Objective: To investigate the relationship between prognosis of patients with primary clear cell renal cell carcinoma (ccRCC) after radical nephrectomy and expression of tumor metastasis-associated gene CD99 and to analyze the prognostic factors of ccRCC paiients. Methods: Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of CD99 in primary ccRCC tissues and their corresponding adjacent tissues. The prognosis and risk factors of survival time of patients were studied by follow-up investigation, and the main risk factors were screened by Cox hazard regression model. Results: Compared with the adjacent renal tissues, 73.5% ccRCC tissues had up-regulated CD99 expression, with significant difference found between the two groups (P=0.000). Cox hazard regression model showed that high CD99 expression in ccRCC tissues was not a survival risk factor of ccRCC patients after radical nephrectomy (HR=0.14, 95%CI[0.01, 2.15]); and age (HR=1.18, 95%CI[1.01, 1.38]), TNM stages (HR=51.91, 95%CI[4.31, 625.87]), diabetes (HR=59.94, 95%CI[2.21, 1 627]) and hypertension (HR=47.72, 95%CI[1.37, 1 670]) were the major risk factors for the survival of patients after radical nephrectomy. The 1-year and 2-year survival rates of ccRCC patients in TNM stage I were significantly higher than those in TNM stage II-IV, respectively (100.0% vs 60.0%, P=0.004; 93.8% vs 8.3%,P=0.000). Conclusion: The expression of tumor metastasis-related gene CD99 may not be associated with the prognoses of ccRCC patients. Age, TNM stage, diabetes and hypertension are the major risk factors of prognosis after resection of ccRCC.

Objective: To predict the B cell epitopes of tumor-associated protein EIF4G1 subtypes. Methods: The sequences of all the protein subtypes of EIF4G1 were retrieved from NCBI protein database. Based on single parameter evaluation, including hydrophilicity, flexility, antigenicity, the B cell epitopes of the EIF4G1 protein subtypes were predicted using NPS@ structure software and ABCpred software. Results: EIF4G1 protein had five subtypes. The variation of the five different EIF4G1 subtypes was limited within a 300aa region. We identified eight epitopes locating in or near 14-19, 21-27, 52-61, 106-112, 113-139, 183-189, 201-216, and 217-224, which can be used to identify specific B cell epitopes of different protein subtypes. Conclusion: B cell epitopes of EIF4G1 protein subtypes do exist, and they may be used for the protein subtypes evaluation and early diagnosis of tumor patients using artificially-produced matched peptides.

Objective To investigate the effects of the expressions of thymidine kinase 1 (TK1) and Ki67 alone or their combination on the recurrence and metastasis of breast cancer.Methods Sixty-five samples were selected from the Second Affiliated Hospital of Guangzhou Medical University from March 2005 to June 2007,which were resected by surgical operation and confirmed as breast carcinoma by pathology.They were individed into two groups including 37 cases with recurrence or metastasis in 5 years (group A),28 cases without recurrence or metastasis in 5 years (group B).The expressions of TK1 and Ki67 in the two groups were detected by immunohistochemical staining assay.Then,Kaplan-Meier assay was used to describe survival curve.Results The positive expression rate of TK1 in group A was 91.8％,which was dramatically higher than that in group B 67.8％ (x2 =6.116,P =0.013).The positive expression rates of Ki67 in group A and B were 78.4％ and 42.9％ (x2 =8.635,P =0.003).The positive expression rates of TK1 combined with Ki67 in group A and B were 67.6％ and 39.3％ (x2 =5.159,P =0.023).Moreover,disease free survival of patients with positive expression of TK1 combined with Ki67 decreased significantly,compared with patients with positive expression of TK1 or Ki67 alone (x2 =6.137,P =0.046).Conclusion Positive expression of TK1 combined with Ki67 is the high risk factor of the reccurence or metastasis of breast carcinoma,and indicates poorer prognosis compared with positive expression alone.

