Ultrafast Microfluidic PCR Thermocycler for Nucleic Acid Amplification.

The polymerase chain reaction (PCR) is essential in nucleic acid amplification tests and is widely used in many applications such as infectious disease detection, tumor screening, and food safety testing; however, most PCR devices have inefficient heating and cooling ramp rates for the solution, which significantly limit their application in special scenarios such as hospital emergencies, airports, and customs. Here, we propose a temperature control strategy to significantly increase the ramp rates for the solution temperature by switching microfluidic chips between multiple temperature zones and excessively increasing the temperature difference between temperature zones and the solution; accordingly, we have designed an ultrafast thermocycler. The results showed that the ramp rates of the solution temperature are a linear function of temperature differences within a range, and a larger temperature difference would result in faster ramp rates. The maximum heating and cooling ramp rates of the 25 µL solution reached 24.12 °C/s and 25.28 °C/s, respectively, and the average ramp rate was 13.33 °C/s, 6-8 times higher than that of conventional commercial PCR devices. The thermocycler achieved 9 min (1 min pre-denaturation + 45 PCR cycles) ultrafast nucleic acid amplification, shortening the time by 92% compared to the conventional 120 min nucleic acid amplification, and has the potential to be used for rapid nucleic acid detection.

Effect of X-ray radiation on the pharmacokinetics of apatinib in vivo in rats.

Purpose: The "radiotherapy-pharmacokinetic" ("RT-PK") phenomenon refers to the fact that radiation can significantly alter the pharmacokinetic behavior of a drug. At present, it is not clear whether there is an "RT-PK" phenomenon that can affect apatinib during concurrent chemoradiotherapy. In this study, we used a rat irradiation model to study the effects of X-ray radiation on absorption, tissue distribution, and excretion of apatinib. Method: Healthy Sprague-Dawley (SD) rats were randomly divided into control and radiation groups. The radiation group was given an appropriate dose of abdominal X-ray radiation, while the control group was not given irradiation. After 24 h of recovery, both groups were given apatinib solution 45 mg/kg by gavage. A quantitative LC-MS/MS method was developed to determine the concentration of apatinib in the rats, so as to compare the differences between the control and radiation groups and thus investigate the modulating effect of radiation on the pharmacokinetics of apatinib in rats. Results: After abdominal X-ray irradiation, the area under the curve (AUC0-t) of apatinib in rat plasma decreased by 33.8% and 76.3% at 0.5 and 2 Gy, respectively. Clearance (CL) and volume of distribution (Vd) increased and were positively correlated with radiation dose. X-ray radiation significantly reduced the concentration of apatinib in the liver and small intestine, and there was no tissue accumulation. In excretion studies, we found that X-ray radiation reduced the cumulative excretion of apatinib in feces and urine by 11.24% and 86.17%, respectively. Conclusion: Abdominal X-ray radiation decreased plasma exposure, tissue distribution, and excretion of apatinib in rats, suggesting that the RT-PK phenomenon affects apatinib. We speculate that this RT-PK phenomenon is closely related to changes in metabolic enzymes in vivo. In clinical practice, when apatinib is combined with radiotherapy, attention should be paid to adjusting the dose of apatinib and optimizing the treatment plan to alleviate the adverse effects of this RT-PK phenomenon.

Centromere-Specific Retrotransposons and Very-Long-Chain Fatty Acid Biosynthesis in the Genome of Yellowhorn (Xanthoceras sorbifolium, Sapindaceae), an Oil-Producing Tree With Significant Drought Resistance.

In-depth genome characterization is still lacking for most of biofuel crops, especially for centromeres, which play a fundamental role during nuclear division and in the maintenance of genome stability. This study applied long-read sequencing technologies to assemble a highly contiguous genome for yellowhorn (Xanthoceras sorbifolium), an oil-producing tree, and conducted extensive comparative analyses to understand centromere structure and evolution, and fatty acid biosynthesis. We produced a reference-level genome of yellowhorn, ∼470 Mb in length with ∼95% of contigs anchored onto 15 chromosomes. Genome annotation identified 22,049 protein-coding genes and 65.7% of the genome sequence as repetitive elements. Long terminal repeat retrotransposons (LTR-RTs) account for ∼30% of the yellowhorn genome, which is maintained by a moderate birth rate and a low removal rate. We identified the centromeric regions on each chromosome and found enrichment of centromere-specific retrotransposons of LINE1 and Gypsy in these regions, which have evolved recently (∼0.7 MYA). We compared the genomes of three cultivars and found frequent inversions. We analyzed the transcriptomes from different tissues and identified the candidate genes involved in very-long-chain fatty acid biosynthesis and their expression profiles. Collinear block analysis showed that yellowhorn shared the gamma (γ) hexaploidy event with Vitis vinifera but did not undergo any further whole-genome duplication. This study provides excellent genomic resources for understanding centromere structure and evolution and for functional studies in this important oil-producing plant.

