An effective RT-PCR method was developed to detect exogenous gene xylB transcript levels in Zymomonas mobilis CP4. Total RNAs without genomic DNA contamination were purified from wild-type and gene engineering strains, and were quantified to the same concentration. Then, cDNAs synthesis and PCR analysis of these samples were conducted by reverse transcription PCR. The optimal number of cycles was determined by observing amplification profile of target gene xylB and internal control gene 16s rRNA, and relative expression levels of xylB in various samples were analyzed by RT-PCR. The results indicated that the xylB transcript was not be detected in CP4, however that could be found in recombinant strains, in which xylB transcription abundance was similar. The enzyme assay furthermore confirmed that effective ex-pression of the target gene. The method provided a useful and rapid tool for detecting transcript levels of target genes from various samples of Z. mobilis.

Objective To investigate the relationship between the prevalence of isolated systolic hypertension (ISH) and the stages of chronic kidney disease (CKD) in chronic kidney disease outpatient clinic.Methods CKD patients of stages 1,2,3,4 and 5 were recruited (n=626).Based on office systolic pressure (SBP) and diastolic pressure (DBP),they were classified into four subtypes:normotension (＜ 140/90 mmHg),isolated diastolic hypertension (IDH,SBP ＜ 140 mmHg and DBP ≥ 90 mmHg),ISH (SBP≥ 140 mmHg and DBP ＜ 90 mmHg) and systolic-diastolic hypertension (SDH,SBP≥140 mmHg and DBP≥90 mmHg).Results The control rate of blood pressure was 86.4％,75.6％,65.3％,51.0％ and 37.0％ at CKD stage 1,2,3,4 and 5,respectively,which decreased with the advancement of CKD.There was a stepwise increase in the prevalence of ISH (0,9.2％,23.9％,28.6％ and 37.0％ at CKD stage 1,2,3,4 and 5,respectively) and SDH (4.5％,8.4％,8.0％,17.3％,21.9％ at CKD stage 1,2,3,4 and 5,respectively).Logistic regression analysis showed that age,diabetes and CKD stages were independent predictors of ISH.Compared with CKD stage 1-2,CKD stage 3,4 and 5 showed 2.388,2.697 and 5.980 folds risk in developing ISH.Conclusion The prevalence of ISH increases correspondingly with the advancement of stages of CKD,which may partially contribute to the increased cardiovascular mortality during the progress of CKD.

Objective To investigate whether there is any difference in aortic stiffness among different hypertension subtypes in patients with chronic kidney disease.Methods Six hundred and twenty-six patients with chronic kidney disease were included in the present analysis.They were classified into four groups:normotension (n =391) with systolic blood pressure (SBP) ＜ 140 mmHg and diastolic blood pressure (DBP) ＜ 90 mmHg; isolated systolic hypertension (ISH,n =141) with SBP≥ 140mmHg and DBP ＜ 90 mmHg; isolated diastolic hypertension (IDH,n =25) with SBP ＜ 140 mmHg and DBP≥ 90 mmHg; systolic-diastolic hypertension (SDH,n =69) with SBP≥ 140 mmHg and DBP≥ 90mmHg.Aortic stiffness was assessed by pulse pressure and pulse wave velocity.Results The IDH group had lower mean age than the other groups(P ＜ 0.01).The percentage of diabetes in the ISH group was higher than that in the other groups.The comparison of aortic stiffness showed that the ISH and SDH groups had higher aortic stiffness than the normotension and IDH groups (P ＜ 0.01),but no significant difference in aortic stiffness was observed neither between the normotension and IDH groups nor between ISH and SDH groups.Conclusion Aortic stiffness is significantly different among different hypertension subtypes,which may be an underlying cause for the different cardiovascular mortality among the hypertension subtypes.

PURPOSE: Primary low-grade thyroid-like papillary adenocarcinomas are extremely rare neoplasms that generally originate in the nasopharynx. Here, we describe a novel case of a 15-year-old Chinese girl who was diagnosed with low-grade thyroid-like papillary adenocarcinoma, including a brief review of the literature to reveal the clinicopathological features of low-grade thyroid-like nasopharyngeal papillary adenocarcinoma. MATERIALS AND METHODS: Immunohistochemistry was used to evaluate the expression of pan-cytokeratin (CKpan), cytokeratin (CK) 7, thyroid transcription factor 1 (TTF-1), vimentin, epithelial membrane antigen (EMA), thyroglobulin, CD15, S100, P40, CK20, CDX-2, glial fibrillary acidic protein (GFAP), and Ki-67. Additionally, in situ hybridization investigation was utilized to identify the presence of small Epstein-Barr virus (EBV)–encoded RNA. RESULTS: Histopathological analysis revealed florid proliferation of papillary structures lined by columnar epithelial cells with fibrovascular cores. Immunohistochemically, the neoplastic cells were positive for CKpan, CK7, TTF-1, vimentin, and EMA, but negative for thyroglobulin, CD15, S100, P40, CK20, CDX-2, and GFAP. The Ki-67–labeling index reached 5% in the most concentrated spot. In situ hybridization for EBV was negative. CONCLUSION: Due to the distinct rarity of low-grade thyroid-like papillary adenocarcinomaswith a favorable clinical outcome, a nationwide effort to raise public awareness of this neoplasm is required.
Adenocarcinoma, Papillary* ; Adolescent ; Asian Continental Ancestry Group ; Epithelial Cells ; Female ; Glial Fibrillary Acidic Protein ; Herpesvirus 4, Human ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Keratins ; Mucin-1 ; Nasopharynx ; RNA ; Thyroglobulin ; Thyroid Gland ; Transcription Factors ; Vimentin

