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Objective:To establish a rapid method to detect the minimum inhibitory concentration (MIC) of imipenem in Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) based on ompK36 gene′s GD mutation. Methods:This was a methodological evaluation study. A total of 258 isolates of Klebsiella pneumoniae were collected from Lishui Municipal Central Hospital from March 2011 to December 2019. Porin gene ompK36 and carbapenemase genes blaKPC, blaNDM, blaIMP and blaOXA-48 were amplified by PCR and confirmed by sequencing. The MIC was detected and confirmed by microbroth dilution susceptibility test, and the corresponding patterns of genotype and MIC were constructed. Based on the patterns, a method for rapid detection of imipenem MIC by real-time fluorescence PCR (RT-PCR) was designed and established. The 159 isolates of non-repetitive Klebsiella pneumoniae collected by Lishui Disease Prevention and Control Center (CDC) from 2017 to 2019 were used for further verification. The sensitivity and specificity were calculated by fourfold table. Kappa test was used to compare the consistency between RT-PCR and microbroth dilution susceptibility test. Results:Among 258 isolates, 109 isolates did not carry carbapenemase gene, 65 isolates carried ompK36 gene GD mutation, 127 isolates carried blaKPC, 15 isolates carried blaNDM, 9 isolates carried blaIMP, and blaOXA-48 was not detected. With mircobroth dilution susceptibility test as the standard, there were 3 corresponding patterns between the drug resistance gene and the imipenem MIC of Kp: when all the 4 carbapenemase genes were negative, MIC≤1 mg/L, the sensitivity was 100% (107/107) and the specificity was 98.4% (125/127); when blaKPC was positive and ompK36 gene GD mutation was negative, 4 mg/L≤MIC≤16 mg/L, the sensitivity was 88.2% (60/68) and the specificity was 98.8% (164/166); when blaKPC and ompK36 gene GD mutation were both positive, MIC≥32 mg/L, the sensitivity was 96.6% (57/59) and the specificity was 96.6% (169/175). RT-PCR detected blaKPC, blaNDM, blaIMP, blaOXA-48 genes accurately.The RT-PCR results of ompK36 gene GD mutation in the KPC-producing isolates were 100% consistent with the sequencing results. In the 159 isolates from Lishui CDC, the sensitivity and specificity of imipenem MIC detected by RT-PCR were higher than 95% in all 3 patterns with mircobroth dilution susceptibility test as the standard, and Kappa value was 0.971. Conclusion:The RT-PCR based on ompK36 gene GD mutation was helpful to quickly determine the MIC range of imipenem in KPC-Kp.
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Objective:To evaluate the best parameter of predicting the operation time and clearance of flexible ureteroscopic lithotripsy through comparing correlations between three stone burden parameters (diameter, area, volume) and the operation time or clearance retrospectively.Methods:Clinical data and CT images of 70 patients who performed flexible ureteroscopic lithotripsy because of single kidney stone in our center from January 2018 to December 2019 were retrospectively reviewed. There were 46 males and 24 females; their age was (47±12) years old. Stones were located on the left side in 28 cases and right side in 42 cases; 32 cases in the renal pelvis , 29 cases in the lower calyx, 6 cases in the middle calyx and 3 cases in the upper calyx. The free software ITK-SNAP 3.6.0 to segment kidney stones in 3D models with the CT image was used. The stone volume was calculated automatically after the segment. The largest section of the stone on the CT coronal plane was selected to measure the maximum length (D) and width (d) of the stone, the maximum diameter of the stone was D, and the stone area was calculated using the formula 0.25πDd. The patients were divided by the operation clearance into total clearance group and partial clearance group. The correlations between three stone burden parameters (volume, diameter, area) and operation time or clearance of the flexible ureteroscopic lithotripsy were compared. Simple linear regression model was also applied to compare three measurement methods. Then other factors which may affect the operation time was evaluated with the stepwise linear regression model, such as stone component and location.Results:The median operation time was 63(50, 84)min. Of 70 cases, 47 cases were in the stone-free group, with stone volume 633(248, 1 087)mm 3, maximum diameter 15(10, 19)mm, and area 82(49, 186)mm 2. 