Analysis of AmpC β-lactamase Gene in Pseudomonas aeruginosa

The gene and the amino acid sequence of the structural and regulatory region of the Pseudomonas aeruginosa with different resistance patterns were analyzed. Six strains with different resistance patterns were selected and the AmpC β-lactamase was identified. The objectivegene fragment was amplified by colonies PCR. The sequences of the PCR-products were analyzed. The DNA sequence of the structural gene ampC and the regulatory genes ampR, ampD and ampE was detected. The 6 strains and the wild-type Pseudomonas aeruginosa are highly homogeneous in structural and regulatory region. Some new mutant points were found.     

Transconjugation and genotyping of the plasmid-mediated AmpC β-lactamase and extended-spectrum β-lactamase genes in Klebsiella pneumoniae

Bsckgroud AmpC β-lactamases and extended-spectrum β-lactamases （ESBLs） are becoming predominant causes of resistance to third and forth-generation cephalosporins in Klebsiella pneumoniae （K. pneumoniae）. It is very difficult to treat infectious diseases caused by multidrug-resistant K. pneumoniae. The purpose of the present study was to investigate transconjugation and characteristics of β-lactamase genes in K. pneumoniae producing AmpC β-lactamases and ESBLs.
Methods AmpC β-lactamases were detected by three-dimension test and ESBLs by disc confirmatory test. Minimum inhibitory concentrations （MICs） were determined by agar dilution. Transfer of resistance to EC600 （Rifr） was attempted by conjugation in broth and screened on agar containing cefotaxime （2 μg/ml） plus rifampin （1024 μg/ml）. The genes encoding AmpC or ESBLs and their transconjugants were detected by PCR and verified by DNA sequencing.
Results The resistant rates to ampicillin and piperacillin were 100% in 18 isolates of K. pneumoniae. However, imipenem was still of great bactericidal activity on K. pneumoniae, and its MIC50 was 0.5 μg/mL. Eleven β-lactamase genes, including TEM-1, TEM-11, SHV-13, SHV-28, CTX-M-9, CTX-M-22, CTX-M-55, OXA-1, LEN, OKP-6 and DHA-1, were found from 18 isolates. And at least one β-lactamase gene occurred in each isolate. To our surprise, there were six β-lactamase genes in the CZ04 strain. Among 18 isolates of K. pneumoniae, the partial resistant genes in 8 isolates were conjugated successfully, which had 100% homological sequence with donors by sequence analysis. Compared with donors, 8 transconjugants had attained resistance to most β-lactams, including ampicillin, piperacillin, cefoxitin, cefotaxime and aztreonam, or even amikacin and gentamicin.
Conclusions R plasmids can be easily transferred between the resistant and sensitive negative bacilli. It is very difficult to block and prevent the spread of antimicrobial resistance. So more attention should be paid to reducing the frequency, times and dosage of antimicrobials, especially third or fourth cephalosporins.     

One-year outcomes of percutaneous renal denervation for thetreatment of resistant hypertension： the first Chinese experience

Background As a novel device-based approach targeting the renal sympathetic nerves, percutaneous renal denervation（RDN） has been shown to be effective and safe for reducing blood pressure. However, while considerable data on RDNhave been obtained from Western populations, there is limited findings from East Asian populations. The purpose of thisstudy was to evaluate one-year outcomes of RDN for the treatment of resistant hypertension in Chinese patients.Methods Between February and August 2012, 14 patients （mean age 39＋8 years, 10 males） with resistant hypertensionunderwent successful RDN at the Fuwai Hospital. All 14 patients were followed up at 1, 3, 6 and 12 months post-procedure. Blood pressure, use of antihypertensive agents, renal function, and complications were investigated.Results Baseline values included mean office blood pressure of 164/103＋14/10 mmHg, mean 3.9＋0.6 anti-hypertensiveagents, and an estimated glomerular filtration rate of （79＋19） ml·min-1·1.73 m-2. Office blood pressure after the procedurewas reduced by -14/-10, -17/-11, -21/-12, and -24/-14 mmHg at 1, 3, 6, and 12 months respectively, and thereduction of the number of antihypertensive agents at the above corresponding time points was -1.3, -1.5, -1.7 and -1.8respectively （all P 〈0.001）. The mean reduction of 24-hour ambulatory blood pressure was similar to the reduction of officeblood pressure at the four corresponding time points. Renal function did not significantly change at any time point （all P〉0.05）. No clinical complications were observed at 12-month follow-up.Conclusion This study showed that RDN seems to be effective in reducing blood pressure of Chinese patients withresistant hypertension, with minimal adverse events at 12-month follow-up.     

