Objective To construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18 of Taenia solium,to obtain purified recombinant protein TSO45W-4B-TSOL18 and to prepare rabbit antiserum against the recombinant fusion protein.Methods TSO45W-4B and TSOL18 encoding genes were connected with hydrophobic (Gly4Ser)3 linker.TSO45W-4B-TSOL18 fusion gene was synthesized and cloned into an expression vector pGEX-1λT to construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18.The positive recombinants were transformed into Escherichia(E.) coli ArcticExpress (DE3),and the expression of recombinant protein was induced with isopropylβ-D-thiogalactopyranoside (IPTG) and analyzed by sodium dodecyl sulfate polyacrylamide gelelectrophoresis (SDS-PAGE).The expressed recombinant fusion protein was purified through affinity chromatography.The purified recombinant protein was mixed with complete Freunds adjuvant at a dosage of 0.5 mg.Rabbit was intramuscularly and subcutaneously immunized in the hind leg and the back,respectively.After 2 weeks the rabbit was boosted with purified recombinant protein which was mixed with incomplete Freunds adjuvant at the same dosage.Rabbit can be boosted every 10 days until an adequate response was achieved.At the 6th day after the last immunization,blood was collected from the rabbit heart.Serum was separated to purify and prepare the antiserum.ELISA was applied to determine the titer of the antiserum and Western blot assay was used to determine the specificity of the antiserum.Results The size of the synthesized fusion gene TSO45W-4B-TSOL18 was 789 bp.The results of restriction enzyme digestion showed that the recombinant plasmid pGEX-TSO45W-4B-TSOL18 was successfully constructed.DNA sequencing showed that the size of the fusion gene TSO45W-4B-TSOL18 was 789 bp,which was consistent with expected result.As demonstrated by SDS-PAGE,relative molecular mass of the expressed recombinant fusion protein was approximately 55 × 103,and its purity was 85％ after purification with affinity chromatography.Titer of the antiserum against the recombinant protein was 1 ∶ 512 000 in ELISA assay and the specific rabbit antiserum against the purified recombinant protein TSO45W-4B-TSOL18 could specifically bind to the recombinant fusion protein in Western blot assay.The relative molecular mass of the specific band was about 55 × 103,which was consistent with expected size.Conclusions The recombinant plasmid pGEX-TSO45W-4BTSOL18 of Taenia solium is successfully constructed.High quality recombinant fusion protein TSO45W-4B-TSOL18 and high titer rabbit antiserum are successfully prepared.

Objective To explorethe effect of Bailingcapsule on epithelial-mesenchymal transition(EMT) inrats withadenine-in-duced tubulointerstitial fibrosis .Methods Tubulointerstitial fibrosis ani mal models were established and SDrats were dividedinto mo-del group (n=30) ,treatment group (n=30) andcontrol group(n=30) ,randomly .Experi mental rats were harvested at 7 w,12 w,17 wafter onset of experi ment and functional evaluations were performed. Histology ,i mmunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7) ,transforming growth factor-?1(TGF-?1)and a-smooth muscle actin (?-SMA) in kidneys at three ti me points mentioned above ,respectively .Results Compared with controlgroup ,24 h urinary proteinin model grouplost increasingly and significantly difference appeared at three ti me points relative to controlgroup(P0 .05) rel-ative to control group.There was significant difference at 12 wand 17 w(P