23 cases were in the non stone-free group, with volume 696(408, 1 418)mm 3, maximum diameter 15(12, 20)mm, area 105(73, 201)mm 2. There was no difference between the two groups in volume, maximum diameter and area of stones (all P>0.05). The stone-free rate of the diameter >2 cm group was 55% (6/11), ≤2 cm group was 70% (41/59). There was no significant difference between the two groups. Correlation between stone volume and operation time is the best. The correlation coefficient of stone volume is 0.58, of stone diameter is 0.33, of stone area is 0.34. Coefficients of determination of the stone volume linear regression is the best, too. R square of stone volume is 0.36, of stone diameter is 0.17, of stone area is 0.22. Forward stepwise regression model shows stone volume is the most important parameter which correlate with operation time. None of stone volume, diameter or area has significant correlation with the clearance of stone. Conclusion:Stone volume is the best predictive parameter of the stone burden because it has the best correlation with the operation time of the flexible ureteroscopic lithotripsy of the single kidney stone.
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Objective@#To analyze the molecular epidemiological characteristics of Escherichia coli producing NDM-5 carbapenemase in the neonatal department of our hospital. @*Methods@#Three carbapenem-resistant Escherichia coli strains(E1, E2 and E3) isolated from neonatal ward of our hospital from August to September of 2017 were collected. Vitek 2 Compact system combined with K-B disk method was used for drug sensitivity test. The resistance genes were detected by PCR amplification. Plasmid replicon typing was detected by PCR. Plasmid conjugation tests were performed to explore the conjugating transfer of plasmids in the three strains. The homology of the three strains was analyzed by multiple locus sequence typing (MLST) and pulse field gel electrophoresis (PFGE). @*Results@#Drug susceptibility test showed that the three bacteria were resistant to most β-lactam antibiotics except Aztreonam, and resistant to quinolones and SMZ-TMP, but sensitive to aminoglycosides drugs. PCR and sequencing results indicated that the three strains carried bla SHV gene and extended-spectrum beta-lactamase gene (bla SHV , bla TEM and bla CTX-M ). The plasmid replicon type was IncX3. Transfer test of E1 strain was successful. MLST results indicated that all the three strains were ST1642 type. MLST and PFGE results indicated that the bands of the three bacteria were identical. @*Conclusion@#Both NDM-5 carbapenemase and extended-spectrum beta-lactamase were detectable in the three strains of carbapenem-resistant bacteria from neonatal department. MLST and PFGE results suggested that the three strains were from the same clonal source.
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Objective To compare the efficacy of different doses of dexmedetomidine for sedation in patients with Parkinson's disease (PD) undergoing deep brain stimulation (DBS) surgery.Methods Seventy-five patients of both sexes with PD,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,aged 48-72 yr,with body mass index of 18-24 kg/m2,undergoing elective DBS surgery,were divided into 3 groups (n =25 each) using a random number table method:different doses of dexmedetomidine groups (D1-3 groups).Dexmedetomidine was intravenously infused in a loading dose of 0.5 μg/kg at 10 min before local infiltration anesthesia with 0.25 % ropivacaine 10-20 ml,followed by an infusion of 0.3,0.5 and 0.8 μg · kg-1 · h-1 in D1-3 groups,respectively.The infusion of dexmedetomidine was suspended during neurological test,the patients were awakened,and dexmedetomidine was infused at 0.3,0.5 and 0.8 μg · kg-1 · h-1 until the end of the surgery after the test.Bispectral index (BIS) value was recorded immediately before anesthesia (T0),immediately after administration prior to wake-up (T1) and during neurological test (T2).The wake-up time and adverse reactions such as requirement for rescue analgesics,pulse oxygen saturation (SpO2) < 90% and snoring were recorded.Results Compared with group D1,the requirement for rescue analgesics and BIS value at T1,2 were significantly decreased in D2 and D3 groups,and the wake-up time was significantly prolonged,and the incidence of SpO2 < 90% and snoring was increased in group D3 (P<0.05 or 0.01).Compared with group D2,the requirement for rescue analgesics and BIS value at T1 were significantly decreased,the wake-up time was prolonged,and the incidence of SpO2 <90% and snoring was increased in group D3 (P<0.05 or 0.01).Conclusion Dexmedetomidine 0.5 μg · kg-1 · h-1 can produce good sedative efficacy and safety for DBS surgery in patients with PD.