Analysis on the production of extended-spectrum β-Lactamases and drug-resistance of 906 strains of Escherichia coli

目的 了解大肠埃希菌产超广谱β-内酰胺酶(ESBLs)及其对常用抗菌药物的耐药情况,为临床抗感染治疗提供用药依据.方法 采用头孢他啶与头孢他啶/克拉维酸、头孢噻肟与头孢噻肟/克拉维酸的双纸片确证试验检测ESBLs,采用纸片扩散法(K-B法)检测大肠埃希菌对常用抗菌药物的耐药性.结果 2007年、2008年、2009年、2010年产ESBLs大肠埃希菌的检出率分别为38.65％、42.86％、43.28％和48.84％.大肠埃希菌对常用抗菌药物的耐药率以头孢哌酮/舒巴坦、亚胺培南、哌拉西林/也唑巴坦、阿莫西林/克拉维酸、头孢西丁、阿米卡星较低,其余抗菌药物的耐药率大多在50％以上,对氨苄西林和哌拉西林的耐药率甚至超过80％.结论 大肠埃希菌的耐药形势严峻,应加强其耐药性监测,合理使用抗菌药物,严格控制耐药菌的产生和医院感染暴发流行.     

Studies on Extraction Process of Volatile Oil from Dajianzhong Granules and Preparation Technology of β-cyclodextrin Inclusion Compound

目的 研究大建中颗粒中挥发油提取工艺及β-环糊精包合物制备工艺. 方法 以提取率为评价指标优化提取工艺,以β-环糊精包合物的收率和挥发油包合率为评价指标,经正交试验法考察包合物制备工艺. 结果 干姜、花椒挥发油的最佳提取工艺为6倍量水浸泡2h,水蒸气蒸馏6h;包合物最佳制备工艺为挥发油与β-环糊精之比为1:6 (mL:g),6倍加水量,包合温度为50℃,超声包合60 min,挥发油包合率为92.1％,包合物收率为89.7％. 结论 优选工艺简单,适合工业化生产.     

Genetic environment of β-lactamase genes of extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates from patients with lower respiratory tract infection in China

Background Extended-spectrum β-lactamase （ESBL）-producing Klebsiella pneumoniae （K.pneumoniae） is one of the most popular pathogens that cause refractory respiratory tract infection.The genetic environment,including insertion sequences and the types of promoter,plays a key role in exploration of the mechanism of prevalence and dismission of the ESBL-producing K.pneumoniae isolates.The aim of the investigation was to target analysis the genetic environment and promoter sequences of blaCTX-M,blaSHV and blaTEM,the most popular β-lactamase genes harbored by ESBL-producing K.pneumoniae isolates.Methods From February 2010 to July 2011,158 of 416 K.pneumoniae isolates producing ESBL from patients with lower respiratory tract infection were collected from seven tertiary hospitals from Beijing,Anhui,Fujian,Liaoning,Hebei and Inner Mongolia Autonomous Region in China.The genetic environment including promoters of 10 types of blaCTX-M,18 types of blaSHVand 2 types of blaTEM were analyzed by amplification and direct sequencing with various sets of PCR primers.Results ISEcp1 was located upstream of the 5＇ end of the blaCTX-M gene in 130 （97.0％） out of 134 K.pneumoniae isolates harboring blaCTX-M and provided a conserved promoter to blaCTX-M.A non-coding sequence preceded by kdpC and recF was identified in all of the blaSHV genes except blaSHV-12 and blaSHV-2a.IS26 was also found upstream of 1 blaCTX-M-15,10 blaSHV-1 strains,4 blaTEM-1 and all of the blaSHV-2,blaSHV-2a,blaSHV-5 and blaSHV-12.Eighty-seven of 91 strains harboring blaTEM-1 carried a copy of Tn2 and IS26-blaTEM-1 fragments were also detected in 4 strains.With respect to K.pneumoniae,the genetic environment of blaCTX-M-38,blaSHV-142 and blaTEM-135 were firstly elaborated,and four kinds of novel genetic environment of blaCTX-M-3,blaCTX-M-15 and blaTEM-1 have been detected as well.Conclusions Perfective implementation of the genetic environment information of β-lactamase gene needs to be further explored and supplemented.ISEcp1 and IS26 elements ar     

Clinical value of monitoring of serum β-HCG and progesterone levels in patients with abnormal pregnancy

目的 通过监测血清β-HCG与孕酮水平,探讨其在诊断早期异常妊娠的临床价值.方法 将我院收治的318例早期异常妊娠患者按照妊娠结局类型分为先兆流产组136例、难免流产组104例和异位妊娠组78例,并对三组血清β-HCG与孕酮水平进行监测.结果 当日检测异位妊娠组β-HCG值及孕酮均最低;48h后检测异位妊娠组血清β-HCG值变化很小( P＞0.05),而先兆流产组血清β-HCG值上升或难免流产组β-HCG值下降变化显著(P＜0.05).结论 血清β-HCG与孕酮水平的联合监测对早期异常妊娠的诊断和治疗有重要的临床意义.     

