Objective To estimate the value of transrectal ultrasound/magnetic resonance imaging (TRUS/MR) fusion targeted prostate biopsy(targeted biopsy,TB) in the biopsy naive patients.Methods Between September 2015 and September 2016,91 patients with PI-RADS ≥ 3 suspicious regions on the multiparametric magnetic resonance imaging (mpMRI) were retrospectively evaluated.The age of patients was 46-83 years (median 68).Serum PSA level before biopsy was 1.2-85 ng/ml (median 11.2 ng/ ml),in which 36 cases with PSA ＜ 10 ng/ml,30 cases 10-20 ng/ml,and 25 cases ＞ 20 ng/ml.Two-core TB using real-time virtual sonography (RVS) platform for mpMRI-suspicious lesions was followed by 12-core systematic biopsy (SB).The detection rates for any cancer (PCa) and clinically significant prostate cancer (CsPCa) were compared between TB and SB.Results The total detection rate for PCa was 57.1％,with a comparable positive rate between TB (44.0％) and SB (51.7％) groups which did not significantly differ (P =0.14).The proportion of CsPCa in TB group was higher than that in SB group (80.0％ vs.68.1％,P =0.21).In TB group,detection of PCa for grade 5 lesions was significantly higher than that for grade 3 lesions (77.1％ vs.10.3％,P ＜0.001).Detection of PCa was comparable between TB and SB groups in different regions of PSA ＜ 10 ng/ml,10 ～ 20ng/ml and ＞ 20ng/ml (27.8％ vs.36.1％,50％ vs.56.7％,60％ vs.68％,respectively).Conclusions This study revealed a similar rate of prostate cancer detection between 2-core targeted biopsy guided by TRUS/MR fusion and 12-core random biopsy in different PSA regions for no prior biopsy men.TB maybe tend to detect high proportion of CsPCa.PI-RADS is instructive to select appropriate patients for TB.

Objective To evaluate the feasibility and efficacy of intra-cavity contrast enhanced ultrasound (ICCE-US) in percutanous nephrolithotomy for nephrolithiasis patients with slight or no hydronephrosis.Methods From March 2016 to March 2017,ICCE-US-guided PCNL was performed in 35,patients who had kidney stones with slight hydronephrosis in 11 and without hydronephrosis in 24.The sample comprised 20 males and 15 females,including 10 with renal pelvic calculi alone,17 with renal pelvic calculi combined with renal calyx calculi,and 8 with partial staghorn calculi.Mean age was 46.8 years (ranging,28-75 years).The size of calculi ranged from 2.5 cm to 5.0 cm [mean(3.6 ± 1.2) cm].In the prone position,the preferred calyces are the posterior ones,which were enhanced by sulfur hexafluoride microbubbles (SonoVue) retrogradely injected through ureteral catheter.An 18-gauge needle was inserted toward the desirable calyx.Successful renal entry was confirmed by administration of ultrasound contrast agents into the collecting system via the needle regardless of whether spontaneous urine drainage was observed.A guidewire was passed through the needle to renal collecting system.Subsequently,the needle was removed.And the renal tract was dilated to F18-F20 size with dilators.Finally,holmium laser lithotripsy was performed through nephroscopy.Results Posterior calyces and its fornix were revealed under contrast-enhanced ultrasound in all patients.The successive access rate was 100％.The average time for establishing the access was (8 ±2.6)min (ranging 5-10 min).The mean number of needle passes was (1.5 ± 0.3) times per kidney,ranged from 1 to 3 times.Hemoglobin level averagely reduced (11.3 ± 3.7) g/ L (ranging 6-15 g/L) within 24 hours postoperatively.No major complications,such as adjacent organs injuries or collecting system perforation were observed.No blood transfusion was needed.The mean hospital stay was (5.6 ± 1.2) days (ranging 5-7 days).The stone-free rate was 91.4％ (32/35).Conclusions ICCE-US can demonstrate clearly about the posterior calyces and its fornix as well as puncture needle site by contrast enhancement in the nephrolithiasis patients with slight or no hydronephrosis.It has the potential to improve the accuracy and successive rate of puncture resulting in a decrease in the puncture-related complications.

Objective To evaluate the bacteriostatic effect of recombinant human lactoferrin(rhLF) on Helicobacter(H .) py‐lori and its influence on CagA ,Ure and gastric mucosal IL‐8 .Methods The minimum inhibitory concentration(MIC)and the influ‐ence of different drug concentrations on the proliferation of H .pylori were detected .The effects of rhLF on the mRNA and protein expressions of CagA and Ure in H .pylori were detected by RT‐PCR and Western blot ,respectively .The animal study :Balb/c mice were adopted and assigned randomly into four groups ,including the standard triple+rhLF(group A) ,rhLF(group B) ,standard tri‐ple(group C) and normal saline(group D) .The histopathological HE staining was used to observe the gastric inflammation and ELISA was used to detect the IL‐8 level of gastric tissue in each group .Results MIC was 0 .5 mg/mL ,moreover rhLF inhibited the bacterial growth and proliferation with a concentration‐dependent manner .rhLF could reduce the expression of H .pylori major viru‐lence factor CagA ,mRNA and protein of Ure .Comparing the group A with the group B ,C and D ,the gastric mucosal inflammation score and the IL‐8 levels of gastric tissue homogenates had statistically significant differences(P<0 .05) .Conclusion rhLF inhibits the growth and proliferation of H .pylori ,moreover inhibit the expression of major virulence factor CagA in H .pylori ,mRNA and protein of Ure in different degrees ,weakens its pathogenicity ,meanwhile reduces the IL‐8 level in mice gastric mucosa ,and allevi‐ates H .pylori related gastric mucosal inflammatory response .