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Objective To analyze the drug resistance phenotype and genetic background of New Delhi metallo-β-lactamase (NDM)-producing bacterial strains in Lishui area of Zhejiang province.Methods The imipenem-resistant Enterobacteriaceae strains were isolated from January 2012 to December 2016 in Lishui Municipal Central Hospital of Zhejiang Province.Mrieux Vitek 2 Compact system was used to identified strains and PCR was used to screen for blaNDMgene.Susceptibility was detected by K-B method and MICs were obtained by Vitek 2 with GN13 cards.Plasmids typing was carried out by DNA sequencing of the replication initiator with the transconjugates as templates.The blaNDMgenetic contexts were detected by PCR and complete genome sequencing.Results A total of 102 strains of carbapenem-resistant enterobacteria (CRE), mainly Escherichia coli and Enterobacter cloacae, were isolated, of which 15 were positive for blaNDMwith a positive detection rate of 14.7%.The resistance rate to β-lactam antibiotics was 100%, and the resistance rates to aztreonam, compound sulfamethoxazole , tobramycin and gentamicin were all >80%;and the resistance rate to quinolones was >50%.Among the 15 NDM-producing strains , 12 strains were positive for Hodge test, and 2 strains of Enterobacter cloacae and 1 strain of Escherichia coli were negative. There were 11 strains of blaNDM-1and 4 strains of blaNDM-1.A variety of plasmid types such as IncX 3, IncFIIγ and IncA/C were detected, and 4 blaNDM-5genes were located in the IncX3 plasmid.Four blaNDMsurrounding gene structures were found, of which 8 blaNDM-1genes were located in ISAb125-hyp-blaNDM-1-bleMBL-TrpF-DsbC-IS26, while blaNDM-5was located in IS3000-IS5-blaNDM-5-bleMBL-TrpF-DsbC-IS26 structure, and the former was reported for the first time in China.Conclusion NDM-producing bacterial strains in Lishui area are prevalent at a low level and have high sensitivity to fosfomycin and polymyxin .The blaNDMgene may have multiple sources, but IncX3 plasmid is still the main source for gene transfer.Some new types of blaNDM-1 gene structure have been also found in this study.
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Objective@#To investigate the distribution of blaKPC gene in Lishui and to analyze the molecular epidemiological characteristics of Klebsiella pneumoniae (K.pneumoniae) blaKPC gene.@*Methods@#From 2010 to 2016, all of the non-repetitive K. pneumoniae carbapenemase (KPC)-producing isolates in Lishui Municipal Central Hospital were collected. They were identified with VITEK 2 Compact system and typed by multilocus sequence typing (MLST). Plasmids were classified based on the DNA sequences of replication initiators. Transposons were detected by PCR. Locations of blaKPC gene were verified through complete sequencing of the plasmids by next-generation sequencing (NGS).@*Results@#A total of 125 strains were collected. K. pneumoniae strains accounted for 88.8% (111) and among them, 103 were ST11 type. IncF plasmids were detected in 48.6% of K. pneumoniae strains and most of them carried mutant Tn1721/Tn4401 chimera (48/54 isolates). Untypable plasmids were discovered in 50.5% of isolated strains and most of them were positive for the wild-type chimera (54/56 isolates). IncF-positive strains isolated during the period of 2011 and 2013 accounted for 94.4%, followed by a dramatic decrease. However, 76.8% of the strains harboring untypable plasmids were isolated from 2014 to 2016 and the number increased year by year.@*Conclusion@#K. pneumoniae of ST11 type was the main cause of blaKPC gene dissemination in Lishui area. Strains carrying the IncF plasmids integrated with the mutant Tn1721/Tn4401 chimera and the untypable plasmids with the wild-type chimera were prevalent before and after 2014, respectively.