Objective To explore the effect of Bailing capsule on epithelial-mesenchymal transition( EMT) in rats with adenine-in-duced tubulointerstitial fibrosis. Methods Tubulointerstitial fibrosis animal models were established and SD rats were divided into mo-del group ( n = 30), treatment group ( n = 30) and control group( n = 30), randomly. Experimental rats were harvested at 7 w, 12 w,17 w after onset of experiment and functional evaluations were performed. Histology, immunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7), transforming growth factor-β1 (TGF-β1 )and a-smooth muscle actin (α-SMA) in kidneys at three time points mentioned above, respectively. Results Compared with controlgroup, 24 h urinary protein in model group lost increasingly and significantly difference appeared at three time points relative to controlgroup ( P ＜ 0.01 ). Urinary NAG in model group was markedly higher than that in control group from 7 w after onset (P ＜ 0.01 ) andwas increasingly raised at 12 w and 17 w (P＜0.01). The value of blood BUN and Cr in model group increased at 7 w (P＞0.05) rel-ative to control group. There was significant difference at 12 w and 17.w (P ＜ 0.01 ). Histologically, kidneys in model group, at 7 w,exhibited tubular casts and gently tubular dilation, granuloma in cortex, mononuclear cells infiltration in tubulointerstitial areas, andmild interstitial fibrosis. At 12 w, the degree of tubular injury and tubulointerstitial fibrosis gradually aggravated. Up to 17 w, diffusetubular dilation or atrophy was observed and focal tubules disappear. Diffuse interstitial fibrosis was exhibited. In normal kidneys, im-munohistochemistry suggested that the light expression of BMP-7 was detected in proximal renal tubular epithelial cells and marked ex-pression was identified in distal tubule, collecting duct, and renal tubular epithelial in junction area between cortex and medulla. How-ever, the expression of BMP-7 in kidneys of model group significantly decreased with increasing tubulointerstitial fibrosis and was nega-tive correlation with the expression of TGF-β1(r = -0. 981 P＜0.01) and α-SMA (r= -0.975 P＜0.01). Bailing capsule ad-ministration protected the expression of BMP-7 and reduced TGF-β1 and α-SMA expression before 12 w(P＜ 0.01 ). Conclusions Ourstudy shows an anti-fibrotic reno-protective function of Bailing capsule in rats with tubulointerstitial fibrosis via prevention of epithelial-mesenchymal transition at early stage. However, the beneficial effect lost with increasing tubulointerstitial fibrosis.

Objective To investigate the morphological features of normal lumbar dorsal root ganglia using a three-dimensional(3D)coronal MR imaging.Methods One hundred and fifteen volunteers were included.Ages ranged from 15 to 75 years,with a mean of 40 years.Coronal 3D fast field echo(FFE) with water selective excitation(Proset)MR examination of 1150 dorsal root gangha were underwent at nerve root levels from L1 to L5.The source coronal images were further reconstructed into a series of rotational alignment coronal images with an interval angel of 12 degree using maximum intensity projection(MIP) technique.All DRGs of bilateral spinal nerve from L1 to L5 were morphologically analyzed on the original and MIP images including qualitative evaluation of the location,signal intensity,architecture and quantitative dimensional measurement.Results There were 225,225,219,210 and 160 foraminal ganglia from L1 to L5 level,respectively.The incidence of intraspinal ganglia from L3 to L5 gradually increased with a maximum at L5 level of 29.1%(X~2=188.371,P

A simple,rapid and sensitive liquid chromatography-mass spectrometry (LC-MS) method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic etherisopropanol (2:1) and the extraction was performed on a Kromasil C18 column (150 mm×4.6mm,5 μm) with the mobile phase of methanol-water (41:59,v/v) within a run time of 6.0min.The analyte was monitored with positive electrospray ionization (ESI) by selected ion monitoring (SIM) mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard (IS) geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.

A simple,rapid and sensitive liquid chromatography-mass spectrometry（LC-MS）method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic ether-isopropanol（2∶1）and the extraction was performed on a Kromasil C18 column（150 mm×4.6 mm,5 μm）with the mobile phase of methanol-water（41∶59,v/v）within a run time of 6.0 min.The analyte was monitored with positive electrospray ionization（ESI）by selected ion monitoring（SIM）mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard（IS）geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.

Objective To investigate MR imaging features of femoral marrow in treated ?-thalassemia major.Methods MR imaging of the proximal femoral marrow was performed in 35 cases of ?-thalassemia major and 45 age-and sex-matched normal children as control.Coronal images of femoral marrow with the techniques of spin echo and fast field echo(FFE)were obtained.On T_1-weighted imaging the red and yellow femoral marrow were judged and marrow distribution was classified into five groups.The hemosiderosis of marrow was judged on the basis of signal intensity of marrow on FFE imaging.The marrow distribution classification and the hemosiderosis on MR imaging were correlated with clinical features.Results On FFE,marrow hemosiderosis occurred in 15 patients with a marked hypo-intensity signal and was related to the age(P=0.032).On T_1-weighted imaging,the femoral marrow in 35 patients was classified as groupⅢand IV,while the marrow distribution was groupⅠorⅡin all normal children,there was statistically significant difference(P