OBJECTIVE To investigate the effect of clopidogrel(Clog),a platelet aggregation inhibitor,on the development of colitis-associated colon cancer(CAC)and its possible mechanism. METHODS To establish a CAC model,male BALB/c mice were treated with single azoxymethane(AOM) 10 mg · kg-1 by ip. One week later,the mice drank 2.5% dextran sulfate sodium(DSS)for one week and water for two weeks,which lasted three cycles. From the first day mice received 2.5%DSS water, Clog 12.5,25.0 and 50.0 mg · kg-1 was ig administered once a day. Body mass,clinical symptoms,the number of colon tumor and tumor size in colon tissue were recorded. Hyperplasia of tumors was analyzed by HE staining. In the early inflammatory phase of the CAC model,the length of colons was measured, histological structure and epithelium cell proliferation of colon tissues were evaluated by HE staining and Ki67 staining,respectively. In the tumorigenesis and progression phase of the CAC model,epithe?lium cell proliferation of colon tissues was evaluated by Ki67 staining. The mRNA expression of tumor necrosis factor-α(TNF-α)was detected by real-time quantitative PCR. The expression of chemokine(C-X-C motif)ligand 2(CXCL2)and its receptor 2(CXCR2)in colon tissues was detected by PCR and immu?nohistochemistry. RESULTS Compared with model group,clinical symptoms of mice in Clog 12.5 mg · kg-1 group were alleviated,the size of colon tumors was decreased(P<0.05),and hyperplasia of tumors was reduced(P<0.05). During the inflammatory phase,the clinical symptoms of mice in Clog 12.5 mg·kg-1 group were significantly alleviated(P<0.05),the decrease of body mass was reduced(P<0.01),the colon shrinkage was ameliorated(P<0.01),the inflammatory injury and epithelium cell proliferation in colon tissues were reduced(P<0.05). During the tumorigenesis and progression phase,epithelium cell prolif?eration in colon tissues in Clog 12.5 mg·kg-1 group was reduced(P<0.01),and the mRNA and protein expression of TNF-α,CXCL2 and CXCR2 of colon tissues was decreased(P<0.05). CONCLUSION Clog can alleviate inflammation during the CAC early inflammatory phase and inhibit the formation of CAC. The antitumor effect of Clog may be related to the decrease in expression of CXCL2 and CXCR2.

Objective To explore the feasibility of adding a flexible linker between two-pore-domain potassium channel TREK-1 (TWIK related K + channel 1)monomers to construct a tandem-linked dimer.Methods PCR was used to add a flexible linker between the two TREK-1 monomers.The cRNA obtained from in vitro transcription using the above vector was injected into Xenopus oocytes.After 24 -48 h,currents were recorded from these oocytes using a two-electrode voltage clamp.The effects of extracellular Ba2 + and pH on TdTREK-1 were observed and compared with those of native dimeric TREK-1.Results The tandem-linked dimeric TdTREK-1 was highly expressed in Xenopus oocytes.The currents through these channels were inhibited by extracellular Ba2 +and acidification.Furthermore,the responsiveness of the concatenated dimers to these extracellular stimuli was similar to that of native dimers.Conclusion Adding a flexible linker between the two monomers to construct the tandem-linked dimer does not affect the expression and gating properties of TREK-1, suggesting that the method be feasible.Such a method will allow the manipulation of a single subunit,which will help basis study the structure and function of TREK-1.

Aim ToconstructHEK293cellsstablyex-pressing corticotropin releasing factor receptor 1 ( CRFR1 ) , and evaluate its function by the cAMP as-say.Methods CulturedHEK293cellsweretransfect-ed with CRFR1-expressing vector by Lipofectamine 2000 and were selected by using G418 . CRFR1 ex-pression was detected by Western blot, RT-PCR and immunofluorescence.Results Westernblot,RT-PCR and immunofluorescence data revealed that the HEK293 cells expressed CRFR1 protein stably. The dose-responsive relationship experiment revealed that CRF induced a CRFR1-mediated cAMP production in HEK293 cells with EC50 =(5. 64 ± 0. 05) × 10 -10 mol ·L-1.Conclusion HEK293celllinesstablyex-pressing CRFR1 were constructed successfully, which would provide a cellular model to facilitate the research on the biological function of CRFR1 and CRFR1-targe-ted drug screening.