Gene transfer of a β2-adrenergic receptor kinase inhibitor up-regulates the level of β2-adrenergic receptor and cAMP in the asthmatic murine lung

Objective: To investigate the effects of gene transfer of a β-adrenergic receptor(β-AR) kinase inhibitor(β ARKct)on pulmonary β2-adrenergic receptor and cAMP following β2-AR agonist treatment in asthmatic mice, and to analyze the relationship between the routes of gene delivery and the changes of β2AR and cAMP. Methods: BALB/c mice were sensitized and challenged by ovalbumin to establish the asthmatic model treated with βAR agonist ( salbutamol injected intramuscularly). The plasmid with the expression of βARKct was constructed and βARKct gene transfer was performed through intravenous injection or intratracheal instillation in asthmatic mice.The gene expression was measured with Western blot analysis, and the changes of pulmonary β-AR and cAMP evaluated by Radioimmunoassay. Results: The expression of tranfered βARKct gene was detectable in lungs and it was expressed more in the lungs of the mice receiving intratracheally plasmid than those receiving intravenously. The levels of βAR and cAMP were upregulated after using plasmid-βARKct to the asthmatic mice treated with β AR agonist. Conclusion: Our results indicated that there were down-regulation of βAR and cAMP in asthmatic mice treated with βAR agonist. Gene transfer of βARKct could inhibit the extent of the down-regulation of βAR and cAMP. The route of gene delivery could also affect the degree of up-regulation of βAR and cAMP. Gene transfer βARKct may provide a novel approach to the therapeutic strategy for asthma.     

Fetal Liver Transplantation in the Treatment of β-Thalassemia Major:Report of Two Cases

Two cases of β-thalassemia major treated with fetal liver transplanta-tion(FLT)were reported.A conditioning regimen of total body irradiation pluscyclophosphamide was given,followed by the infusion of suspensions of fetal livercells.The number of nucleated cells(procared from 3 to 4 fetal livers)infused tothe 2 cases was 17.5 and 15.5×10~8/kg respectively.Transient hemopoieticdepression was observed after FLT,and normal hemopoiesis was recoveredaround the 30th day.Karyotypic analysis revealed that the donor typechromosomes was founed in the peripheral blood of both cases from the 20th dayto 123rd day after FLT.Skin biopsy of cass 1 revealed changes of mild GVHD,indicating engrafment of fetal hemopoietic cells and formation of chimeras in thisperiod.Autologous hemopoiesis was reconstituted in both cases two months afterFLT.Blood transfusion was not necessary within the first 5 months after FLT,and in case 1 transfusion was given 3 times only within 21 months after FLT.     

Clinical efficacy and safety of Gubitong Recipe (骨痹通方) in treating osteoarthritis of knee joint

Objective:To observe the therapeutic efficacy and safety of Gubitong Recipe(骨痹通方,GBT) in treating osteoarthritis(OA) of knee joint.Methods:Ninety patients with knee osteoarthritis were equally assigned, according to a randomizing digital table,to the treatment group and the control group.The treatment group was treated with GBT Decoction one dose every day and the control group with glucosamine sulfate 500 mg thrice a day,respectively,for eight successive weeks.Besides,diclofenac sodium could be given as...     

Differential expression of βig-h3 in human hepatoma cell lines

Objective：To observe the differential expression of TGF-β-induced gene human clone 3 （βig- h3） in human hepatoma cell lines. Methods：Human hepatoma cells HHCC, 7721, T7721 constructed by stably transfectiug HAb18G/CD147 cDNA into 7721 and normal human liver cell QZG were cultured as previously. RT-PCR and western blot were used to investigate the differential expression of βig-h3 in human hepatoma cell lines and normal liver cell. Results ：The results of RT-PCR suggested that the expression of βig-h3 mRNA in human hepatoma cells was higher than that in normal human liver cell QZG（P〈0.01）, and its expression level in human hepatoma cells in turn was HHCC〉T7721〉7721. Moreover, the similar results of βig-h3 protein expression were testified by western blot. Conclusion：This study demonstrates that the expression of βig-h3 in hepatoma cell lines is higher than that in normal liver cell QZG, which provides a sound basis for exploring the function of βig-h3 in processes of adhesion and metastasis of human hepatoma cells.     