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Objective To observe the effect of AS1411-mediated signal transduction and activator of transcription 3 (STAT3) antisense oligonucleotide (ASO) targeting tumor cells.Methods RNA was used as coupling molecules to link the targeting molecules AS1411 and effector molecules ASO.Prediction and analysis of the secondary structure of the pre-synthesis of chimeric molecules by RNA Structure software.Agarose gel electrophoresis was used to test the stability of chimeric molecules in serum and cell lysis solution.Using flow cytometry and confocal fluorescence microscope were used to estimate the internalization of AS1411-mediated STAT3 ASO.Inhibitory effect of ASO on the growth of tumor cells was detected by CCK-8 kit.RT-PCR and Western blot was used to measure the expression levels of tumor related genes.Results STAT3 ASOs mediated by AS1411 can enter tumor cells efficiently to inhibit the transcription and translation of C-myc,Cyclin D1,Bcl-xl and PD-L1 gene,and also can inhibit the growth of Du145 cells.Conclusions AS1411-mediated STAT3 ASO can enter tumor cells and act as anti-tumor medicine.
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Objective To evaluate the efficacy of a type of endotracheal tube of extended with terminal swelling and resistance bending pressure for airway management during mechanical ventilation in children in prone position.Methods Sixty pediatric patients with cerebral palsy of both sexes,aged 3-7 yr,weighing 11-23 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective selective functional posterior rhizotomy,were randomly allocated into 2 groups (n =30 each) using a random number table:endotracheal tube of extended with terminal swelling and resistance bending pressure group (group E) and reinforced tracheal tube group (group C).After induction of anesthesia,the type of endotracheal tube of extended with terminal swelling and resistance bending pressure was inserted in group E,and the reinforced endotracheal tube was inserted in group C.The tidal volume,minute ventilation,dynamic lung compliance,arterial oxygen saturation and end-tidal pressure of carbon dioxide in pressurecontrolled ventilation mode were recorded in the two groups.Tube shedding caused by using of the extended tube was recorded during operation.Blood samples were collected from the femoral artery for blood gas analysis.Results Compared with group C,the tidal volume and minute ventilation were significantly increased (P<0.05),and no significant change was found in dynamic lung compliance,arterial oxygen saturation and end-tidal pressure of carbon dioxide in group E (P>0.05).Compared with the value before anesthesia,no significant change was found in pH value,arterial oxygen partial pressure and partial pressure of arterial carbon dioxide at the end of operation in the two groups (P>0.05).In group C,the extended tube wasused in 3 patients,and among the 3 cases,one patient developed tube shedding which was treated immediately.Conclusion The type of endotracheal tube of extended with terminal swelling and resistance bending pressure provides better efficacy than the reinforced endotracheal tube when used for airway management during mechanical ventilation in children in prone position.
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Objective To evaluate the accuracy of nostril width method in intubation model choose in children with cerebral pal‐sy FSPR surgery .Methods Three hundred and fifty six cases of cerebral palsy FSPR surgery were selected ,the endotracheal intu‐bation were selected by nose width method and modified weight method ,and then we made a comparison between the two methods . Kappa consistency test and rank correlation analysis were both used to analyze the consistency .Results With Kappa=0 .81 ,rank correlation coefficient 0 .905 ,we could know that there was high degree of consistency between the two methods (P<0 .01) ,of which 298 cases (83 .7% ) children selected the same model intubation strengthen model in different method ,58 cases (16 .3% ) pe‐diatric intubation choice of two methods differ by one model .Conclusion Nostril width method was simple and reliable to select in‐tubation in pediatric surgery to strengthen FSPR model .
