Application of robotic surgery in urology / 中华内分泌外科杂志

Robot assisted laparoscopic surgery is a more advanced minimally invasive procedure with distinct advantages over conventional laparoscopic surgery. Since the introduction of Da Vinci robotic equipment in 2006, a large number of robotic surgeries have been performed in China, especially in the field of Urology, and robotic surgery has been widely used in the treatment of adrenal tumor, renal tumor, bladder cancer, prostate cancer, and other diseases. Based on rich experience of more than 3000 cases of robotic surgery in our center, we summarize the status quo of urologic robotic surgery and discuss its development prospect.

Current progress of computational modeling for guiding clinical atrial fibrillation ablation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Atrial fibrillation (AF) is one of the most common arrhythmias, associated with high morbidity, mortality, and healthcare costs, and it places a significant burden on both individuals and society. Anti-arrhythmic drugs are the most commonly used strategy for treating AF. However, drug therapy faces challenges because of its limited efficacy and potential side effects. Catheter ablation is widely used as an alternative treatment for AF. Nevertheless, because the mechanism of AF is not fully understood, the recurrence rate after ablation remains high. In addition, the outcomes of ablation can vary significantly between medical institutions and patients, especially for persistent AF. Therefore, the issue of which ablation strategy is optimal is still far from settled. Computational modeling has the advantages of repeatable operation, low cost, freedom from risk, and complete control, and is a useful tool for not only predicting the results of different ablation strategies on the same model but also finding optimal personalized ablation targets for clinical reference and even guidance. This review summarizes three-dimensional computational modeling simulations of catheter ablation for AF, from the early-stage attempts such as Maze III or circumferential pulmonary vein isolation to the latest advances based on personalized substrate-guided ablation. Finally, we summarize current developments and challenges and provide our perspectives and suggestions for future directions.

Inhibition of Rac1-dependent forgetting alleviates memory deficits in animal models of Alzheimer's disease

Accelerated forgetting has been identified as a feature of Alzheimer's disease (AD), but the therapeutic efficacy of the manipulation of biological mechanisms of forgetting has not been assessed in AD animal models. Ras-related C3 botulinum toxin substrate 1 (Rac1), a small GTPase, has been shown to regulate active forgetting in Drosophila and mice. Here, we showed that Rac1 activity is aberrantly elevated in the hippocampal tissues of AD patients and AD animal models. Moreover, amyloid-beta 42 could induce Rac1 activation in cultured cells. The elevation of Rac1 activity not only accelerated 6-hour spatial memory decay in 3-month-old APP/PS1 mice, but also significantly contributed to severe memory loss in aged APP/PS1 mice. A similar age-dependent Rac1 activity-based memory loss was also observed in an AD fly model. Moreover, inhibition of Rac1 activity could ameliorate cognitive defects and synaptic plasticity in AD animal models. Finally, two novel compounds, identified through behavioral screening of a randomly selected pool of brain permeable small molecules for their positive effect in rescuing memory loss in both fly and mouse models, were found to be capable of inhibiting Rac1 activity. Thus, multiple lines of evidence corroborate in supporting the idea that inhibition of Rac1 activity is effective for treating AD-related memory loss.

The role of cell-crystal reaction mediated inflammation in the formation of intrarenal calcium oxalate crystals / 中华外科杂志

Calcium oxalate nephrolithiasis is the common disease of urinary surgery, its exact pathogenesis is still unclear.It is believed that the renal inflammatory injury induced by cell-crystal reaction plays an important role in the formation of intrarenal calcium oxalate crystals. Recent studies indicated that inflammation induced by cell-crystal reaction can cause renal cell damage, stimulate intracellular expression of NADPH oxidase, trigger the massive production of reactive oxygen species, activate nuclear factor-κB signaling pathway, release a large number of inflammatory factors, and cause inflammatory cascade effect of the kidney, thus promoting the accumulation, nucleation and growth of calcium salt crystals, eventually leading to the formation of intrarenal crystals and even stones. In this process, the regulatory factors and mechanisms involved include macrophages, NLRP3-high mobility group box-1 protein inflammation network, fetuin A, autophagy activation and other factors.

