Objective To purify AmpC beta-lactamase from Enterobacter cloacae and study its physical and chemical characteristics.Methods The bacteria were fragmentized by ultrasonication and AmpC beta-lactamase was purified by ion-exchange.The properties of the purified enzyme,such as molecular weight and isoelectric point were determined by electrophoresis.Results The purified AmpC beta-lactamase was obtained and its specificity to substrate was sufficient for classification of class C cephalosporinase.Conclusion AmpC can be successfully purified from Enterobacter cloacae and is effective for application.

AIM: To establish a fluorogenic quantitative polymerase chain reaction (FQ-PCR) method for the routine examination of c-erbB-2 gene expression in breast cancer. METHODS: The c-erbB-2 standard gene was obtained by in vitro amplification of cloned c-erbB-2 fragment in plasmid PGEM-T easy vector. FQ-PCR product was detected by using a 7700 ABI PRISM sequence detector system and c-erbB-2 standard curve was obtained to quantity c-erbB-2 in unknown samples. RESULTS: “S” kinetics curve of FQ-PCR amplification was generated by relating the fluorescence signal intensity (△Rn) to the cycle number. The standard curve of c-erbB-2 was constructed by the linear relationship between the cycle threshold (ct) and the log of starting copy number. The high correlation (0.999) revealed the reliability of FQ-PCR. CONCLUSION: The FQ-PCR is a rapid, sensitive, reliable method for quantity of c-erbB-2 gene expression.

OBJECTIVE To investigate the disinfectant-resistant gene qacE△1-sul1 in Pseudomonas aeruginosa isolated from burned patients. METHODS GNS-448 and K-B tests were performed to detect the susceptibility of 19 kinds of antimicrobial agents against these strains. Genotype was analyzed by polymerase chain reaction(PCR) and verified by DNA sequencing. RESULTS The 32 strains isolated were all resistant to ampicillin,cefuroxime,cefoxitin,SMZ/TPM. The sensitive rates to amikacin and gentamicin were 68.0%,and 46.9%,respectively,the resistant rates to imipenem and meropenem were 68.8% and 59.4%,respectively. The positive rate of gene qacE△1-sul1 was 50.0%. CONCLUSIONS The resistance of P. aeruginosa isolated from burned patients is a serious issue.There is high positive percentage of qacE△1-sul1 gene in P. aeruginosa isolated from burned patients.

[Objective] To observe the effect of Allergic Rhinitis Oral Liquid (AROL) on perennial allergic rhinitis (PAR) and to explore its therapeutic mechanism. AROL, composed of modified Guizhi Tang and Yupingfeng San, has the actions of strengthening spleen and lung, replenishing Qi and consolidating superficies and dispelling pathogens and dredging orifices. [Methods] One hundred and twenty-nine cases of confirmed PAR were randomized into Group A (n=43), Group B (n=43) and Group C (n=43). Group A was treated with AROL, Group B with Biyankang Tablets and Group C with normal saline. Three groups were treated for 4 weeks as one treatment course, and another treatment course was carried out one week after interval. Therapeutic effect were observed after treatment and serum levels of interleukin-4 (IL-4) and specific IgE antibody were detected. [Results] The total effective rate was 93.02% in Group A (P

OBJECTIVE To investigate the 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes in Pseudomonas aeruginosa isolated from burned patients. METHODS GNS-448 and K-B tests were performed to detect the susceptibility to 19 kinds of antimicrobial agents against these strains. 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes were amplified by polymerase chain reaction (PCR) and verified by DNA sequencing. RESULTS The 32 isolated strains were all resistant to ampicillin,cefuroxime,cefoxitin,SMZ-TMP,The sensitive rates to amikacin and gentamicin were 68% and 46.9%,respectively. The resistant rates to imipenem and meropenem were 68.8% and 59.4%,respectively. The 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes including aac(6')-Ⅰb,aac(6')-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and rmtB were found and positive rates were 9.4%,3.1%,28.1%,25.0% and 3.1%,respectively. A novel subtype of aac(6')-Ⅰb was reported firstly. CONCLUSIONS There are high positive percentage of 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes in P. aeruginosa isolated from burned patients. P. aeruginosa resistance to aminoglycoside relates to the existence of 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes.

