The influence of short hairpin RNA (shRNA)-mediated osteopontin (OPN) gene silencing on the proliferation and invasion of human renal cancer ACHN cells was investigated. Four types of OPN shRNA recombinant plasmids were constructed and RT-PCR assays were used to screen the most highly functional shRNA recombinant plasmids, which were transferred into the cultured ACHN cells by Lipofectamine 2000. The cells transfected by shRNA expression vectors (ACHN/OPN) were visualized under an inverted microscope and screened by G418. Untreated cells (ACHN) and cells transfected by mock vectors (ACHN/Vect) were used as control groups. The expression levels of OPN mRNA and protein were detected by real-time PCR and Western blot respectively. The cell cycle and ratios of apoptotic cells were assessed by flow cytometry. MTT method was used for drawing the growth curve and observing cell proliferation in vitro. The abilities of migration and invasion in three groups were measured by Transwell chamber test. The expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 in three groups were examined by Western blot. Our results showed that the recombinant plasmid could be successfully transferred into ACHN cells by LipofectamineTM 2000. Compared with untreated cells, the expression levels of OPN mRNA and protein in ACHN/OPN cells were decreased by 59.68% and 76.42%, respectively (P<0.05), ACHN/OPN cells were blocked in S phase and apoptotic ratio increased significantly (P<0.05), however, no significant differences were found between ACHN/Vect and ACHN. Recombinant plasmid significantly attenuated expression levels of MMP-2 and MMP-9 proteins and suppressed the proliferation, migration, and invasion of ACHN cells. This study suggested that OPN may play an important role in the growth and invasion of human renal cancer ACHN cells, and these processes are correlated with the activations of MMP-2 and MMP-9. Our data provided preliminary experimental evidence for the feasibility of RNA interference technology in gene therapy of human renal cancer.

Objective To investigate the characteristic sonographic and pathological features of breast ductal carcinoma in situ (DCIS) without microcalcifications on mammography (MG).Methods Forty cases of DCIS without microcalcifications on MG were retrospectively reviewed.The 40 lesions were classified into mass and non-mass groups according to their sonographic findings.The pathological subtypes and nuclear grades of these cases were also analyzed.Fisher exact test was used to compare the differences of the sonographic accuracy rate,sonographic microcalcification rate,pathological nuclear grade and subtype rate between mass and non-mass groups.Results No abnormal finding was found in sixteen cases (40.0％)on MG and only one case (2.5％) on ultrasonography (US),respectively.The most common sonographic feature of DCIS without microcalcifications on MG were masses (75.0％,30/40),and other sonographic findings were round/oval and irregular shape,microlobulated margin,heterogeneous hypoechogenicity and isoechogenicity,and posterior acoustic feature.Ductal dilatations and heterogeneous isoechogenicity were present in most non-mass lesions of DCIS without microcalcifications on MG (22.5％,9/40).The ultrasonographic microcalcifications were found in 5 cases of DCIS without microcalcifications on MG.The common pathological features of DCIS without microcalcifications on MG were medium-low nuclear grade (85.0％,34/40) and noncomedo (87.5％,35/40).The difference of US accuracy rate in mass and non-mass groups was statistically significant [73.3％ (22/30) vs 33.3％ (3/9),P=0.047].The differences of US microcalcification rate,pathological subtype and nuclear grade were not significant (P=1.000,0.070).Conclusions The mass appearance and medium-low nuclear grade were most common sonographic findings and pathological features of DCIS without microcalcifications on MG.Ultrasonography should be an helpful tool for improving the diagnostic sensitivity ofmammography in breast DCIS.

To design and synthesize a series of novel scutellarein 4'-L-amino acid prodrugs with more potent anti-oxidative activity and improved physicochemical properties. Scutellarein was used as lead compound, according to successful experience of improving bioavailability of oral administration drugs by active transport mechanism, principle of hybridization was used to introducing L-amino acid structural fragments at 4'-position of scutellarein to design and synthesize target scutellarein 4'-L-amino acid prodrugs. The synthetic compounds were tested on their physicochemical properties and in vitro anti-oxidative activity against H202 induced oxidative damage in PC12 cells. Five compounds were found to have more potent anti-oxidative activity than positive control VE. Moreover the physicochemical properties of synthesized compounds were evaluated, and the results revealed that L-amino acid ether derivatives are more stable (t1/2 9-92 h) than their corresponding ester derivatives (t1/2 0.5 h). Water solubility of scutellarein 4'-L-amino acid ester and ether derivatives were 1 796-4 100 microg.mL-1 and 27.7-81.1 microg.mL-1 respectively, in comparison with scutellarin, the solubility of compounds 18, 19 and 22, 24-27 increased about 120-280 fold and 2-6 fold respectively. All these results suggested that L-amino acid prodrug strategy has significant potential in scutellarein prodrug design.