Advantages of retrograde flexible ureteroscopy in determining the etiology of painless hematuria originating from the upper urinary tract.

The present study investigated the use of retrograde flexible ureteroscopy (RFU) in the discrimination of the etiology of hematuria that originates from the upper urinary tract (UUT). The present study collected retrospective data for patients who presented with hematuria and cystoscopy-detected bleeding from the UUT between June 2006 and August 2018 in Ningbo First Hospital. All patients accepted RFU to determine the etiology of hematuria. Data regarding imaging examinations, surgery, pathology and complications were also collected and analyzed. In total, 65 patients (males, 38; females, 27) with a mean age of 63 years underwent RFU to determine the etiology of hematuria originating from the UUT. Using RFU, UUT tumors were found in 29 cases. Stones, polyps and atypical hyperplasia were found in two cases, and a definite diagnosis was not found in three cases. There were 17 cases without obvious abnormalities and nine cases were unable to undergo RFU due to ureteral stenosis. In patients who could not be diagnosed by imaging examination, 34.4% (11/32) were diagnosed with urothelial carcinoma by RFU, and these results were also confirmed by postoperative pathology. In the present study, no patient had severe complications after RFU. The present results suggested RFU may be used as a sensitive method to diagnose UUT tumors (78.4%; 29/37) and has strong specificity. RFU could be performed as a routine examination for patients with hematuria from the UUT.

A matched-pair comparison of single plus one port versus standard extraperitoneal laparoscopic radical prostatectomy by a single urologist.

We conducted this study to report on our initial experience and assess the safety, feasibility, and efficacy of extraperitoneal single plus one port laparoscopic radical prostatectomy (SPOPL-RP), and determine whether it shows any objective advantage over standard laparoscopic radical prostatectomy. From June 2009 to September 2011, 15 extraperitoneal SPOPL-RPs were performed through a 2-3-cm subumbilical longitudinal incision and another 5-mm trocar placed at the McBurney point. This cohort was compared with 37 contemporary patients who underwent standard extraperitoneal laparoscopic radical prostatectomy performed by the same urologist. Peri- and postoperative outcomes, including continence, potency, and scar length, were statistically analyzed. The two groups were comparable with respect to patient demographics, estimated blood loss, drainage time, duration of catheterization, catheterization rate >14 days, complication rate, postoperative hospitalization, and postoperative functional and oncologic outcomes (p > 0.05). The SPOPL-RP procedures had a longer mean operative time (170.1 minutes vs. 139.5 minutes, p = 0.005), but with fewer patients requiring analgesics (20% vs. 54.1%, p = 0.038) and earlier resumption of oral intake (20.7 hours vs. 26.8 hours, p = 0.037). The mean scar length in the SPOPL-RP group was much smaller (3.4 cm vs. 5.8 cm, p = 0.000) owing to the significant reduction of the skin incision. The peri- and postoperative outcomes of SPOPL-RP for low-risk prostate cancer are comparable to those with the standard laparoscopic approach. In addition, SPOPL-RP provides better postoperative pain control, faster recovery of bowel function, and smaller scar length than standard laparoscopy, albeit with a longer operative time.

Robot-Assisted Radical Prostatectomy vs. Open Retropubic Radical Prostatectomy for Prostate Cancer: A Systematic Review and Meta-analysis.

Open retropubic radical prostatectomy (ORP) remains the "gold standard" for surgical treatment of clinically localized prostate cancer (PCa). Robot-assisted radical prostatectomy (RARP) is a robotic surgery used worldwide. The aim of this study is to collect the data available in the literature on RARP and ORP, and further evaluate the overall safety and efficacy of RARP vs. ORP for the treatment of clinically localized PCa. A literature search was performed using electronic databases between January 2009 and October 2013. Clinical data such as operation duration, transfusion rate, positive surgical margins (PSM), nerve sparing, 3- and 12-month urinary continence, and potency were pooled to carry out meta-analysis. Six studies were enrolled for this meta-analysis. The operation duration of RARP group was longer than that of ORP group (weighted mean difference = 64.84). There was no statistically significant difference in the transfusion rate, PSM rate, and between RARP and ORP (transfusion rate, OR = 0.30; PSM rate, OR = 0.94). No significant difference was seen in 3- and 12-month urinary continence recovery (3 months, OR = 1.32; 12 months, OR = 1.30). There was a statistically significant difference in potency between the 3- and 12-month groups (3 months, OR = 2.80; 12 months, OR = 1.70). RARP is a safe and feasible surgical technique for the treatment of clinically localized PCa owing to the advantages of fewer perioperative complications and quicker patency recovery.

Holmium laser versus conventional transurethral resection of the bladder tumor.