Objective To establish an assay for the detection of Streptococcus pneumoniae by real-time fluorescence quantititive polymerase chain reaction (PCR).Methods Special primers and probe for the autolysin A (lytA)gene were designed.The sensitivity and specificity of primers and probe were studied,and cut-off of cycle threshold was assayed.158 clinical specimens were confirmed by real-time fluorescence quantitative PCR and bacterial culture method.Results Primer and probe design for LytA gene could sensitively detect serotype Streptococcus pneumoniae strains of common pathogenic,and the sensitivity was 100 copies.Among 35 strains of Streptococcus pneumoniae,34 cases were detected to be positive for Streptococcus pneumoniae by real-time fluorescence quantitative PCR,while 1 case was detected to be negative;among 15 strains of non-Streptococcus pneumoniae, all were detected to be negative.Among the 158 clinical sputum specimens,34 cases with Streptococcus pneumoniae were detected by real-time fluorescence quantitative PCR,while only 10 cases with Streptococcus pneumoniae were detected by the culture method.White blood cells count and time in hospital of cases with Streptococcus pneumoniae were higher than those of cases without Streptococcus pneumoniae (P <0.05 ). Conclusion Real-time fluorescence quantitative PCR is a sensitive and specific assay for the detection of Streptococcus pneumoniae.It can be used for the diagnosis of Streptococcus pneumoniae.

Objective To evaluate the role of chemokine CXCL12 in the spinal cord in the development of bone cancer pain (BCP) in rats and the relationship with microglial activation.Methods Thirty-two female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups (n =8 each) using a random number table:sham operation group (group S),BCP group (group B),BCP + CXCL12 neutralizing antibody group (group BC),and BCP + IgG control antibody group (group BI).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension (4 × 105 cells/ml) 5 μl into the bone marrow of the right tibia of rats anesthetized with chloral hydrate in B,BC and BI groups,while the equal volume of normal saline was injected instead in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,CXCL12 neutralizing antibody 10 μg/15 μl was intrathecally injected once a day in group BC,while IgG control antibody 10 μg/15 μl was intrathecally injected once a day in group BI.Before injection of mammary gland cancer cells (T0) and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells (T16),paw withdrawal threshold to mechanical stimulation (PWMT) was measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for determination of Iba-1 (pan-microglial marker) expression in spinal dorsal horn using immunofluorescence after PWMT measurement at T6.Results Compared with S group,PMWT was significantly decreased at T2-6,and Iba-1 expression was up-regulated at T6 in B,BC and BI groups (P ＜ 0.01).Compared with B group,PMWT was significantly increased at T5,6 and Iba-1 expression was down-regulated at T6 in BC group (P ＜ 0.01).Conclusion Chemokine CXCL12 in the spinal cord is involved in the development of BCP,and microglial activation is involved in the mechanism.

Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.

The objective of this study is to propose a more accurate and faster MTT [3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide] colorimetric assay (MCA) for quantitative measurement of polypeptide bacteriocins in solutions with nisin as an example. After an initial incubation of nisin and indicator bacterium Micrococcus luteus NCIB 8166 in tubes, MTT was added for another incubation period. After that, nisin was quantified by estimating the number of viable bacteria based on measuring the amount of purple formazan produced by cleavage of yellow tetrazolium salt MTT. Then MCA was compared to a standard agar diffusion assay (ADA). The results suggested a high correlation coefficient (r(2)=0.975+/-0.004) between optical density (OD) and the inhibitory effect of nisin on a bacterial strain Micrococcus luteus NCIB 8166 at a range of 0.125-32 IU/ml. The MCA described in this study was very quick. Quantification of nisin took only 7-8 h and the detection limit was at the level of 0.125 IU/ml when compared to 12 IU/ml and 24-28 h for ADA. The MCA provides an accurate and rapid method for quantification of nisin in solutions and is expected to be used for quantification of other antimicrobial substances.
Bacteriocins ; analysis ; metabolism ; Colorimetry ; methods ; Immunodiffusion ; Micrococcus luteus ; metabolism ; Nisin ; Regression Analysis ; Tetrazolium Salts ; analysis ; Thiazoles ; analysis ; Time Factors