The roles of Indian hedgehog (Ihh) signaling pathway in the proliferation and apoptosis of precartilaginous stem cells (PSCs) were investigated.PSCs,labeled with fibroblast growth factor receptor 3 (FGFR-3),were isolated from neonatal rats by immanomagnetic separation.After identifi-cation with FGFR-3 and Col Ⅱ,the cells were incubated with different concentrations of cyclopamine (cyclo),the specific inhibitor of lhh signaling pathway.The morphologic changes of the cells were observed under the inverted phase contrast microscope.The mRNA expression levels of Ibh,para-thyroid hormonerelated peptide (PTHrP),protein Patched (Ptch),Bcl-2 and p21 were detected by RT-PCR.The protein expression levels of Ihh and Ptch were measured by Western blot.MTT assay was used to examine the effects of cyclo on proliferation of PSCs.Apoptosis rate of PSCs was exam-ined by Annexin V/PI assay of flow cytometric analyses.After PSCs were incubated with cyclo,ob-vious morphologic changes were observed as compared with the control group.The mRNA expres-sion levels of PTHrP,Ptch and Bcl-2 were decreased to varying degrees in a cyclo dose-dependent manner.However,the expression levels of lhh and p21 mRNA were increased.The protein expres-sion of Ptch and Ihh had the same change as the mRNA expression.Meanwhile,cyclo could obvi-ously inhibit the proliferation and promote the apoptosis of PSCs.The results indicated that Ihh sig-naling pathway plays an important role in regulating the proliferation and apoptosis of PSCs,which is probably mediated by Bcl-2 and p21.

The influence of short hairpin RNA(shRNA)-mediated osteopontin(OPN)gene silencing on the proliferation and invasion of human renal cancer ACHN cells was investigated.Four types of OPN shRNA recombinant plasmids were constructed and RT-PCR assays were used to screen the most highly functional shRNA recombinant plasmids,which were transferred into the cultured ACHN cells by LipofectamineTM 2000.The cells transfected by shRNA expression vectors(ACHN/OPN)were visualized under an inverted microscope and screened by G418.Untreated cells(ACHN)and cells transfected by mock vectors(ACHN/Vect)were used as control groups.The expression levels of OPN mRNA and protein were detected by real-time PCR and Western blot respectively.The cell cycle and ratios of apoptotic cells were assessed by flow cytometry.MTT method was used for drawing the growth curve and observing cell proliferation in vitro.The abilities of migration and invasion in three groups were measured by Transwell chamber test.The expression levels of matrix metalloproteinase(MMP)-2 and MMP-9 in three groups were examined by Western blot.Our results showed that the recombinant plasmid could be successfully transferred into ACHN cells by LipofectamineTM 2000.Compared with untreated cells,the expression levels of OPN mRNA and protein in ACHN/OPN cells were decreased by59.68% and 76.42%,respectively(P<0.05),ACHN/OPN cells were blocked in S phase and apoptotic ratio increased significantly(P<0.05),however,no significant differences were found between ACHN/Vect and ACHN.Recombinant plasmid significantly attenuated expression levels of MMP-2 and MMP-9 proteins and suppressed the proliferation,migration,and invasion of ACHN cells.This study suggested that OPN may play an important role in the growth and invasion of human renal cancer ACHN cells,and these processes are correlated with the activations of MMP-2 and MMP-9.Our data provided preliminary experimental evidence for the feasibility of RNA interference technology in gene therapy of human renal cancer.

Objective:To investigate the clinical features and influencing factors of early-onset gout.Methods:Male patients with gout admitted to Department of Endocrinology and Metabolism were recruited from 2015 to 2018. Patients with gout onset before age 30 were defined as the " early-onset" group, and those with onset at 30~60 years were defined as the "late-onset" group. Clinical characteristics were compared between two groups. Factors associated with early-onset gout were analyzed.Results:A total of 1 243 male patients were enrolled in this study; 480 individuals were in the early-onset, and 763 in the late-onset groups. Compared with the late-onset group, patients with early-onset gout had higher consumption rates of sugar-sweetened beverage(28.0% vs 15.0%, P=0.001), a higher homeostasis model assessment for insulin resistance level(3.78±2.93 vs 3.10±2.39, P<0.01), and larger proportions of family histories of diabetes(30.8% vs 20.4%, P<0.01)and hypertension(51.2% vs 42.6%, P=0.003). Logistic regression analysis showed that factors associated with early-onset gout were drinking sugar-sweetened beverage( P=0.012), family history of diabetes( P=0.037). Conclusion:Early-onset gout was associated with a family history of diabetes. Patients with family histories of diabetes are more likely to have early-onset gout, which may be associated with a common genetic basis.

Objective To investigate the correlation between plasma inflammatory factors [IL-1β, IL-6, IL-10, IL-12, IL-17, IL-23, TNF-α, TGF-β, IFN-γ, C-reactive protein (CPR) CCL-5] and hand-foot syndrome in colorectal cancer patients after taking capecitabine. Methods 35 colorectal cancer patients treated with capecitabine were collected and the degree of severity was divided according to the hand-foot syndrome grading diagnostic criteria. The concentrations of inflammatory factors in plasma were determined by ELISA kits. Results The standard curve of all inflammatory cytokines were linear (r>0.9900), and plasma concentrations of inflammatory cytokines in patients with colorectal cancer were determined. The concentration of TNF-α changed obviously, which had reference value. Conclusion The concentrations of different inflammatory factors were different and the concentration of TNF-α was closely correlated with the severity of hand-foot syndrome.