Objective To investigate the effects of contrast dose and region of interest (ROI) depth on quantitative analysis of liver by contrast-enhanced ultrasound (CEUS) during clinical application.Methods After bolus injection of contrast agent,the change of quantitative parameters [including echo mean(EM),rise time(RT),peak intensity(PI),mean transit time(MTT),area under the curve(AUC),time from peak to one half(TPH),wash in slope(WIS),time to peak(TTP)] of time-intensity curves were analyzed based on groups from different doses (1.0 ml and 1.6 ml) and different depth (＜30 mm,30-60 mm,and ≥60 mm).Results MTT and TPH were increased with dose increasing from 1.0 ml to 1.6 ml (P＜0.05).With the dose 1.0 ml,TPH,WIS,PI,AUC and MTT showed significant difference when the depth of the ROI changes (P ＜0.05),with the depth increased,TPH,WIS,PI,and AUC all decreases and MTT increases.For all the other parameters,no significant changes were found (P ＞0.05).Conclusions CEUS and its imaging process can directly influence the accuracy of the parameters from the quantitative analysis.Standardization of contrast agent with predefined dose and depth can potentially facilitate future clinical studies in liver CEUS.

Objective To investigate the application of contrast-enhanced ultrasonograhy CEUS in the early assessment of antiangiogenic treatment in metastatic colorectal cancer Methods Among 41 colorectal cancer patients with liver metastases enrolled in this study 25 patients received treatment of antiangiogenic treatment bevacizumab combined with chemotherapy BV group and 16 patients received chemotherapy only non-BV group To assess the treatment responses in the liver lesions between and within these two groups CEUS scans were performed before the treatment T0 or??baseline on the day 2 T1 and on the day 7 T2 after the treatment The percentage differences were evaluated according to the CEUS quantitative parameters such as rise time RT peak intensity PI mean transit time MTT area under the curve AUC time from peak to one half TPH wash in slope WIS and time to peak TTP Results On T1 both the RT and MTT increased within the BV group and decrease within the non-BV group when compared with their own baselines the differences were statistically significant P <0 05 On T2 the MTT increased within the BV group and decreased within the non-BV group when compared with their baselines the differences again were statistically significant P <0 05 All other parameters measured on T1 and T2 did not show a significant difference when compared with baselines Within the BV group the RT decreased and WIS and TPH increased for the responding subjects when compared with the non-responding subjects the differences were statistically significant P < 0 05 Within the non-BV group however no statistically significant differences were found for all parameters between the responding subjects and non-responding subjects P >0 05 Conclusions CEUS can be used to assess the treatment response in liver metastatic patients with colorectal cancer by monitoring the change of the tumor perfusion before and after the treatment.