Objective To explore the distribution rule of metastatic lymph node in nasopharyngeal carcinoma (NPC) by magnetic resonance imaging (MRI).Methods 315 histopathologically proved NPC patients were studied retrospectively.All patients had had their nasopharynx scanned by MRI with plain and contrast enhanced sequences.The distribution of lymph node was divided into six cervical levels plus retro- pharyngeal nodes(RN) according to RTOG guidelines proposed in 2003.Results 254 out of 315 patients (80.6%) had lymph node involvement,with 81 in the right neck alone,72 left neck alone,and 101 both necks;73 in RN alone,21 neck node alone,and 160 both necks and RN node.Skip metastasis was found in only 4 patients (1.6%).There was significant difference in BN metastasis between the primary tumor be- ing located merely on the superior/posterior wall and lateral wall (78% vs 49%,P＜0.01).The incidence of lymph node metastasis in T1,T2,T3 and T4 patients was 73.5%,91.2%,71.9%,73.5% (P＞0.05), respectively,without significant difference between early or advanced T stage in node distribution (P＞0.05).Conclusions The incidence of lymph node metastasis is high in nasopharyngeal carcinoma,with retropharyngeal node being the most commonly involved,but the incidence of skip metastasis is very low. There is no significant difference between T stage and the incidence of lymph node metastasis.So is the dis- tribution of metastatic node.

BACKGROUND: Measurement uncertainty characterizes the dispersion of the quantity values attributed to a measurand. Although this concept was introduced to medical laboratories some years ago, not all medical researchers are familiar with it. Therefore, the evaluation and expression of measurement uncertainty must be highlighted using a practical example. METHODS: In accordance with the procedure for evaluating and expressing uncertainty, provided by the Joint Committee for Guides in Metrology (JCGM), we used plasma glucose (Glu) as an example and defined it as the measurand. We then analyzed the main sources of uncertainty, evaluated each component of uncertainty, and calculated the combined uncertainty and expanded uncertainty with 2 budgets for single measurements and continuous monitoring, respectively. RESULTS: During the measurement of Glu, the main sources of uncertainty included imprecision, within-subject biological variance (BVw), calibrator uncertainty, and systematic bias. We evaluated the uncertainty of each component to be 1.26%, 1.91%, 5.70%, 0.42%, and -2.87% for within-run imprecision, between-day imprecision, BVw, calibrator uncertainty, and systematic bias, respectively. For a single specimen, the expanded uncertainty was 7.38% or 6.1+/-0.45 mmol/L (kappa=2); in continuous monitoring of Glu, the expanded uncertainty was 13.58% or 6.1+/-0.83 mmol/L (kappa=2). CONCLUSIONS: We have demonstrated the overall procedure for evaluating and reporting uncertainty with 2 different budgets. The uncertainty is not only related to the medical laboratory in which the measurement is undertaken, but is also associated with the calibrator uncertainty and the biological variation of the subject. Therefore, it is helpful in explaining the accuracy of test results.
Blood Chemical Analysis/methods/standards ; Clinical Chemistry Tests/*methods/standards ; Glucose/*analysis/standards ; Humans ; Models, Statistical ; Quality Control ; *Uncertainty

BACKGROUND: Measurement uncertainty characterizes the dispersion of the quantity values attributed to a measurand. Although this concept was introduced to medical laboratories some years ago, not all medical researchers are familiar with it. Therefore, the evaluation and expression of measurement uncertainty must be highlighted using a practical example. METHODS: In accordance with the procedure for evaluating and expressing uncertainty, provided by the Joint Committee for Guides in Metrology (JCGM), we used plasma glucose (Glu) as an example and defined it as the measurand. We then analyzed the main sources of uncertainty, evaluated each component of uncertainty, and calculated the combined uncertainty and expanded uncertainty with 2 budgets for single measurements and continuous monitoring, respectively. RESULTS: During the measurement of Glu, the main sources of uncertainty included imprecision, within-subject biological variance (BVw), calibrator uncertainty, and systematic bias. We evaluated the uncertainty of each component to be 1.26%, 1.91%, 5.70%, 0.42%, and -2.87% for within-run imprecision, between-day imprecision, BVw, calibrator uncertainty, and systematic bias, respectively. For a single specimen, the expanded uncertainty was 7.38% or 6.1+/-0.45 mmol/L (kappa=2); in continuous monitoring of Glu, the expanded uncertainty was 13.58% or 6.1+/-0.83 mmol/L (kappa=2). CONCLUSIONS: We have demonstrated the overall procedure for evaluating and reporting uncertainty with 2 different budgets. The uncertainty is not only related to the medical laboratory in which the measurement is undertaken, but is also associated with the calibrator uncertainty and the biological variation of the subject. Therefore, it is helpful in explaining the accuracy of test results.
Blood Chemical Analysis/methods/standards ; Clinical Chemistry Tests/*methods/standards ; Glucose/*analysis/standards ; Humans ; Models, Statistical ; Quality Control ; *Uncertainty