBJECTIVETo explore the analgesic effect of Zhixin Formula (ZF) and its effects on spinal glial fibrillary acidic protein (glial fibillary acidic protein, GFAP, a marker of astrocyte) , CD11b (a maker of microglia), Toll like receptors (TLR2 and TLR4) and nuclear factor κB (NF-κB) in bone cancer pain model rats.

METHODSTotally 20 male SD rats were randomly divided into the blank control group and the bone cancer pain group, 10 in each group. The bone cancer pain model was induced by injecting ascites tumor fluid containing 3 x 10(3) Walker256 cell line from the left tibia. Ethological tests, X-ray test, and HE staining were performed to confirm a successful modeling. After model was successfully established, 70 male SD rats were randomly divided into seven groups, 10 in each group: the blank control group, the bone cancer pain group (as the model group), the Western medicine (WM) group (Tramadol Hydrochloride), the high dose ZF group, the middle dose ZF group, the low dose ZF group, and the Chinese medicine (CM) group (Wulin Zhitong Capsule). Fourteen days after modeling, rats in the high, middle, and low dose ZF groups were administrated by gastrogavage with 9, 4.5, and 2.25 g/kg ZF water condensed preparation respectively, once a day for seven consecutive days. On day 21 MS typical protein expressions including GFAP, CD11b, TLR (2,4) and NF-κB from cornu dorsal medullae spindis L4-L5 were detected by immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.

RESULTSCompare with the blank control group, increased weight in the model group was slow and showed a decreasing trend (P < 0.01), spontaneous ambulatory pain score (SAPS) obviously increased (P < 0.01), paw withdrawal threshold (PWT) obviously decreased in the model group (P < 0.05, P < 0.01). Results of lateral tibial X-ray and HE staining showed obvious changes and damage occurred in bone structures of rats in the model group. Immunohistochemistry showed that GFAP expression significantly increased in the model group (P < 0.05). Protein levels of NF-κB also significantly increased in the model group (P < 0.05). Compared with the model group, CD11b expressions obviously decreased in the middle and high dose ZF groups (P < 0.01). Meanwhile, protein expressions of TLR2 and TLR4, as well as NF-κB also obviously decreased (P < 0.05).

CONCLUSIONZF had analgesic effect, which might be probably related to inhibiting proliferation and activation of gliocytes, as well as activation of TLRs and NF-κB.

Animals ; Astrocytes ; Bone Neoplasms ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Glial Fibrillary Acidic Protein ; Male ; Microglia ; NF-kappa B ; metabolism ; Osteosarcoma ; Pain ; Rats ; Rats, Sprague-Dawley

Background Bulbar conjunctiva tissue appears to be thinning,elasticity declined,tension reduced and fascia atrophied in conjunctivochalasis.Histopathological examination of conjunctivochalasis shows decrease of elastic fibers and melt of collagen fibers.But there are fewer studies on the ultrastructure of conjunctiva of conjunctivochalasis up to now.Objective This study was to investigate the ultrastructure change of conjunctiva tissue in conjunctivochalasis.Methods Five loose conjunctiva samples of conjunctivochalasis and 5 normal conjunctival tissue samples were collected and ultra-microstructure changes of these samples were observed under the transmission electron microscope.Results The number of fibroblasts in conjunctivochalasis lamina was progressively decreased.The shape of fibroblasts was long and fusiform.Somatic synapse was slim.Nucleus-cytoplasm ratio was increased.Disorder,scattered and broken of the collagen fibril were seen,and some areas were dissolved or lacunae.Normal conjunctival fibroblasts were oval or polygonal,with wieners and long somatic synapse,and intercellular matrix was full of collagen fibril and dense arranged fibers.Fibroblasts in fascia of eonjunctivochalasis were linear,and collagen fibril was seriously defected.Fascia fibroblasts of normal bulbar conjunctiva were spindleshaped and bigger than conjunctivochalasis fibroblasts.There were full of collagen fibrils in intercellular matrix.Conclusions The collagen fibril is decreased and fibroblast cells are degenerated in lamina and fascia of conjunctivochalasis.

