One hundred and forty-two patients on maintenance hemodialysis were enrolled in the study.According to the guideline of American Society of Echocardiography 97 patients (68.3％) were classified as pulmonary artery hypertension [PAH,pulmonary artery systolic pressure (PASP) ＞ 35 mmHg (1 mmHg =0.133 kPa)] and 45 patients (31.7％) as non-PAH (PASP ≤ 35 mmHg).High sensitivity Creactive protein (hs-CRP) and tumor necrosis factor-α (TNF-α) were measured by automatic analyzer and enzyme linked immunosorbent assay (ELISA),respectively.There were significant differences in CRP and TNF-α levels between PAH and non-PAH patients (P ＜ 0.05).Multivariate linear regression showed that TNF-α and interdialytic weight gain were positively correlated with PAH and left ventricular ejection fraction was negatively correlated with PAH (P ＜ 0.01).In the multivariate Cox proportional hazards models,PAH and TNF-α level were independently associated with higher risk for all-cause death (HR =1.06,95％ CI:1.03-1.09 and HR =1.31,95％ CI:1.17-1.46,respectively).In Kaplan-Meier survival analysis,the risk of all-cause mortality increased in parallel with PASP.The risk of death in patients with PASP ＞45 mmHg was higher than that in patients with PASP ≤45 mmHg (log-rank test:x2 =6.58,P =0.010),the risk was 2.82-fold (HR =2.82,95％ CI:1.38-5.77,P =0.004).

Objective:To investigate the effects of miR-125b on radiosensitivity of cervical cancer cells and its possible downstream mechanism.Methods:The expression of miR-125b and Foxp3 in cervical cancer tissues and cells was detected by RT-qPCR. The HeLa cells were irradiated with 0, 2, 4 and 6 Gy of X-rays. The expression of miR-125b and Foxp3 in each group was detected by RT-qPCR. After downregulation of miR-125b expression and 6 Gy X-ray irradiation, the proliferation ability of HeLa cells was detected by MTT assay, and the expression of Bax and Bcl-2 proteins were detected by Western blot. The relationship between miR-125b and Foxp3 was detected by Targetscan and Dual luciferin reporter assay. After downregulation of Foxp3 expression and 6 Gy X-ray irradiation, the proliferation ability of HeLa cells was detected by MTT assay, and the expression of Bax and Bcl-2 proteins were detected by Western blot. The effects of miR-125b on radiosensitivity of HeLa cells through Foxp3 were detected. After down-regulation of Foxp3, the contents of IL-10 and TGF-β in supernatant were detected by ELISA.Results:The expression of miR-125b in the tissues and cells of cervical cancer was significantly decreased, while the expression of Foxp3 was significantly increased. The expression of miR-125b in HeLa cells was increased after radiation in a dose dependent manner. The expression of Foxp3 in HeLa cells was decreased after radiation in a dose dependent manner. After 6 Gy X-ray irradiation of HeLa cells, down-regulation of miR-125b increased the cell proliferation capacity, significantly reduced the expression of Bax and increased the expression of Bcl-2. miR-125b targets Foxp3 and negatively regulates Foxp3 expression. After 6 Gy X-ray irradiation of HeLa cells, down-regulation of Foxp3 significantly reduced the proliferation capacity of HeLa cells, increased the expression of Bax and decreased the expression of Bcl-2. Overexpression of miR-125b can enhance radiosensitivity of HeLa cells through Foxp3.After 6 Gy X-ray irradiation, down-regulation of Foxp3 reduced the expression of IL-10 and TGF-β in cells.Conclusions:Upregulation of miR-125b enhances the radiosensitivity of cervical cancer cells by targeting and negatively regulating Foxp3, and the mechanism of that may be related to the down-regulation of Foxp3 to reduce the expression of IL-10 and TGF-β in the cells.

Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVE:To investigate the effects of calcium dobesilate on the ultrastructure of glomerular basement membrane(GBM)in diabetic rats.METHODS:30rats underwent unilateral nephrectomy,of whom,10were assigned to the normal control group(group NC),another20were induced to diabetic models(DM)by intraperitoneal injection of1%STZ,the models which have finished were divided into DM control group and calcium dobesilate group(group DD),each group admin?istered with distilled water and calcium dobesilate respectively,12weeks later,the change in the ultrastructure of kidneys and endogenous creatinine clearance rate(CrCl)in each group were observed.RESULTS:After12weeks,endogenous CrCl in group DD was significantly higher than that in group DM;Electron microscope showed that thickening of glomerular capillary basement membrane in group DD was less than that in group DM.CONCLUSION:Calcium dobesilate could improve the ul?trastructure in kidney of diabetic rats,prevent GBM thickening,and protect filtration barrier of renal glomerulus.