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BACKGROUND:Poly(3-hydroxybutyrate) is approved as its excel ent biocompatibility, biodegradability and piezoelectric properties, but there are also some deficiencies, such as high breakability and poor hydrophilicity. METHODS:Poly(3-hydroxybutyrate) was mixed with different mass percentages of nanohydroxyapatite (0, 10%, 20%and 30%) to prepare new composite fibrous scaffolds through electrospinning process. The microstructure, group composition, crystal ine phase distribution, thermal properties and surface wettability of the scaffolds were detected. RESULTS AND CONCLUSION:Under the scanning electron microscope, with the increase of nano-hydroxyapatite content, more and more nano-hydroxyapatite particles were distributed evenly on the composite fiber surface;the fiber surface was basical y covered with nano-hydroxyapatite particles at the content of 30%, and the roughness of the fiber surface also increased. Results from differential scanning calorimetry and X-ray diffraction showed that the nano-hydroxyapatite reduced the crystal inity of poly(3-hydroxybutyrate) and the crystal tacticity, and this phenomenon became more evident with the increase of nano-hydroxyapatite content. Additional y, the higher the content of nano-hydroxyapatite content, the lower the contact angle and the higher the hydrophily. These findings indicate that the nano-hydroxyapatite/poly(3-hydroxybutyrate) ultrafine-fibrous scaffold using electrospinning technology can effectively improve the surface wettability and crystal inity of the material as wel as the material hydrophily and brittleness, and the higher the content of nano-hydroxyapatite, the more obvious the effect.
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Objective To evaluate the effect of cerebral palsy factor on the sensitivity of postopera?tive pain in the pediatric patients. Methods Twenty?five pediatric patients with cerebral palsy of both se?xes, of American Society of Anesthesiologists physical statusⅠorⅡ, aged 3-7 yr, weighing 11-25 kg, scheduled for elective lower abdominal or lower extremity surgery, served as cerebral palsy group ( group P). Another 25 pediatric patients without cerebral palsy of both sexes, of American Society of Anesthesiol?ogists physical status Ⅰ orⅡ, aged 3-7 yr, weighing 11-25 kg, served as control group ( group C) . At 2 h after surgery, pain was evaluated by using CRIES ( crying, requires O2 saturation, increased vital sign, expression and sleeplessness) . Peripheral venous blood samples were collected before surgery, and at 2 and 24 h after surgery, and the concentrations of plasmaβ?endorphin were measured by radio?immunity method, and the concentration of plasma catecholamine ( adrenaline) was determined by high performance liquid chromatography. Results Compared with the value before surgery, the plasma concentrations of β?endorphin were significantly decreased, and the concentrations of plasma catecholamine were increased after surgery in the two groups (P < 0?01). Compared with group C, the CRIES score was significantly in?creased after surgery, the concentration of plasmaβ?endorphin was decreased before and after surgery, and the concentration of plasma catecholamine was increased after surgery in group P ( P< 0?05 or 0?01) . Con?clusion The sensitivity of postoperative pain is increased in the pediatric patients with cerebral palsy.
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Objective To study the capacity of dentifrice containing hydroxyapatite (HA) particles with various grain sizes on the sorption of Cu (Ⅱ) ion from aqueous solution. Methods Three groups of dentifrice suspensions containing 3% HA particles with 60 nm grain size, 3% HA particles with 12 μm grain size, and blank dentifrice were prepared with serial concentrations. Copper ion solutions with initial concentration of 4.0 mg/L were mixed with dentifrice suspensions respectively and then been kept still for 24 hours. The residual copper ion concentration of the supernatant was measured by inductively coupled plasma emission spectrometer and the sorption rate of copper ion was calculated. Results The residual copper ion concentration of the supernatant decreased with increase of the concentration of dentifrice suspensions. The sorption rate of copper ion of dentifrice containing 60 nm HA was highest among the three groups. Conclusion Nano HA should be added into dentifrice to enhance the sorption capacity of copper ion from aqueous solution.