Isolation and identification of exosomes secreted from HK-2 cells exposed to high oxalate / 中华泌尿外科杂志

Objective To investigate the method for isolating and identification exosomes from the HK-2 cells exposed to high oxalate.Methods HK-2 cells were cultured to serum-free culture medium and treated with oxalate at the concentration from 0 mmol/L to 10.00 mmol/L for 48 h.CCK-8 assays were performed to measure the cells proliferation.Combined the cell morphology,observed under inverted microscope with statistical analysis,we finally 2.00 mmol/L oxalic acid as the experimental concentration.The HK-2 cell was exposed to 2.00 mmol/L oxalate for 48 h.The supernatants were collected.Exosomes were isolated and purified from the supernatants by ultracentrifugation.Transmission electron microscopy (TEM) was used to observe the morphology of isolated exosomes.Particle size of exosomes were detected with Nanosight technology.Western blot analysis was used to examine the experession of HSP70,CD63.Results CCK-8 assay showed that the cell viability in each group,including (100.0 ± 4.0) ％ in 0 mmol/L group;(97.7 ± 1.5)％ in the 0.25 mmol/L group;(97.3 ±2.1)％ in the 0.50 mmol/L group;(87.7 ± 2.1) ％ in the 1.00 mmol/L group;(76.0 ± 1.0) ％ in the 2.00 mmol/L group;(58.1 ± 2.6) ％ in the 4.00 mmol/L group;(52.7 ± 1.5) ％ in the 5.00 mmol/L group;(37.7 ± 3.2) ％ in the 8.00 mmol/L group;(31.3 ±2.0)％ in the 10.00 mmol/L group.The isolated exosomes demonstrated round or oval shape under TEM.The peak particle size was 56 nm,which the overall mean particle size was 87 nm.Those and particles with a diameter between 30-150 nm accounted for 91.2％.In this experiment,The expression of HSP70,CD63 could be detected in the isolated exosomes.However,only the expression of HSP70 could be detected in the HK-2 cells.Conclusions Under the treatment of 2 mmol/l oxalate for 48 hours,Ultracentrifugation can be used to isolate and purify exosomes efficiently from the HK-2 cells.This is helpful for further study of exosome as mediator of cell-to-cell communication.

Isolation and identification of exosomes secreted from HK-2 cells exposed to high oxalate / 中华泌尿外科杂志

Objective To investigate the method for isolating and identification exosomes from the HK-2 cells exposed to high oxalate.Methods HK-2 cells were cultured to serum-free culture medium and treated with oxalate at the concentration from 0 mmol/L to 10.00 mmol/L for 48 h.CCK-8 assays were performed to measure the cells proliferation.Combined the cell morphology,observed under inverted microscope with statistical analysis,we finally 2.00 mmol/L oxalic acid as the experimental concentration.The HK-2 cell was exposed to 2.00 mmol/L oxalate for 48 h.The supernatants were collected.Exosomes were isolated and purified from the supernatants by ultracentrifugation.Transmission electron microscopy (TEM) was used to observe the morphology of isolated exosomes.Particle size of exosomes were detected with Nanosight technology.Western blot analysis was used to examine the experession of HSP70,CD63.Results CCK-8 assay showed that the cell viability in each group,including (100.0 ± 4.0) ％ in 0 mmol/L group;(97.7 ± 1.5)％ in the 0.25 mmol/L group;(97.3 ±2.1)％ in the 0.50 mmol/L group;(87.7 ± 2.1) ％ in the 1.00 mmol/L group;(76.0 ± 1.0) ％ in the 2.00 mmol/L group;(58.1 ± 2.6) ％ in the 4.00 mmol/L group;(52.7 ± 1.5) ％ in the 5.00 mmol/L group;(37.7 ± 3.2) ％ in the 8.00 mmol/L group;(31.3 ±2.0)％ in the 10.00 mmol/L group.The isolated exosomes demonstrated round or oval shape under TEM.The peak particle size was 56 nm,which the overall mean particle size was 87 nm.Those and particles with a diameter between 30-150 nm accounted for 91.2％.In this experiment,The expression of HSP70,CD63 could be detected in the isolated exosomes.However,only the expression of HSP70 could be detected in the HK-2 cells.Conclusions Under the treatment of 2 mmol/l oxalate for 48 hours,Ultracentrifugation can be used to isolate and purify exosomes efficiently from the HK-2 cells.This is helpful for further study of exosome as mediator of cell-to-cell communication.

The Progress of Non-invasive Screening Methods for Colorectal Cancer / 现代生物医学进展

Early detection and treatment of high-risk adenomas and colorectal cancer (CRC) can reduce mortality of this disease.CRC screening is aimed at minimizing its harm and colonoscopy is presently the gold standard for it.However,colonoscopy needs bowel preparation and is invasive with high risk of intestinal perforation,causing a bad compliance,which is unfavorable to its popularization and application.Recently,non-invasive detection methods for CRC have gone through a rapid development.Tests based on CRC-related biomarkers in fecal and blood samples provide new options for non-invasive CRC screening.However,detection methods for these biomarkers still need further research and improvement because of the complex composition of feces and blood.In the two aspects of fecal tests and blood tests,the progress of recent studies on non-invasive screening methods for CRC was reviewed in this article.