Objective The effectiveness of Zhuang nationality medical lotus needle plus back cupping therapy ( Zhuang needle-cupping therapy) , Flixonase aqueous nasal spray and cetirizine tablets in treating allergic rhinitis (AR) was compared for the exploration of the therapeutic mechanism of Zhuang needle-cupping therapy. Methods A total of 200 recruited AR patients were randomly divided into four groups in the proportion of 1:1:1:1. The four groups were Zhuang needle-cupping therapy group, cetirizine group, Flixonase group and blank control group. The blank control group had no medication, and the patients of the other three medication groups were given the corresponding treatment. Ten days constituted one treatment course, and interval between two courses lasted one week. After two courses, the therapeutic effect was evaluated. The changes of specific IgE (S-IgE), leukotriene (LT), interleukin 4(IL-4), IL-9 mRNA, interferon gamma (IFN-γ), Thl / Th2 cells, and Th17 cytokine ( IL-17) were observed before and after treatment. Results ( 1) After two treatment courses, Zhuang needle-cupping therapy group had better therapeutic effect than cetirizine group , Flixonase group and blank control group, and the therapeutic effect of cetirizine group and Flixonase group was better than the blank control group (P<0.05). However, the therapeutic effect of cetirizine group was similar to that of Flixonase group ( P>0.05). ( 2) After treatment, the levels of S-IgE, LT, IL-9 mRNA, IL-4 and IL-17 were decreased, and IFN-γ and Th1/Th2 levels were increased in the three medication groups ( P<0.05 compared with those before treatment) . The differ ences of the laboratory indexes in the blank control group were insignificant before and after treatment ( P>0.05). The results of inter-group comparison after treatment showed that Zhuang needle-cupping therapy group had better effect on improving S-IgE, LT, IFN-γand Th1/Th2 than cetirizine group and Flixonase group (P<0.05). (3) During the trial, no adverse reaction was found. Conclusion Zhuang needle-cupping therapy exerts certain therapeutic effect for AR, and the mechanism may be related with the inhibition of S-IgE, LT, IL-9 mRNA and IL-17 expression, and with the regulation of Th1/Th2 imbalance by decreasing TH2 cytokine level and increasing Th1 cytokine level.

OBJECTIVE: To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency. METHODS: A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software. RESULTS: The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4). CONCLUSION The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.
Male ; Humans ; Aged ; Middle Aged ; Pedigree ; East Asian People ; Exons ; Introns ; Family ; Factor XII Deficiency/genetics* ; 3' Untranslated Regions ; Factor XII/genetics*

Objective:To investigate the risk factors and treatment outcome of recurrent Acanthamoeba keratitis (AK) after corneal transplantation. Methods:A serial case-observational study was carried out.Twenty-eight eyes of 28 patients with AK who underwent corneal transplantation in Shandong Eye Hospital from January 2012 to January 2019 were enrolled.All the eyes received corneal transplantation from failing to respond to topical and systemic anti- Acanthamoeba medical therapy, including 13 eyes that received penetrating keratoplasty (PKP) and 15 eyes that received lamellar keratoplasty (LKP). The corneal lesion was removed by a trephine with a diameter of 0.5 mm over infiltration area during PKP or LKP.The clinical features of recurrent AK were summarized, including recurrence time, site and signs, and the risk factors of AK recurrence were analyzed.Local and systemic anti- Acanthamoeba medical therapy was performed in all relapsed eyes, and secondary surgery was performed for the eyes with poor response to medication.The therapeutic outcome of recurrent AK was evaluated.The study adhered to the Declaration of Helsinki.This study protocol was approved by an Ethics Committee of Shandong Eye Hospital (No.201112). Results:In the 28 eyes, 7 eyes (25%) appeared recurrent AK after keratoplasty, including 2 eyes after PKP and 5 eyes after LKP.There was no significant difference in the recurrence rate between the two methods ( P=0.396). The recurrence rate of eyes that had used glucocorticoids drugs before operation was 57.14% (4/7), which was significantly higher in comparison with 14.29% (3/21) of eyes without glucocorticoids before surgery ( P=0.043). The recurrence rate of eyes with ulcer diameter ≥8.2 mm was 50.00% (5/10), which was significantly higher than 11.11% (2/18) of eyes with ulcer <8.2 mm ( P=0.036). The recurrent lesions began at the edge of implant bed accounted for 85.71% (6/7), and the recurrent lesions located below graft accounted for 14.29% (1/7). In 7 eyes with recurrent AK, 6 eyes were completely cured.Among recurrent AK eyes after LKP, 2 eyes were cured by long-term medical therapy, and 2 eyes were cured by extended-diameter LKP, and another 1 eye was cured by conjunctival flap covering surgery.One eye with recurrent AK after PKP was cured by extended-diameter PKP. Conclusions:The risk factors of recurrent AK after surgery are application of glucocorticoids before surgery and big lesions.Recurrent AK after surgery is curable by individualized therapy targeting to different clinical characteristics.