Objective To identify new microsatellite loci from genome sequence database for the study of poly-morphicsm of Schistosoma japonicum. Methods Schistosoma japonicum isolates were obtained from seven endemic sites in China: Tongling and Guichi counties of Anhui Province, Duchang county of Jiangxi Province, Changde and Yueyang Cities of Hunan Province, Shashi City of Hubei Province, Xichang City of Sichuan Province. In order to study the genetic variance, genomic DNAs of 96 individual adult worms were screened against 17 new Schistosoma japonicum microsatellites and the raw data were analyzed by GenMapper 4.0. Furthermore, the varieties of alleles were inverstigated using GenAlEx 6 and genetic distances within a subpopulation (GenClone) and among populations(UPGMA, MEGA 3.1) were analyzed. Results High levels of polymorphism were found between and within population samples, and significant genetic diversity was observed among the seven subpopulations.Within Jiangxi population, most genetic distances (17 loci) among samples range from 25 to 32, indicating a significant genetic diversity. There are three clusters among the seven populations: Jiangxi, Tonglin, Shashi and Changde population, with the genetics distances ranging from 0.017 8 to 0.036 3; Guichi and Yueyang population belong to another cluster, with the genetic distance of 0.024 7; However, Xichang population is an unique group. Its genetic distances to other populations are notable with a range from 0.019 2 to 0.069 3. Conclusion The 17 new polymorphic microsatellites identified may be used as suitable markers for the study on population genetics of Schistosoma japonicum and the genetic variance of the worms seems to be complicated.

OBJECTIVETo assess the accuracy of ultrasound-guided 16G and 18G core needle biopsy for detecting ultrasound visible breast lesions with different sonographic features.

METHODSA total of 955 sonographically detected breast lesions examined with ultrasound-guided core needle biopsy (US-CNB) and subsequently surgically excised from July 2005 to July 2012 were retrospectively reviewed. Histological findings of US-CNB and the surgical specimens were analyzed for agreements, sensitivities, false negative rates, and underestimate rates according to different sonographic features.

RESULTSThe pathological results of the US-CNB showed malignant lesions in 84.1%, high-risk lesions in 8.4%, and benign lesions in 7.5% of the samples. The overall agreement rates were 92.4% for 16G CNB and 92.8% for 18G CNB; their complete sensitivities and false negative rates were both 98.6% and 1.4%, respectively; the high-risk underestimate rates and DCIS underestimate rates were 48.0% and 46.2% for 16G CNB vs 53.3% and 41.2% for 18G CNB, showing no significant difference between the two groups (P>0.01). For both 16G and 18G CNB, the agreements were better for mass lesions than for non-mass lesions (P<0.01). For the mass lesions with a diameter no greater than 10 mm, the agreement rates were lower than the overall data (P<0.01). Calcification in the lesions did not affect the agreement rates (P>0.01).

CONCLUSIONUltrasound-guided 16G and 18G CNB are both accurate methods for evaluating ultrasound visible breast mass lesions with a diameter larger than 10 mm.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; instrumentation ; methods ; Biopsy, Needle ; instrumentation ; methods ; Breast ; pathology ; Female ; Humans ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography, Mammary ; Young Adult

To observe the effects of calcium dobesilate on the expression of glomerular tissue inhibitor of metalloproteinase 1 (TIMP1), collagen Ⅳ, and ultrastructure of glomerular basement membrane in diabetic rats, rats model of diabetes was established by unilateral nephrectomy and intraperitoneal injection of 1% STZ (55 mg/kg), and rats were administered calcium dobesilate 100 mg/kg (DD group) or distilled water (DM group) respectively. 12 weeks later, the changes in the renal ultrastructure and creatinine clearance rate (Ccr) were examined in each group. The expression of glomerular TIMP1 and collagen Ⅳ were studied by immunohistochemical staining. Our results showed that after 12 weeks, the Ccr in DD group increased and was significantly higher than that in DM group. Electron microscopy showed that thickness of glomerular capillary basement membrane (GBM) in Group DD was less than that of DM group. No hyperplasia of collagen fibers was found, and the distance between the holes of endothelial cells in DD group was not as even as that in the normal group, but more even than that of DM group, and podocyte processes was still in order. Immunohistochemical staining of glomeruli showed that expression of TIMP1 and collagen Ⅳ in DD group were significantly less than those of DM group DM. It is concluded that calcium dobesilate can improve diabetic nephropathy by inhibiting the over- accumulation of collagen Ⅳ and calcium dobesilate may also contribute to diabetes by inhibiting the expression of TIMP1.

Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of Ca²⁺-calmodulin using the activities of 20 kDa myosin light chain (MLC₂₀) phosphorylation and myosin Mg²⁺-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low Ca²⁺, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high Ca²⁺, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of MLC₂₀ and Mg²⁺-ATPase activities of Ca²⁺- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.
Animals ; Ardisia ; Atropine ; Blotting, Western ; Epinephrine ; Gastric Emptying ; Gastrointestinal Motility* ; Histamine ; In Vitro Techniques ; Muscle, Smooth* ; Myosin Light Chains* ; Myosin-Light-Chain Kinase* ; Myosins* ; Phosphorylation* ; Rats ; Saponins

Objective:To investigate the regulatory mechanism of long-chain non-coding RNA (lncRNA) MEG3 on the sensitivity of lung cancer cell line H1299 to irradiation.Methods:The expression of MEG3 and miR-21-5p in lung cancer cell line H1299 was detected by qRT-PCR. Overexpression control group (transfected with pcDNA3.1), MEG3 overexpression group (transfected with pcDNA3.1-MEG3), miR-NC inhibition group (transfected anti-miR-NC), miR-21-5p inhibition group (transfected with anti-miR-21-5p), MEG3 overexpression+ miR-NC overexpression group (co-transfected with pcDNA3.1-MEG3 and miR-NC), MEG3 overexpression+ miR-21-5p overexpression group (co-transfected with pcDNA3.1-MEG3 and miR-21-5p mimics) were all transfected into H1299 cells by liposome method treated with 4 Gy irradiation. Cell survival fraction was detected by colony formation assay. Cell apoptosis was detected by flow cytometry. The binding of MEG3 to miR-21-5p in cells was assessed by dual luciferase reporter assay.Results:Compared with normal lung epithelial cells, the expression of MEG3 was significantly decreased, whereas the expression of miR-21-5p was significantly increased in the radioresistant lung cancer cells H1299. Overexpression of MEG3 or inhibition of miR-21-5p could promote the apoptosis and enhance the radiosensitivity of H1299 cells. MEG3 could targetedly regulate the expression of miR-21-5p. Overexpression of miR-21-5p could reverse the enhanced radiosensitivity of MEG3 to H1299 cells.Conclusion:LncRNA MEG3 can enhance the sensitivity of lung cancer cells H1299 to irradiation. The mechanism may be related to targeting miR-21-5p.

Objective To assess the accuracy of ultrasound-guided 16G and 18G core needle biopsy for detecting ultrasound visible breast lesions with different sonographic features. Methods A total of 955 sonographically detected breast lesions examined with ultrasound-guided core needle biopsy (US-CNB) and subsequently surgically excised from July 2005 to July 2012 were retrospectively reviewed. Histological findings of US- CNB and the surgical specimens were analyzed for agreements, sensitivities, false negative rates, and underestimate rates according to different sonographic features. Results The pathological results of the US-CNB showed malignant lesions in 84.1%, high-risk lesions in 8.4%, and benign lesions in 7.5%of the samples. The overall agreement rates were 92.4% for 16G CNB and 92.8% for 18G CNB; their complete sensitivities and false negative rates were both 98.6%and 1.4%, respectively;the high-risk underestimate rates and DCIS underestimate rates were 48.0% and 46.2% for 16G CNB vs 53.3% and 41.2% for 18G CNB, showing no significant difference between the two groups (P>0.01). For both 16G and 18G CNB, the agreements were better for mass lesions than for non-mass lesions (P<0.01). For the mass lesions with a diameter no greater than 10 mm, the agreement rates were lower than the overall data (P<0.01). Calcification in the lesions did not affect the agreement rates (P>0.01). Conclusion Ultrasound-guided 16G and 18G CNB are both accurate methods for evaluating ultrasound visible breast mass lesions with a diameter larger than 10 mm.