Ultra-sensitive quantification of the colorectal cancer-specific NDRG4 gene methylation levels in stool / 医学研究生学报

Objective The NDRG4 gene methylation in stool is a candidate biomarker for non?invasive diagnosis of colorectal cancer. However, the traditional methods for methylation detection could not be well applied to stool samples due to the low sensitivity and low specificity. The aim of this study was to develop a highly sensitive and specific method for quantifying the methylated NDRG4 gene in stools. Methods Forty one stool samples were collected from 12 colorectal cancer patients, 4 adenoma patients and 25 nor?mal persons. The invasive reaction was combined with real?time PCR and the relative quantification was performed by 2-ΔCT method to develop the highly sensitive and specific methylated DNA detection method, which was used for detecting NDRG4 methylation levels in 41 of stool samples. Results The sensitivity of the method was as low as 10 copies of methylated NDRG4 gene fragments. The specificity was high enough to distinguish 0.01% of methylated fragments from un?methylated fragments and 105 copies of unmethylated NDRG4 fragments gave noamplification signals. The detection results from 41 of stool samples showed that detection rate of the NDRG4 gene in stool from adenoma and colorectal cancer groups had a significant difference comparing to that from the normal group. Conclusion The 2-ΔCT method could accurately quantify the methylation levels of the NDRG4 gene in stool samples, and provide an efficient tool for non?invasive colorectal cancer detection.

Detection of Single Nucleotide Polymorphism by Real-time Polymerase Chain Reaction Coupled with High Specific Invader Assay in Single Tube / 分析化学

A method for the real-time polymerase chain reaction ( PCR ) coupled with high specific invader assay to detect single nucleotide polymorphism ( SNP) was established. To reduce the background signal, the amount of flap endonuclease 1 ( FEN1 enzyme ) and wild-type detection probe was optimized. Under the optimum conditions including 0. 05 μmo/L invasive oligonucleotide probe, 0. 125 μmol/L wild-type detection probe, 0. 5 μmol/L mutation detection probe, 0. 25 μmol/L each fluorescence resonance energy transfer (FRET) probe and 1. 5 U FEN1, the background signal of wild-type sample and mutation sample was dramatically decreased and the background interference to the detecting results was thus eliminated. A total of 21 cases of aldehyde dehydrogenase-2*2 ( ALDH2*2 ) , 19 cases of cytochrome p450 2 C19*2 ( CYP2 C19*2 ) and 19 cases of CYP2C19*3 were analyzed with the established method, and the genotypes of ALDH2*2 were 10 cases of GG homozygote, 8 cases of GA heterozygote and 3 cases of AA homozygote; the genotypes of CYP2C19*2 were 9 cases of GG homozygote, 8 cases of GA heterozygote and 2 cases of AA homozygote;and the genotypes of CYP2C19*3 were 18 cases of GG homozygote and 1 case of GA heterozygote. These results were consistent with those by pyrosequencing. The established method was specific, simple, short time-consuming and low cost, and could be used for the detection of SNP genotyping with non-polluting in single closed tube.

Induction of osteogenic differentiation of human renal fibroblasts in vitro / 中国病理生理杂志

AIM:To investigate the effects of osteogenic induction media and the medias containing different concentration of calcium on the induction of osteogenic differentiation of human renal fibroblasts in vitro.METHODS: Culturedhuman renal fibroblasts were divided into 5 groups in this experiment: osteogenic induction group (osteogenic inductionmedia), CaⅠgroup (0.5 mmol/L Ca2 + media), CaⅡgroup (1.5 mmol/L Ca2 + media), Ca Ⅲ group (2.5 mmol/LCa2 + media) and control group (PBS).The cell activity in each groups was measured by MTT assay .At 9th day, the cellcalcium Alizarin red S staining and alkaline phosphatase (ALP) Gomori calcium cobalt staining were performed respectivelyto observe the formation of calcium nidus and the expression of ALP .In addition, the expression of Runt-related transcriptionfactor 2 (Runx2) at mRNA and protein levels was determined by real -time PCR and Western blot, respectively.RE- SULTS: The remarkable positive signs which represented the formation of calcium nidus and the deposit of calcium objectsin all experiment groups were observed .The mRNA and protein expression of Runx2 in osteogenic induction group increasedin accordance with the induction time .Compared with control group, the mRNA and protein expression of Runx2 inthe CaⅠ ~Ⅲ groups increased gradually in a calcium concentration dependent manner at the 9th induction day.CON- CLUSION: Human renal interstitial fibroblasts show the potential activity in osteogenic differentiation induced by osteogen -ic induction media or high level calcium in vitro, which may be account for the cytological formation of the Randall ’splaque in the kidney.