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Objective To histomorphometricly assess changes occurred in the alveolar ridge following different methods of socket preservation and to compare them against natural healing without interventions. Methods The second、 third and fourth mandibular premolars were extracted from six beagles. Six extraction sites in each dog were randomly assigned to three treatments as follows:natural healing (T1), Bio-Oss Collagen (T2) and immediate implant with Bio-Oss (T3). Six month after surgery, the dogs were euthanized and tissue samples were sectioned, fixed and mounted, then were stained with toluidine blue. The histologic studies and morphological measurements were performed by using an optical microscope and a digital image software. Results Reabsorption in the buccal aspect of the alveolar crest of ridge was showed in all groups. With respect to the mean vertical bone loss of the buccal bone plate, T3 is lower than T1 and T2(P<0.001 ), while no significant differences were observed between T1 and T2. With regard to horizontal dimension of the alveolar process , a statistical significance could be found at 3mm and 4mm below the crest of ridge in group T1 and T3(P=0.017, P=0.042), while no statistical differences were found between other groups. Conclusions Both techniques of alveolar ridge preservation were not able to completely preserve the original bone volume after tooth extraction. Immediate implant placement in combination with Bio-Oss seems to have the potential to limit the reabsorption of the alveolar process efficiently , but the bone preserving effect of Bio-Oss Collagen is undesirable.
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Objective To investigate the effect of dexmedetomidine on emergence agitation (EA) following sevoflurane anesthesia in children with cerebral palsy.Methods Fifty ASA physical status Ⅰ or Ⅱ children with cerebral palsy of both sexes,aged 2-12 yr,scheduled for elective muscle strength muscle tension adjustment method,were randomly allocated into 2 groups (n =25 each):control group (group C) and dexmedetomidine group (group D).Anesthesia was induced with iv injection of sufentanil 0.3 μg/kg,cisatracurium 0.15 mg/kg,and propofol 1.5 mg/kg.The patients were tracheal intubated and mechanically ventilated.Dexmedetomidine 0.5 μg/kg was intravenously infused over 15 min after induction of anesthesia in group D and the equal volume of normal saline was given in group C.The operation was begun at the end of administration.Anesthesia was maintained with inhalation of 2%-4% sevoflurane and the concentration was adjusted to achieve a target BIS value of 45-60.The heart rate,systolic blood pressure and diastolic blood pressure were recorded before infusion of dexmedetomidine (T1) and at skin incision (T2).The extubation time,emergence time and consumption of sevoflurane were also recorded.The end-tidal sevoflurane concentration was recorded at T1,T2,and the end of operation (T3).The adverse cardiovascular events and occurrence of EA were also recorded.The degree of EA was assessed using the Pediatric Anesthesia Emergence Delirium (PAED) scale.The peripheral venous blood samples were collected at T1,T2,T3 and extubation (T4) for determination of the blood glucose level and serum cortisol concentration.Compared with group C,the emergence time and extubation time were significantly shortened,and the consumption of sevoflurane,end-tidal sevoflurane concentrations at T2 and T3,PAED scores,incidence of agitation,and the blood glucose level and serum cortisol concentration at T4 were significantly decreased in group D (P <0.05 or 0.01).There was no significant difference in the incidence of bradycardia or heart rate,systolic blood pressure and diastolic blood pressure at each time point between the two groups (P > 0.05).Conclusion Dexmedetomidine can decrease the development and degree of EA following sevoflurane anesthesia in children with cerebral palsy.