In producing transgenic livestock, selectable marker genes (SMGs) are usually used to screen transgenic cells from numerous normal cells. That results in SMGs integrating into the genome and transmitting to offspring. In fact, SMGs could dramatically affect gene regulation at integration sites and also make the safety evaluation of transgenic animals complicated. In order to determine the deletion time and methods in the process of producing transgenic goats, the feasibility of deleting SMGs was explored by Cre/LoxP before or after somatic cell cloning. In addition, we compared the efficiency of protein transduction with plasmids co-transduction. We could delete 43.9% SMGs after screening out the transgenic cell clones, but these cells could not be applied to somatic cells cloning because of serious aging after two gene modifications. The SMG-free cells suitable for nuclear transfer were accessible by using the cells of transgenic goats, but this approach was more time consuming. Finally, we found that the Cre plasmid could delete SMGs with an efficiency of 7.81%, but about 30% in SMG-free cells had sequences of Cre plasmid. Compared with Cre plasmid, the integration of new exogenous gene could be avoided by TAT-CRE protein transduction, and the deletion rate of TAT-CRE transduction was between 43.9 and 72.8%. Therefore, TAT-Cre transduction could be an effective method for deleting selectable marker genes.
Animals ; Animals, Genetically Modified ; genetics ; Cloning, Organism ; veterinary ; Gene Knockout Techniques ; Gene Targeting ; methods ; Genes, Reporter ; Genetic Engineering ; Genetic Vectors ; genetics ; Goats ; genetics ; Integrases ; chemistry ; metabolism ; Recombination, Genetic ; Transgenes ; genetics

Antitumor effects of erythromycin and the related mechanism were investigated in the present study. Neuroblastoma cells (SH-SY5Y) were exposed to erythromycin at different concentrations for different durations. Cell proliferation was measured by cell counting, and cell viability was examined by MTT assay. Cell cycle phase distribution and the cytosolic calcium level were detected by flow cytometry. Mitochondrial membrane potential was measured by the JC-1 probe staining and fluorescent microscopy. The expression of an oncogene (c-Myc) and a tumor suppressor [p21 (WAF1/Cip1)] proteins was analyzed by using Western blotting. Erythromycin could inhibit the proliferation of SH-SY5Y cells in a concentration- and time-dependent manner. The cell cycle was arrested at S phase. Mitochondrial membrane potential collapsed and the cytosolic calcium was overloaded in SH-SY5Y cells when treated with erythromycin. The expression of c-Myc protein was down-regulated, while that of p21 (WAF1/Cip1) protein was up-regulated. It was concluded that erythromycin could restrain the proliferation of SH-SY5Y cells. The antitumor mechanism of erythromycin might involve regulating the expression of c-Myc and p21 (WAF1/Cip1) proteins.

Antitumor effects of erythromycin and the related mechanism were investigated in the present study.Neuroblastoma cells (SH-SY5Y) were exposed to erythromycin at different concentrations for different durations.Cell proliferation was measured by cell counting,and cell viability was examined by MTT assay.Cell cycle phase distribution and the cytosolic calcium level were detected by flow cytometry.Mitochondrial membrane potential was measured by the JC-1 probe staining and fluorescent microscopy.The expression of an oncogene (c-Myc) and a tumor suppressor [p21 (WAF1/Cipl)] proteins was analyzed by using Western blotting.Erythromycin could inhibit the proliferation of SH-SY5Y cells in a concentration- and time-dependent manner.The cell cycle was arrested at S phase.Mitochon drial membrane potential collapsed and the cytosolic calcium was overloaded in SH-SY5Y cells when treated with erythromycin.The expression of c-Myc protein was down-regulated,while that of p21 (WAF1/Cip1) protein was up-regulated.It was concluded that erythromycin could restrain the proliferation of SH-SY5Y cells.The antitumor mechanism of erythromycin might involve regulating the expression ofc-Myc and p21 (WAF1/Cip1) proteins.

Objective To establish a treatment proposal of thyroid adenoma by using percutaneous radiofrequency ablation(RFA) and investigate its techniques and skills, means and steps, and safety and efficacy. Methods Contrast-enhanced ultrasound-guided percutaneous RFA of thyroid adenomas were conducted on 202 patients by using an auto-controlled bi-polar electrode system. The indications of thyroid RFA,the optimal puncture route,the ways of anesthesia administration, protection of vital neck vessels and recurrent laryngeal nerve(RLN) and reduction of bleeding from core biopsy, indicators of ending ablation procedure following a complete ablation were investigated and analyzed. Resalts An adenoma smaller than 20 mm in maximal diameter was the optimal candidate for RFA. Either of two puncture routes could be selected upon the target lesion's location. Areas surrounding to the thyroid capsule needed adequate local anesthesia to kill pain. Liquid-isolating maneuver could effectively protect carotid artery and RLN from core needle cutting and electrode heating injury. Advanced block of supplying arteries with heating markedly reduced bleeding involved in the biopsy. Multipoint and multicenter ablation was essential to a complete coagulation. Filling-defect in the ablated adenoma on CEUS was the key sign to terminate ablation procedure. Conclusions Percutaneous bi-polar RFA was proved feasible, effective, safe and supermicroinvasive for treating thyroid adenoma under the way stated here of puncture and technical points and use of CEUS for monitoring.