Genotyping of Alcohol Dehydrogenase Gene by Pyrosequencing Coupled with Improved Linear_after_the_Exponential Polymerase Chain Reaction Using Human Whole Blood as Starting Material / 分析化学

Pyrosequencing is one of the important genetic polymorphism detection methods currently, but the complicated pretreatment procedure limits its application in clinical test. To simplify the whole process of pyrosequencing, on the basis of the linear_after_the_exponential_polymerase chain reaction ( LATE_PCR) , we improved the primer design method of LATE_PCR, increased the length and the concentration of the excess primer, applied direct amplification technology with whole blood, and established a whole blood_imLATE_PCR method based on common rTaq polymerase and “HpH Buffer” ( High pH buffer ) . The amplification system was optimized, and the influences of blood anticoagulant and the amount of whole blood template were investigated. The single stranded template for the pyrosequencing was obtained by PCR amplification using a single tube in one_step process, and the alcohol dehydrogenase gene polymorphisms of 24 clinical blood samples were then detected successfully. The results could be used to guide clinical individualized medication. The genotypes of ADH1B locus of 24 samples were 6 cases of AA homozygote, 14 cases of AG heterozygote, and 4 cases of GG homozygote. The genotypes of ADH1C were 20 cases of GG homozygote, 4 cases of AG heterozygote, and no cases of AA homozygote.

Effect of intracavitary therapy on acute pulmonary embolism / 中国综合临床

Objective To explore the effect of intracavitary therapy on acute pulmonary embohsm.Methods Fifteen patients were selected as our subjects,who suffered acute pulmonary embolism and received percutaneous catheter thrombus crashing and catheter directed thrombolysis in Beijing Military.Region General Hospital from January 2009 to June 2011.Local injection of Urokinase was performed with a total amount of 500 000 U in catheter directed thrombolysis.After thrombolysis,low molecular Heparin was administered to patients for 7-10 days and oral administration of Warfarin was performed for 3-6 months.Clinical symptoms,improvement of physical signs,complications,changes of mean pulmonary arterial pressure(mPAP) and arterial partial pressure of oxygen(PO2),and the opening of pulmonary artery were recorded.Results The pulmonary arteries of the 12 patients were completely opened,and partially opened in 3 patients.The effective rate was 100％ (15/15).mPAP was reduced from (40.07 ±5.97) mmHg to (20.00 ±4.66) mmHg (t =-1.128,P ＜ 0.05),PO2 was increased from (50.26 ± 9.30) mmHg to (80.49 ± 9.04) mmHg (t =1.246,P ＜ 0.05).Patients were followed-up for 3-6 months and no recurrence case was seen.Conclusion The interventional therapy is effective,safe and practicable in the treatment of acute pulmonary embolism.

Preparation and Identification of Cetuximab-β-Glucosidase Conjugates / 天津医药

Objective To prepare cetuximab-β-glucosidase conjugates and to identify its enzymatic activity and an?tibody activity. Methods Cetuximab andβ-glucosidase were crosslinked by Sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC). Cetuximab-β-glucosidase conjugates and its enzymatic and antibody activity were examined by non-reduced SDS-PAGE, colorimetry and indirect immunofluorescence assay. Results We can see clear bands ofβ-glucosidase, cetuximab, cetuximab-β-glucosidase conjugates through electropherogram. Although the en?zymatic activity of cetuximab-β-glucosidase conjugates was lower than that ofβ-glucosidase (U/L:672.97±46.19 vs 869.50± 57.28,t=5.972,P<0.05) shown by colorimetry assay, it still maintain good enzymatic activity. Under fluorescence micro?scope, we can see the conjugates interacted with human bladder cancer EJ cells are in a red fluorescence. Conclusion Ce?tuximab,β-glucosidase were crosslinked successfully by Sulfo-SMCC without altered its enzymatic and antibody activity.

Model-based comparative prediction of transcription-factor binding motifs in anabolic responses in bone / 基因组蛋白质组与生物信息学报·英文版

Understanding the regulatory mechanism that controls the alteration of global gene expression patterns continues to be a challenging task in computational biology. We previously developed an ant algorithm, a biologically-inspired computational technique for microarray data, and predicted putative transcription-factor binding motifs (TFBMs) through mimicking interactive behaviors of natural ants. Here we extended the algorithm into a set of web-based software, Ant Modeler, and applied it to investigate the transcriptional mechanism underlying bone formation. Mechanical loading and administration of bone morphogenic proteins (BMPs) are two known treatments to strengthen bone. We addressed a question: Is there any TFBM that stimulates both "anabolic responses of mechanical loading" and "BMP-mediated osteogenic signaling"? Although there is no significant overlap among genes in the two responses, a comparative model-based analysis suggests that the two independent osteogenic processes employ common TFBMs, such as a stress responsive element and a motif for peroxisome proliferator-activated receptor (PPAR). The post-modeling in vitro analysis using mouse osteoblast cells supported involvements of the predicted TFBMs such as PPAR, Ikaros 3, and LMO2 in response to mechanical loading. Taken together, the results would be useful to derive a set of testable hypotheses and examine the role of specific regulators in complex transcriptional control of bone formation.