Aims To construct the N-terminal Strep-tagged ( NS-tagged) fusion protein expression vector, and to apply the vector to express NS-tagged fusion proteins of Chlamydia RNA polymerase subunit. Meth-ods By using PCR method, NS fusion protein tag and a new multiple cloning sites (MCS) were inserted into pET21c-DH plasmid by primers to replace the original T7 protein tag and MCS. The newly introduced Not I cutting site was chosen for self-ligation of PCR prod-uct. Then, the cyclized PCR product was transformed into DH-5α competent cells. The positive clones were selected by PCR and sequencing. To get NS-tagged fu-sion proteins of chlamydial RNA polymerase subunits, the α, β and β′ subunits were inserted between BamH I and Sal I cutting sites of the newly constructed ex-pression vector. Then, the NS-α, NS-β and NS-β′ ex-pression vectors were transformed into Arctic Express expression cells. The fusion protein expression statuses of transformed cells were identified by Commassie blue staining and Western blot. Results The NS-tagged fusion protein expression vector pET21c-NS-MCS was successfully constructed, and NS-α, NS-β and NS-β′fusion proteins were obtained by using this newly con-structed expression vector. Conclusions In this pro-ject, we constructed an NS-tagged fusion protein ex-pression vector and applied it to express NS-α, NS-βand NS-β′ fusion proteins. Our study can lay a solid foundation for the study of transcriptional regulation of Chlamydia genes.

Humans ; Infant, Newborn ; Male ; Protein C ; metabolism ; Protein C Deficiency

OBJECTIVE:To investigate the effects of sevoflurane combined with remifentanil on intraoperative related indica-tors and quality of postoperative recovery in patients with general anesthesia. METHODS:52 patients with elective abdominal sur-gery were randomly divided into observation group and control group. All patients were treated with etomidate 0.3 mg/kg+atracuri-um 0.5 mg/kg+remifentanil 1μg/kg for anesthesia induction;then observation group was received sevoflurane by inhalation and con-trol group was propofol by infusion. The SBP,DBP,HR and BIS in 2 groups were recorded before anesthesia induction(T0),after anesthesia induction(T1),time of intubation(T2),skin incision(T3),10 minutes after establishing pneumoperitoneum(T4),end of pneumoperitoneum(T5) and extubation(T6),respectively. Operation and anesthesia duration,recovery time and extubation time, MMSE score and incidence of adverse reactions were recorded. RESULTS:SBP and HR in T6 and DBP in T4-6 in 2 groups were sig-nificantly higher than T0 of same group,BIS in T1-5 was significantly lower than T0,the differences were statistically significant(P<0.05),however,there was no significant difference between 2 groups (P>0.05). Recovery time and extubation time in observa-tion group were significantly lower than control group(P<0.05),there was no significant difference in the operation and anesthesia duration between 2 groups(P>0.05). MMSE scores in 2 groups were significantly lower than before after 0.5 h and 1 h of extuba-tion,and control group was lower than observation group(P<0.05 or P<0.01);MMSE scores in 2 groups were significantly high-er than 0.5 h and 1 h after extubation(P<0.05),however,there was no significant difference between 2 groups and before(P>0.05). There was no significant difference in the incidence of adverse reactions between 2 groups(P>0.05). CONCLUSIONS:Sevoflurane combined with remifentanil has satisfactory anesthesia,sevoflurane has better controllability,with good safety.

Objective To investigate the mechanism of fluoroquinolone resistance in clinical isolates of Enterococcus faecium. Methods The MICs of six fluoroquinolones(norfloxacin,ciprofloxacin,ofloxacin,levofloxacin,gatifloxacin and moxifloxacin) against 35 clinical isolates of E.faecium from eight hospitals in Tianjin were determined by agar dilution method in the absence or presence of multidrug resistance efflux pump inhibitor reserpine.The quinolone-resistance determining region(QRDR)of parC and gyrA were amplified and sequenced.Results No less than twofold decrease in MIC values of the six fluoroquinolones in the presence of reserpine was observed in 35,29,1,0,6 and 2 of the 35 strains of E.faecium respectively.One fluoro- quinolone-susceptible isolate and five fluoroquinolone-resistant isolates were selected randomly to analyze the QRDR of parC and gyrA.All five fluoroquinolone-resistant isolates had single amino acid alteration in both GyrA and ParC.Ser-80 in ParC was substituted by lie(4 isolates)or Arg(1 isolates).Glu-87 in GyrA was replaced by Lys(2 isolates)or Gly(2 isolates). The other one had an Ser-83-to-Ile substitution.The one fluoroquinolone-suseeptible isolate had no alteration in the QRDR of either ParC or GyrA.Conclusions Both target alteration and active efflux are responsible for the resistance to fluoroquinolone in clinical isolates of E.faecium.

Objective To investigate the feasibility of ~1H-MR spectroscopy(~1H-MRS)imaging to quantitatively detect fatty liver.Methods Twenty patients with fatty liver and 11 healthy volunteers underwent plain CT scan,conventional MR imaging and ~1H-MRS analysis.The blood lipid and liver function were tested on the same day as the MR examination.~1H-MRS sequence measured the peaks of H_2O and lipid,and the areas under the peaks.The relative contents of the lipid compound were calculated,and compared with the results of CT scan and liver function tests.Results The CT values of the normal group and the fatty liver group were(59?9)HU and(24?11)HU respectively.On ~1H-MRS a protruding high H_2O peak and a flat low lipid peak were observed in the normal group,while the protruding high H_2O peak and a high lipid peak appeared in the fatty liver group.The values of lipid peak in the normal group and the fatty liver group were(0.05?0.01)?10~5,(0.70?0.24)?10~5 respectively(t=4.32,P0.05),the areas under the lipid peak were(1.36?0.73)?10~9、(2.35?1.15)?10~9 respectively(t=5.21,P0.05).Conclusion ~1 H-MRS imaging is feasible to quantitatively detect liver fat and is a non-invasive method for detecting early fatty liver.

