Objective To investigate the age-related change of secretory leukocyte protease inhibitor (SLPI) content in stimulated whole saliva. Methods Ninety-four healthy adults aged 20-94 yrs were enrolled. Samples of whole stimulated saliva were collected from all subjects, the SLPI content was analyzed by enzyme-linked immunosorbent assay (ELISA). Results The flow rate of the SLPI in older groups was lower than in the young group (P0. 05) , but the contents of SLPI secretion per time-unit of the three older groups[(2. 12?1. 70), (2. 17?2. 65) , (1. 91?2. 51) ng/min, all P

Objective To evaluate the effect of gender matching on the outcomes of livingdonor renal transplantation.Methods A total of 419 cases of living-donor renal transplantation in our center were divided into male-donor-male-recipient (MDMR) group,male-donor-female-recipient (MDFR) group,female-donor-male-recipient (FDMR) group,female-donor-female-recipient (FDFR)group.The outcomes including graft and patient survival,acute rejection and renal function were analyzed retrospectively.Results Compared to MDMR group,MDFR group and FDFR group had lower Scr [(80.7±17.9),(87.4±21.9) μmol/L vs (120.3±72.5) μmol/L,all P ＜ 0.05] and uric acid (UA) [(318.1 ± 86.4),(303.5 ± 66.9) μmol/L vs (358.4 ± 77.8) μmol/L,P ＜ 0.05] 6 months after operation.Compared to MDFR group,FDMR group had higher Scr[(117.7±27.4) μmol/L vs (80.7±17.9) μmol/L,P ＜ 0.01],UA [(371.0±92.4) μmol/L vs (318.1±86.4) μmol/L,P ＜ 0.05] and lower glomerular filtration rate (GFR) [(70.4± 17.8) ml/min vs (79.6± 18.9) ml/min,P ＜ 0.05].Compared to FDMR group,FDFR group had lower Scr [(87.4±21.9) μmol/L vs (117.7±27.4) μmol/L,P ＜ 0.01] and UA [(303.5±66.9)μmol/L vs (371.092.4) μmol/L,P＜ 0.01].Compared to MDFR group,FDFR group showed lower GFR [(72.4±25.3) ml/min vs (82.7± 18.7) ml/min,P ＜ 0.05] 1 year after operation.Compared to MDMR group,FDFR group showed lower UA [(322.9±69.7) μmol/L vs (376.0±66.2) μmol/L,P ＜ 0.05] 2 years after operation.Compared to FDMR group,FDFR group showed lower Scr [(88.7 ±27.0) μmol/L vs (112.7±27.8) μmol/L,P ＜ 0.05] and UA [(318.3 ±61.2) μmol/L vs (396.2± 100.3) μmol/L,P ＜ 0.05] 3 years after operation.5 years after operation,there were no significant differences in above indexes,the incidence of slow graft function,acute rejection and survival of graft and patient among groups.Conclusions Male recipients of female donors have the worst renal function while female recipients have better outcomes after operation.

Subacute thyroiditis can cause destruction of thyroid follicles and subsequent transient thyrotoxicosis.In cases of simultaneous occurrences of subacute thyroiditis and Graves' disease,the former may be missed and thus may further exacerbate thyrotoxicosis.Herein,we report in detail a case with abrupt onset of thyrotoxic heart disease when taking anti-thyroid medications,in order to call attention to the diagnosis and treatment of concomitant Graves' disease and subacute thyroiditis.