Objective: To evaluate the effects of office blood pressure(OBP)combined with ambulatory blood pressure monitoring(ABPM)on the diagnosis of hypertension. Methods: The residents aged 35-79 years without hypertension history,whose casual OBP were 120~159 mm Hg/80~99 mm Hg,were enrolled from 4 communities of Hangzhou and Zhuji from 2015 to 2018. They were performed OBP measurements on other two days in 4 weeks and ABPM in a week. There were 2 criteria of OBP as elevated OBP on the first day or in 3 different days,and 4 criteria of ABPM as elevated mean BP in 24 hours, daytime, nighttime and either of the above time. Receiver operating characteristic(ROC)curve was employed to evaluate the effects of different OBP criteria combined with ABPM criteria on the diagnosis of masked hypertension(MH)and white-coat hypertension(WCH). Results: Taking 3-day-OBP as a golden standard,the 1-day-OBP with 4 ABPM criteria had the areas under the ROC curve(AUC)of 0.79-0.81,sensitivity of 57.58%-62.77% and specificity of 100.00% in MH;had the AUC of 0.95-0.98,sensitivity of 100.00% and specificity of 88.96%-96.80% in WCH. The Kappa values were all less than 0.6,known as low consistency. Taking either time of ABPM as a golden standard,24 hours,daytime and nighttime ABPM criteria with OBP had the AUC of 0.90-0.92,sensitivity of 79.17%-83.90% and specificity of 100.00% in MH(all Kappa>0.6),when with 1-day-OBP,the Kappa values were all more than 0.8,known as high consistency;had the AUC of 0.95-1.00,sensitivity of 100.00% and specificity of 89.54%-99.37% in WCH,the Kappa values of daytime ABPM were all more than 0.6,known as high consistency. Conclusions If limited by options, 1-day-OBP could be used instead of 3-day-OBP for detection of WCH or exclusion of MH yet with less accuracy; 24 hours or daytime ABPM instead of either time of ABPM was reliable.

OBJECTIVE: To investigate the efficacy of a novel artificial perfusate based on oxygen-carrying perfluoronaphthalene-albumin nanoparticles in normothermic machine perfusion (NMP) for preservation of porcine liver donation after cardiac death. METHODS: Artificial perfusate with perfluoronaphthalene-albumin nanoparticles was prepared at 5% albumin (w/v) and its oxygen carrying capacity was calculated. The livers of 16 Landrace pigs were isolated after 1 h of warm ischemia, and then they were divided into 4 groups and preserved continuously for 24 h with different preservation methods: cold preservation with UW solution (SCS group), NMP preservation by whole blood (blood NMP group), NMP preservation by artificial perfusate without nanoparticles (non-nanoparticles NMP group) and NMP preservation by artificial perfusate containing nanoparticles (nanoparticles NMP group). Hemodynamics, tissue metabolism, biochemical indices of perfusate and bile were monitored every 4 h after the beginning of NMP. Liver tissue samples were collected for histological examination (HE and TUNEL staining) before preservation, 12 h and 24 h after preservation. RESULTS: The oxygen carrying capacity of nanoparticles in 100 mL artificial perfusate was 6.94 μL/mmHg (1 mmHg=0.133 kPa). The hepatic artery and portal vein resistance of nanoparticles NMP group and blood NMP group remained stable during perfusion, and the vascular resistance of nanoparticles NMP group was lower than that of blood NMP group. The concentration of lactic acid in the perfusate decreased to the normal range within 8 h in both nanoparticles NMP group and blood NMP group. There were no significant differences in accumulated bile production, alanine aminotransferase and aspartate aminotransferase in perfusate between nanoparticles NMP group and blood NMP group (all P>0.05). After 24 h perfusion, the histological Suzuki score in blood NMP group and nanoparticles NMP group was lower than that in SCS group and non-nanoparticles NMP group (all P<0.05), and the quantities of TUNEL staining positive cells in blood NMP group and non-nanoparticles NMP group was higher than those in nanoparticles NMP group and SCS group 12 h and 24 h after preservation (all P<0.05). CONCLUSION Artificial perfusate based on oxygen-carrying nanoparticles can meet the oxygen supply requirements of porcine livers donation after cardiac death during NMP preservation, and it may has superiorities in improving tissue microcirculation and alleviating ischemia-reperfusion injury.
Swine ; Animals ; Liver Transplantation ; Organ Preservation ; Liver ; Perfusion ; Death ; Oxygen/metabolism*