Expression of βig- h3 in keratoconus and normal cornea

Objectives To observe the expression of βig- h3 in normal cornea and keratoconus and to elucidate the role of extracellular matrix in keratoconus.Methods In situ hybridization was used to detect the expression of βig- h3 in the cornea.The cDNA library was screened with human βig- h3 cDNA probe to locate βig- h3 mRNA in cells.Results Expression of βig- h3 was found mainly in the stroma of the normal cornea and keratoconus, but decrease depending on the degree of keratopathy.In some serious cases, no expression signal was detected. The strongest expression was seen at the border of the normal region and keratoconus.Conclusions βig- h3,the structural component of the extracellular matrix, can affect cell adhensiveness in the development of corneal fibrous interstitial organization.During the development of keratoconus, decreasing levels of βig- h3 cause the diminution of corneal steadiness, which is related to formation of keratoconus     

Association of Serum Dkk-1 Levels with β-catenin in Patients with Postmenopausal Osteoporosis

Wnt signaling plays an important role in the bone development and remodeling. The Wnt antagonist Dkk-1 is a potent inhibitor of bone formation. The aims of this study were firstly to compare the serum Dkk-1 levels in postmenopausal osteoporosis patients with age-matched healthy controls, and secondly, to assess the possible relationship between Dkk-1 and β-catenin, sclerostin, or bone turnover markers [CTX, PINP, N-MID-OT and 25(OH)D] in the setting of postmenopausal osteoporosis. A total of 350 patients with postmenopausal osteoporosis and 150 age-matched healthy controls were enrolled, and the serum levels of Dkk-1, β-catenin, sclerostin, OPG, and RANKL were detected by ELISA, and bone turnover markers [CTX, PINP, N-MID-OT and 25(OH)D] were measured by Roche electrochemiluminescence system in two groups. Serum Dkk-1 levels were significantly higher in postmenopausal osteoporosis group than in control group(P<0.001). Univariate analyses revealed that serum Dkk-1 levels were weakly negatively correlated to β-catenin(r=–0.161, P=0.003) and OPG(r=–0.106, P=0.047), while multiple regression analysis showed a negative correlation between serum Dkk-1 levels with β-catenin(β=–0.165, P=0.009) and BMD(β=–0.139, P=0.027), and a positive correlation between serum Dkk-1 levels and CTX(β=0.122, P=0.040) in postmenopausal osteoporosis group. No similar correlations ware observed in control group. The results provided evidence for the role of Dkk-1 in bone metabolism and demonstrated the link of Dkk-1 and Wnt/β-catenin in some ways.     

Effects of plasma β-endorphin on hemorrhagic shock in acute hypoxic rats

Forty rots were randomized into 2 groups and naloxone or saline were injected to therats after they were inflicted with hemorrhagic shock at sea level and at a simulated altitude of4000m respectively to observe the effects of naloxone on left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVDP),the maximal changing rate of LVSP(dp/dt max),heartrate(HR),and survival time of the animals.Plasma β-endorphin(β-EP)was determined beforeand after hemorrhage to observe the relationship between β-EP and hemorrhagic shock.It wasfound that the circulatory parameters of hemorrhagic shock changed more markedly at high alti-tude than at sea level,naloxone could restore these parameters and prolong the survival time inboth the animals of the sea level and high altitude groups,and plasma β-EP level was elevatedafter hemorrhage especially in those animals at high altitude.These findings indicate:(1)Hemorrhagic shock at high altitude is usually accompanied with severe clinical manifestations,rapid progression,and high mortality.(2)β-EP seems to participate in the pathologicalmanifestafions of hemorrhagic shock at high altitude,and its depressive action on myocardialcontraction may be one of the factors inducing hemorrhagic shock.(3)Naloxone possesses defi-nite property to comet hemorrhagic shock at high altitude.     

Changes of Immunoreactive β-Endorphin in Plasma, Pituitary and Hypothalamus of Rats during Oxygen-Induced Convulsions

We observed for the first time the differences of immunoreactive β-endorphin(IR -β- EP) content in plasma, pituitary and hypothalamus of rats under various conditionsusing radioimmunoassay (RIA) and the effects of naloxone and β - endorphin (β- EP) antiserumon initial time of convulsions (ITC), severity of convulsions(SOC) and mortality on surface(MOS) of rats to hyperbaric oxygen(HBO). The results suggest thatβ- EP may partici-pate in the course of oxygen - induced convulsions and be one of endogenous convulsion - causingagents.     