[ ABSTRACT] AIM: To investigate the effect of miR-496 over-expression on the growth and metastasis of colon cancer cells and its molecular mechanism .METHODS:The proteins interacting with miR-496 were screened by bioinfor-matic method.The levels of miR-496, CTNNB1 mRNA and β-catenin protein in colon cancer cell lines , HT29, HCT116 and SW480, and normal colonic epithelial cell line NCM 460 were detected by real-time PCR and Western blot .HT29, HCT116 and SW480 cells were transfected with miR-496 mimics using Lipofectamine 2000 and named as HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimics cells, respectively, and the cells transfected with the scramble served as negative control .The cell viability, lactate dehydrogenase (LDH) leakage, and colony formation and metastatic abilities were determined by MTT assay , LDH assay, colony formation assay and Transwell method , respective-ly.The promoter activity of miR-496 was measured using luciferase reporter gene assay .The protein levels of β-catenin, eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), p-4E-BP1, low-density lipoprotein receptor-related protein 6(LRP6), p-LRP6, MMP-7, MMP-9, MMP-13 and TIMP-2 were monitored by Western blot.RESULTS:Endog-enous miR-406 interacted with β-catenin was found in the colon cancer cells .Low miR-496 expression in the HT29, HCT116 and SW480 cells and high miR-496 expression in NCM460 cells were detected.In contrast, high β-catenin ex-pression was found in the HT29, HCT116 and SW480 cells and low β-catenin expression was observed in the NCM460 cells.Compared with control group , the cell viability, colony formation rate and the number of metastatic cells remarkably decreased in the HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimic cells (P<0.05).The promoter activity of miR-496 was significantly increased in colon cancer cells transfected with miR-496 mimics, and was 1.75, 2.04 and 1.61 times as high as control group .miR-496 over-expression inhibited β-catenin levels, and p-4E-BP1 and p-LRP6 protein levels were also reduced .siRNA-or over-expressed miR-496-mediated β-catenin down-regulation in-hibited MMP-7 and MMP-9 expression , but promoted TIMP-2 expression .CONCLUSION:The expression level of miR-496 in the colon cancer cells is low , but in the normal colonic epithelial cells is high .miR-496 over-expression inhibits the protein levels of MMP-7 and MMP-9, and promotes the protein expression of TIMP-2 via inhibiting Wnt/β-catenin path-way, thus suppressing malignant phenotype in the colon cancer cells .

Objective To investigate the effects of paeonol on renin-angiotensin system (RAS) occurred on the development of ventricular remodeling after acute myocardial infarction (AMI) in rats. Methods The left anterior descending coronary artery was ligated to establish the model of AMI in male SD rats. Six groups were set up:sham-operation group, AMI model group, captopril control group, paeonol low dose group (6 mg/kg), paeonol middle dose group (9 mg/kg) and paeonol high dose group (12 mg/kg). Rats were given treatment for 4 weeks after the AMI model was established. HE staining was used to observe changes of myocardial tissue. Real-time PCR was used to detect the mRNA levels of angiotensinogen (AGT), angiotensin Ⅱ receptor type1(AGTR1) and endothelin (ET)-1 of six groups. Western blot assay was used to detect the protein levels of peptidyl-dipeptidase A (ACE), angiotensin Ⅱ(Ang)-Ⅱand AGTR1 in six groups. Results The transcription of AGT, AGTR1, ET-1mRNA and the expressions of ACE, Ang-Ⅱ and AGTR1 protein were significantly higher in myocardial tissue of AMI rats than those of sham-operation rats (P＜0.05). Compared with model group, the expressions of AGT, AGTR1, ET-1mRNA and ACE, Ang-Ⅱ, AGTR1 protein were significantly decreased in paeonol high dose group and captopril control group (P＜0.05). Paeonol reduced the expressions of those mRNA and protein levels in a significant dose dependent manner. Conclusion Paeonol can slow down the deterioration of the ventricular remodeling after AMI in rats, which may be related to the inhibition of over-activation of RAS.