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There are two ways to place dental implants in the maxillary posterior without sufficient bone available, one is the maxillary sinus floor bone augmentation technique, the other is to evade maxillary sinus. The maxillary sinus floor bone augmentation technique may result in a series of complication, and reduce obedience of patients for implant surgery. According to principles of the best bone utilization, the evaded maxillary sinus implant technique is more simple, more economic, more predictable, and more shortage of time, compared with the maxillary sinus floor bone augmentation technique, and has certain technological advantages in some special cases.
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Humans , Dental Implantation, Endosseous , Dental Implants , Maxilla , Maxillary Sinus , Risk Factors , Sinus Floor AugmentationABSTRACT
Objective To investigate the clinical results of immediate loading of a single anterior implant. Methods 26 implants were placed into anterior sites. Provisional prosthesis were placed at the time of fixture placement. It did not occlude in any pathway. Restoration was for esthetic only, not for function. Results No fixture was lost since the immediate restoration was delivered, the minimum being 12 months and the maximum 72 months. Conclusion Clinical studies on the placement of immediate restoration in anterior implant of maxilla have revealed predictable results.
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OBJECTIVE To detect genes sul1 and sul2 in Stenotrophomonas maltophilia,and their relationship to drug resistance to trimethoprim-sulfamethoxazole(SXT).METHODS K-B was carried out to detect the drug resistance to SXT of S.maltophilia;minimal inhibitory concentration(MIC) was measured with micro broth dilution method.Genes sul1 and sul2 were amplified by PCR.RESULTS Eight isolates(7.8%) showed resistance to trimethoprim-sulfamethoxazole,sul1 Was positive in four isolates in which one contained sul2 also.Four isolates showed high MIC to SXT.CONCLUSIONS Genes sul1 and sul2 of S.maltophilia are associated with the high drug resistance to SXT.
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Objective: To observe the role of Qudu medicinal granules (祛毒冲剂) on enterogenous endotoxemia . Methods: Sixty-one cases with enterogenous endotoxemia were randomly divided into two groups:Qudu medicinal granule group (n=30) that was treated with Qudu medicinal granules combined with western medicine, and smecta group which was treated with smecta and western medicine (n=31). Changes of symptoms and signs were observed before treatment and 1, 3, 7 days after treatment. Blood samples were collected in the morning to measure the white blood cell (WBC), plasma lipopolysaccharide (LPS) and tumor necrosis factor (TNF) levels. Results: Recovery speed of WBC count in Qudu medicinal granule group was faster than that of the smecta group, there was significant difference on the third day after treatment (P
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Objective To establish attenuated S. typhimurium expressing Helicobacter pylori (H.pylori) urease subunit B (UreB) and determine whether it could be used as oral vaccine against H.pylori Methods H.pylori (SS1 strain) UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector PTc 01 after sequencing, then transformed into attenuated S.typhimurium SL3261 to acquire SL3261/PTc 01 UreB. The expression of H.pylori UreB in SL3261 was detected by Western blot. Twelve weeks after oral immunization of mice, antibody responses were evaluated using serum and intestinal fluid by ELISA. In vitro stability of PTc 01 UreB plasmid in S. typhimurium SL3261 was confirmed by growing in Luria Bertani medium to 60 generations. Results The UreB gene fragment amplified by PCR was consistent with the sequence of the H.pylori UreB by sequence analysis. Enzyme digestion revealed that the correct PTc 01 UreB was obtained. Western blot showed that 61kD protein was expressed in SL3261/PTc 01 UreB which could be recognized by anti H.pylori UreB antiserum. Anti UreB IgA antibodies in mouse intestinal fluid and IgG antibodies in serum were determined by ELISA. After 60 generations of continuous culture, the recombinant plasmid PTc 01 UreB was stable in SL3261 and had no obvious toxicity. Conclusion The attenuated Salmonella typhimurium expressing H.pylori UreB may be used as oral vaccine against H.pylori infection.
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Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1∶ 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1∶ 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.