Non-Fusion and Fusion Expression of β-Galactosidase from Lactobacillus bulgaricus in Lactococcus lactis

Objective To construct four recombinant Lactococcus lactis strains exhibiting high β-galactosidase activity in fusion or non-fusion ways, and to study the influence factors for their protein expression and secretion. Methods The gene fragments encoding β-galactosidase from two strains of Loctobacillus bulgaricus, wch9901 isolated from yogurt and 1.1480 purchased from the Chinese Academy of Sciences, were amplified and inserted into lactococcal expression vector pMG36e. For fusion expression, the open reading frame of the β-galactosidase gene was amplified, while for non-fusion expression, the open reading frame of the β-galactosidase gene was amplified with its native Shine-Dalgarno sequence upstream. The start codon of the β-galactosidase gene partially overlapped with the stop codon of vector origin open reading frame. Then, the recombinant plasmids were transformed into Escherichia coli DH5α and Lactococcus lactis subsp, lactis MG1363 and confirmed by determining β-galactosidase activities. Results The non-fusion expression plasmids showed a significantly higher β-galactosidase activity in transformed strains than the fusion expression plasmids. The highest enzyme activity was observed in Lactococcus lactis transformed with the non-fusion expression plasmids which were inserted into the β-galactosidase gene from Lactobacillus bulgaricus wch9901. The β-galactosidase activity was 2.75 times as high as that of the native counterpart. In addition, β-galactosidase expressed by recombinant plasmids in Lactococcus lactis could be secreted into the culture medium. The highest secretion rate （27.1%） was observed when the culture medium contained 20 g/L of lactose. Conclusion Different properties of the native bacteria may have some effects on the protein expression of recombinant plasmids. Non-fusion expression shows a higher enzyme activity in host bacteria. There may be a host-related weak secretion signal peptide gene within the structure gene of Lb. bulgaricus β-galactosidase, and its translation pr     

Expression of claudin-3 and β-catenin in borderline ovarian tumors and its clinicopathological significance

目的 研究claudin-3和β-连环素(β-catenin)蛋白在卵巢交界性肿瘤(borderline ovarian tumors,BOTs)组织中的表达及临床病理意义.方法 采用免疫组织化学SP法检测claudin-3和β-catenin蛋白在69例BOTs组织中的表达,并与其在17例正常卵巢组织、31例卵巢上皮性良性肿瘤、39例卵巢上皮性恶性肿瘤组织中的表达进行比较.结果 BOTs组织中claudin-3蛋白的阳性表达率(59.4％,41/69)低于卵巢癌组织中的阳性表达率(89.7％,35/39),而高于良性肿瘤及正常卵巢组织中的表达率(22.6％,7/31;5.9％,1/17),差异均有统计学意义(P＜0.01);正常卵巢组织与良性肿瘤相比,差异无统计学意义(P＞0.05).BOTs组织中β-catenin蛋白的阳性表达率(52.2％,36/69)低于卵巢癌组织中的阳性表达率(84.7％,33/39),而高于良性肿瘤及正常卵巢组织中的表达率(29.0％,9/31;11.8％,2/17),差异均有统计学意义(P＜0.01或P＜0.05);而正常卵巢组织与良性肿瘤相比,差异无统计学意义(P＞0.05).Claudin-3和β-catenin蛋白的表达与BOTs患者的临床病理分期以及腹腔有无种植相关(P＜0.05),而与组织学类型和患者年龄无关(P＞0.05).Claudin-3与β-catenin在BOTs组织中的表达呈正相关(r=0.439,P=0.000).结论 Claudin-3与β-catenin在BOTs的发生、发展中可能起着协同作用,对其进行联合检测有助于BOTs的诊断和治疗.     

Expression of the Gene Encoding the Tetraploid of Carboxyl-terminal Peptide of β-hCG Containing Thirty-seven Amino Acid Residues in E. coli

Human chorionic conadotropin (hCG) is a glycoprotein secreted by the trophoblast during early pregnancy and plays an important role in the maintenance ofearly pregnancy"'. Immuno--contraception directed against hCG is an effective meanso f fertility regulation. The worldwide interests have centred on developing immunocontraceptive vaccines against hCG, such as the prototype anti--hCG vaccine, advanced prototype anti--hCG vaccine, optimized anti--hCG vaccine and oral anti--hCGvaccine. Eve…