Objective To determine the pharmacokinetic interaction between cefalor and bromhexine in healthy Chinese volunteers. Methods Twelve subjects received a cefaclor (CEF) treatment, a bromhexine (BHX) treatment, and a co-treatment of CEF and BHX with a 3 × 3 Latin square design. The wash-out time between periods was 14 days. The plasma and urine drug concentrations of CEF and BHX were detected by HPLC-UV and LC/MS, respectively. Results All the 12 volunteers completed the study. There were no significant differences in AUC0-t and Cmax of CEF in logarithm between the single administration group of CEF and the co-administration group of CEF with BHX. Two one sided t-test showed that CEF was bioequivalent in the 2 groups. There were no significant differences in tmax, MRT, t1/2, and Clr between the 2 groups. Vd/F was significantly lower in the single CEF group than in the co-administration group of CEF and BHX. There were no significant differences of AUC0-t and Cmax of BHX in logarithm between the single administration group of BHX and the co-administration group of BHX with CEF. Two one sided t-test showed that BHX was bioequivalent in the 2 groups. There were no significant differences in tmax, MRT, t1/2, Vd/F, and Clr between the 2 groups. Conclusion There is no significant pharmacokinetic parameter change in the drug absorption, metabolism, and excretion, but Va/F of CEF significant increases in the co-administration of CEF with BHX. The co-administration of CEF and BHX has no adverse drug interaction. The increase of Vd/F may be a favorable drug interaction, which may be the mechanism of the synergistic effect of the 2 drugs.

Background and purpose:The application of intensity-modulated radiotherapy (IMRT) has improved the local control rate of nasopharyngeal carcinoma greatly, which changed the predictive value of T classiifca-tions of TNM staging system. This study aimed to validate the predictive effect of T classiifcations in the 7th Union for International Cancer Control (UICC) staging system and discuss the simpliifcation of T classiifcations.Methods:We retrospectively reviewed the clinical data of 641 primary nasopharyngeal carcinoma patients at our center from January 2007 to June 2011. We evaluated the predictive effect of T classiifcations by Kaplan-Meier method and Cox regression model.Results:The 5-year overall survival (OS), local relapse-free survival (LRFS), progression-free survival (PFS) and distant metastasis free survival (DMFS) were 85.4%, 88.5%, 78% and 87.1%, respectively. The 5-year OS of T1, T2, T3 and T4 categories were 91.6%, 85.3%, 90.1% and 76.5%, respectively; LRFS were 93%, 85.3%, 91.5% and 84.4%; PFS were 88.2%, 77.3%, 80.8% and 70.9%; DMFS were 95.1%, 88.9%, 88.2% and 81.3%, respectively. The difference in survival curves between T1, T2 and T3 were not signiifcant (P>0.05). However, several prognostic indexes were signiifcantly different between T4 and T1, T2, T3. We merged the T1, T2 and T3 classiifcations as new T1, and the T4 classiifcation as new T2. The 5-year OS of new T1 and T2 were 89.1% and 76.5% (P=0.001); LRFS were 90.1% and 84.4% (P=0.028); PFS were 81% and 70.9% (P=0.001); DMFS were 90.8% and 81.2% (P=0.002). The survival curves were substantially separated. The simpliifed T classiifcations had obvious advantages when separately analyzed in different N stages.Conclusion:In the era of IMRT, the predictive effect of T classiifcations of the 7th UICC staging system has diminished. The simpliifcation of T classiifcations can ift with the new treatment and provide a better surviv-al prediction.

Objective To investigate the risk factors and prognosis of the patients with malignant solid tumors complicated with the disseminated intravascular coagulation (DIC). Methods The clinical data of 54 malignant solid tumors patients complicated with DIC in Shanxi Provincial People's Hospital from January 2004 to December 2014 were analyzed retrospectively, which were compared with the malignant tumor patients without DIC. Multivariate logistic regression analysis was used to explore the risk factors of solid tumor complicated with DIC, and the effect of concurrent DIC on the prognosis of patients was analyzed. Results Multiple factor analysis showed that advanced tumor (OR = 0.252, P = 0.019), concurrent hypoproteinemia(OR=0.119,P=0.005),concurrent infection(OR=0.122,P=0.003)were the independent risk factors of the malignant solid tumor patients complicated with DIC. The case fatality rate of the patients with DIC was 85.2 % (46/54), which was higher than that of the control group (7.4 %, 4/54), and the difference was statistically significant(χ2=65.69,P <0.001).Conclusion Early detection of malignant solid tumors, positive correction of hypoproteinemia, and the effective control of infection as soon as possible can help to prevent the occurrence of DIC and reduce the death caused by DIC.