Objective To explore the value of 18F-FDG PET/CT on the assessment of chemotherapy response in patients with diffuse large B-cell lymphoma (DLBCL). Methods 18F-FDG PET/CT was performed before and after 4 cycles of chemotherapy( R-CHOP or CHOP protocol) in 53 patients with DLBCL. The patients were divided into 3 groups: complete response group, partial response group and no response group. The therapeutic response was assessed by comparing post-treatment 18F-FDG PET/CT with pre-treatment PET/CT. Complete remission (CR) rate at the end of chemotherapy was calculated. χ2 test was performed with software SPSS 13.0. Results CR rates of complete response group, partially response group and no response group were 88.5% (23/26), 73.3% (11/15) and 8.3% (1/12), respectively (χ2=23.548, P=0.000). CR rates of the complete and partially response groups were significantly higher than those of no response group (χ2=22.656, P=0.000; χ2=11.407, P=0.001). Conclusion 18F-FDG PET/CT may be useful for the assessment of chemotherapy response in DLBCL.

Objective To explore the mechanisms by which hydrogen sulfide (H2S) regulates collagen metabolism during hypoxic pulmonary vascular structural remodeling. Methods Wistar rats were randomly divided into control group (n=6), hypoxia group (n = 6) and hypoxia+NaHS group(n= 6) H2S content in plasma was measured with spectrophotometry. The expression of collagen I was detected by immunohistochemistry. The expression of tissue inhibitor of metalloproteinase-1(TIMP-1) mRNA was detected with situ hybridization Results Compared with hypoxia group, H2S content in plasma increased significantly and the expressions of collagen I and TIMP-1 mRNA in pulmonary arteries were down- regulated in hypoxia + NaHS group. The H2S content in plasma negatively correlated with TIMP-I mRNA expression. Conclusions H2S increases the degradation of collagen in small and media pulmonary arteries of rats under hypoxia by decreasing the synthesis of TIMP-1, and therefore, attenuats the collagen remodeling in pulmonary arteries of hypoxic rats.

Objective To investigate the value of diffusion kurtosis imaging (DKI) in differential diagnosis of parotid gland disease and diagnosis of parotid adenolymphoma (PAL).Methods DKI and DWI data of 57 patients with parotid gland disease were etrospectively analyzed.Totally 57 cases were divided into infectious lesions group (n=10),pleomorphic adenoma group (n=19),PAL group (n=14),other benign parotid tumor group (n=4) and malignant parotid tumor group (n=10).Contralateral normal parotid glands in 19 patients with unilateral parotid gland lesions were treated as control group.The quantitative parameters including kurtosis concerning parameters (K Krad,Kax),diffusivity concerning parameters (D Drad,Dax),fractional anisotropy (FA) and conventional apparent diffusion coefficient (ADC) values were retrospectively reviewed.The binary Logistic regression method was used to confirm parameters with significant difference in diagnosing PAL.And Logistic regression equation was constructed to diagnose PAL.ROC analysis was conducted to evaluate the diagnostic value of the confirmed parameters and the Logistic regression equation.Results Significant difference of the parameters including K Krad,Kax,D Drad,Dax,FA and ADC values were found among different groups (all P＜0.05).ROC analysis demonstrated a higher area under the curve (AUC) for FA+Kax [0.88±0.06 (0.79-0.94)] than Kax[0.80±0.07 (0.70-0.88)] and FA [0.63±0.10 (0.52-0.73)],respectively (both P＜0.05).The sensitivity,specificity,accuracy,positive predictive value and negative predictive value was 71.43％,95.78％,91.77％,76.92％ and 94.44％.Conclusion DKI showed high diagnostic capacity in differential diagnosis of parotid gland disease.The combination of FA and Kaxcan improve the diagnostic accuracy in diagnosis of PAL.

Animals ; Animals, Newborn ; Brain ; blood supply ; metabolism ; pathology ; Carrier Proteins ; genetics ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Gene Expression ; Hypoxia-Ischemia, Brain ; genetics ; pathology ; In Situ Hybridization ; Protein Tyrosine Phosphatase, Non-Receptor Type 13 ; Protein Tyrosine Phosphatases ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

Objective:To explore the pathogenesis of coronary artery lesions in Kawasaki disease(KD) by detecting expression of CD40L on T-cells from patients with KD.Methods:Blood samples were collected from 26 patients with KD before and after intravenous immunoglobulin(IVIG) treatment. Age-matched febrile 16 children with various diseases were studied in parallel as controls. Age-matched normal control 15 children were studied as controls. CD40L expression on T-cells was detected by flow cytometry, soluble E-selection and soluble CD40L levels were measured by enzyme-linked immunosorbent assay.Results:CD40L expression on CD4+T-cells and soluble E-selection levels were significantly higher in patients with KD than that in the febrile control(FC) group and normal group(P