Objective To investigate the expression levels of somatostatin (SST) gene in endometrial adenocarcinoma tissues and cell lines,and the effects of over-expression of SST gene on the proliferation and invasion of endometrial cancer cell line Ishikawa in vitro.Methods Tissue sections of normal endometrium,endometrioid adenocarcinoma and uterine papillary serous carcinoma were selected to detect the expressions of SST by immunohistochemical method.The total RNA was extracted from fresh specimens that were confirmed as endometrioid adenocarcinoma.According to FIGO staging,samples included G1 (7 cases),G2 (6 cases) and G3 (5 cases) of endometrioid adenocarcinoma.The SST expression levels were detected by real-time PCR.Three endometrial cancer cell lines,Ishikawa,HEC-1A and KLE,were selected and the expression levels of SST were detected by real-time PCR and Western blot assay.Transfection was performed with pLVX-SST and pLVX.The transfection efficiency was observed by fluorescence confocal microscopy.The protein levels of SST were detected by Western blot assay.The assays of CCK-8 and transwell were applied to examine variations in cell proliferation and invasion.Results Immunohistochemical results showed that SST expression was increased in endometrioid adenocarcinoma and uterine papillary serous carcinoma compared with that of normal endometrium.Real-time PCR showed that SST expression was significantly increased in G3 compared with that of G1 and G2 in endometrioid adenocarcinoma (P ＜ 0.05).No matter mRNA or protein,SST levels were significantly increased in endometrial cancer cell line KLE compared with those of Ishikawa and HEC-1A,and the expression levels of SST mRNA and protein were significantly increased in HEC-1A group than those of Ishikawa group (P＜0.05).The expression of SST protein was significantly higher in the group of Ish-SST after 2 generations compared with that of Ish-ctr group.There were no significant differences in cell proliferation and invasive ability after over-expression of SST between Ishikawa cell group and control group (P ＞ 0.05).Conclusion SST is highly expressed in poorly differentiated endometrial cancer cells.The proliferation and invasion are not increased after the over-expression of SST in Ishikawa cell line of endometrial cancer.

AIM: To evaluate the expression of fibroblast growth factor receptor 3 (FGFR3) in acute lymphocytic leukemia (ALL) patients and its contribution to the proliferation of circulating endothelial cells (CECs).METHODS: The mRNA expression levels of FGFR3 in 44 patients with ALL were assayed by RT-PCR.Overall survival (OS) rates of the patients in FGFR3+ group and FGFR3-group were estimated by Kaplan-Meier analysis.The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cell counts, antigen expression, growth curve and colony forming rate of the CECs in the 2 groups were determined.The FGFR3 expression of CECs was identified by immunofluorescence staining.RESULTS: The positive rate of FGFR3 mRNA expression was 43.2% in 44 ALL patients with normal karyotype.T-ALL expressed higher level of FGFR3 than B-ALL (P<0.05).FGFR3 was over-expressed in ALL patients with bone marrow blast proportion ≥5% (P<0.05).The probability of OS was significantly lower in FGFR3+ group than that in FGFR3-group (P<0.05).The sorted CECs highly expressed CD31, CD144, VEGFR-2 and CD146, and rarely expressed CD45.The counts of CECs and expression level of CD133 significantly increased in FGFR3+ group compared with FGFR3-group.The same result of the amount of colony formation was observed (P<0.05).There was significant difference at 3 time points of cultured CECs count in vitro between FGFR3+ group and FGFR3-group (P<0.05).The positive rate of FGFR3 expression of CECs from 19 ALL-FGFR3+ patients was (29.00±15.71)%.CONCLUSION: The over-expression of FGFR3 gene in ALL may be helpful to evaluate the prolife-ration of CECs, and become a double target with anti-tumor and anti-angiogenesis effects to offer more choice for molecular therapy in the future.

Severe Acute Respiratory Syndrome(SARS)is a deadly infectious disease caused by SARS Coronavirus(SARS-CoV).Inactivated SARS-CoV has been explored as a vaccine against SARS-CoV.However,safe and potent adjuvants,especially with more efficient and economical needle-free vaccination are always needed more urgently in a pandemic.The development of a safe and effective mucosal adjuvant and vaccine for prevention of emergent infectious diseases such as SARS will be an important advancement.PIKA,a stabilized derivative of Poly(I:C),was previously reported to be safe and potent as adjuvant in mouse models.In the present study,we demonstrated that the intraperitoneal and intranasal co-administration of inactivated SARS-CoV vaccine together with this improved Poly(I:C)derivative induced strong anti-SARS-CoV mucosal and systemic humoral immune responses with neutralizing activity against pseudotyped virus.Although intraperitoneal immunization of inactivated SARS-CoV vaccine alone could induce a certain level of neutralizing activity in serum as well as in mucosal sites,co-administration of inactivated SARS-CoV vaccine with PIKA as adjuvant could induce a much higher neutralizing activity.When intranasal immunization was used,PIKA was obligatorily for inducing neutralizing activity in serum as well as in mucosal sites and was correlated with both mucosal IgA and mucosal IgG response.Overall,PIKA could be a good mucosal adjuvant candidate for inactivated SARS-CoV vaccine for use in possible future pandemic.