Objective To do screening acute lymphoblastic leukemia patients scFv antibody single chain variable region to cre-ate conditions for the expression and obtain further specificity of antibody fragments.Methods In this study,patients with newly diagnosed acute lymphoblastic leukemia serum as coating antigen using phage display technology,screening phage an-tibody specificity from the semi-synthetic human phage antibody libraries,the first to target the immune antigen-coated tab-let,phage library was added,so that with the target antigen-specific binding phage antibody was immobilized on plates immu-nization,could not be specifically bound phages were rinsed.The eluted specific binding phage,E.coli infection.Could get the specific antibody gene containing phagemid.Results After three “adsorption-elution-amplification”screening process,got stronger leukemia patient antigen-specific phage antibody variable region fragment and identification.Conclusion Got better strain affinity antibody fragments,to create the conditions for the next fragment expression,identification and clinical appli-cation.

Objective: To explore the peri-operative treatment and 3 years post-operative outcome in patients with rheumatic mitral valve stenosis and small left ventricle. Methods: A total of 152 patients with rheumatic mitral valve stenosis and severe pulmonary hypertension were studied. According to left ventricular end diastolic index (LVEDI) value, the patients were divided into 2 groups: Small left ventricle (Small) group, the patients with LVEDI value≤60 mm, n=67 and Non-small left ventricle (Non-small) group, the patients with LVEDI value>60 mm,n=85. Peri-operative condition and follow-up parameters were compared between 2 groups. Results: Compared with Non-small group, Small group had increased pre-operative CVP, SPAP and decreased LVEDV, LVEDVI and RVEF at admission, all P<0.05. All patients had liver damage at admission, the indexes (except alkaline phosphatase) were higher in Small group than Non-small group, and the function was recovered prior operation by proper treatment. The immediate post-operative SPAP in Small group and Non-small group were (67.3.1±23.4) mmHg and (61.3±26.9) mmHg, P>0.05. There were 52 severe pulmonary hypertension patients received iloprost inhalation, their SPAP was decreased than non-inhalation patients at incision closing; SPAP was lower in Small group inhalation patients than Non-small group inhalation patients, all P<0.05. Post-operative LVEDVI was lower and SPAP was higher in Small group than Non-small group. At 3 years post-operation, LVEDVI, RVEF increased, NYHA classification decreased and 6MWT prolonged in both groups; while LVEDVI was lower and 6MWT was shorter in Small group than Non-small group, P<0.05. The median survival time at (3.7±1.6) years post-operation was shorter in Small group than Non-small group,P<0.05, while the mortality was similar between 2 groups (2.6% vs 2.0%),P>0.05. Conclusion: The patients with mitral valve stenosis and small left ventricle had the worse pre-operative condition and higher risk in surgery; peri-operative treatment could improve their cardiac and liver function, but they still had the shorter post-operative median survival time.

Objective To study the pharmacokinetics profiles of ibupro-fen and pseudo ephedrine with chlorpheniramine suspension after single and multiple dosing in healthy volunteers.Methods Three single and one multiple oral doses of ibuprofen and pseudo ephedrine with chlorphe-niramine suspension were given to healthy volunteers respectively.Ibu-profen concentrations in plasma were determined by HPLC-UV method.Pseudo ephedrine and chlorpheniramine concentrations in plasma were determined by HPLC-MS-MS method.The pharmacokinetic parameters were obtained with statistical analysis by DAS Ver 2.0.Results The main pharmacokinetic parameters of 3 single doses(ibuprofen:200,400,600 mg pseudo ephedrine:30,60,90 mg;chlorpheniramine:2,4,6 mg)and multiple dose(ibuprofe,pseudo ephedrine and chlorphe-niramine 200,30,2 mg,respectively)shown that the concentration-time curves of ibuprofen,pseudo ephedrine and chlorpheniramine were described by one-compartment open model and physiological disposi-tions were assumed by linear kinetics characteristics.Conclusion In multiple dosing study,physiological dispositions of ibuprofen,pseudo e-phedrine and chlorpheniramine all existed no induction and inhibition of drug enzyme phenomenon.

Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.