BACKGROUND: Transurethral resection of the bladder tumor (TURBT) remains the gold standard for non-muscle-invasive bladder cancer (NMIBC). Laser techniques have been widely used in urology. This analysis aimed to assess the safety and efficacy of holmium resection of the bladder tumor (HoLRBT) vs. TURBT. METHODS: A systemic search of MEDLINE, Embase, Web of Science, and The Cochrane Library as well as manual bibliography searches were performed to identify the relevant studies. The pooled estimates of operation time, obturator nerve reflex rate, bladder perforation rate, bladder irrigation rate, catheterization time, hospital stay, and one- and two-year recurrence free survivals were calculated. RESULTS: Five studies were enrolled into our meta-analysis. No significant difference was observed in the operation time between groups (weighted mean difference (WMD) 1.01, 95% confidential interval (95% CI) -3.52 - 5.54, P = 0.66). The significant difference in the obturator nerve reflex (OR 0.05, 95% CI 0.01 - 0.04, P = 0.004), bladder perforation (OR 0.14, 95% CI 0.03 - 0.61, P = 0.009), bladder irrigation (OR 0.13, 95% CI 0.04 - 0.45, P = 0.001), catheterization time (WMD -0.96, 95% CI -1.11 to -0.82, P < 0.00001), and hospital stay (WMD -1.46, 95% CI -1.65 to -1.27, P < 0.00001) showed advantages of HoLRBT over TURBT. The 2-year recurrence free survival rate favors the HoLRBT group (OR 1.46, 95% CI 1.02 - 2.11, P = 0.04). CONCLUSIONS: As a promising technique, HoLRBT is safe and efficient, and showed several advantages over TURBT. HoLRBT can be used as an alternative procedure for TURBT in terms of low-grade papillary urothelial carcinoma or low-grade early TNM-stage urothelial carcinoma.

Transperitoneal laparoscopic excision of primary seminal vesicle benign tumors: surgical techniques and follow-up outcomes.

OBJECTIVE: To assess the feasibility, safety, and efficacy of transperitoneal laparoscopic excision of primary seminal vesicle benign tumors (SVBTs) and summarize our experience with surgical techniques and follow-up outcomes of this rare condition. METHODS: This study included 6 patients who underwent transperitoneal laparoscopic excision of primary SVBTs between June 2005 and April 2011. A 5-port transperitoneal approach was used. The ipsilateral vas deferens was identified in the upper bulge of the retrovesical peritoneal reflection through a transverse approach and was dissected inwardly and used as a guide to the seminal vesicle tumor. Endoscopic Hem-o-lok clips (Teleflex Medical) were applied to control the vascular supply to the tumor base. With the contralateral vas deferens and seminal vesicle preserved, the tumor was removed together with the vas deferens and the adjoining ipsilateral seminal vesicle. The surgical procedures were successful in all 6 patients, without conversion to open surgery. The mean duration of surgery was 70 ± 16 minutes (range, 50-100 minutes), with unremarkable blood loss of less than 50 mL. The mean postoperative hospital length of stay was 5.2 ± 1.6 days (range, 4-8 days). No intra- or postoperative complications occurred. During a mean follow-up period of 42 ± 24 months (range, 12-82 months), all patients remained asymptomatic, with preserved function as reported by the patient, and there was no evidence of recurrence. CONCLUSION: Our study demonstrated that transperitoneal laparoscopic excision of primary SVBTs is a viable option for minimally invasive treatment of SVBT.

Evaluating the oncologic outcomes in 152 patients undergoing extraperitoneal laparoscopic radical prostatectomy.

BACKGROUND: Although many midterm oncologic data have been reported for extraperitoneal laparoscopic radical prostatectomy (ELRP) in western countries, few oncologic data of the extraperitoneal procedure was published in China. The aim of the study was to evaluate the oncologic outcomes of patients treated with ELRP in China. METHODS: From January 2005 to March 2010, a total of 152 consecutive patients diagnosed with clinically localized prostate cancer were included in this study and treated with ELRP. The patients were staged according to the TNM (tumor, nodes, metastases) system. Median and mean postoperative follow-up were 28.1 months and 27.0 months, respectively. The patients were retrospectively analyzed for progression-free survival. RESULTS: One hundred and twelve cases (73.7%) were postoperatively diagnosed as pT2 in, and 40 cases (26.3%) as pT3. Positive lymph nodes were shown in 5 patients (3.3%). Gleason score was < 7 in 49 men (32.2%), 7 in 69 men (45.4%), and > 7 in 34 men (22.4%). Positive surgical margins (PSM) were observed in 15 patients (9.9%), which included 32.0% of all pT3a cases and 46.7% of all pT3b cases, respectively. The overall prostate-specific antigen recurrence-free survival rate was 86% in all patients. The recurrence-free survival rates were 91.8% and 62.2% in pT2N0 patients and pT3N0 patients, respectively. Preoperative prostate-specific antigen, surgical margins, tumor stage, and lymph nodal status were identified as independent predictors of biochemical recurrence-free survival using multivariate Cox proportional hazard model. CONCLUSIONS: ELRP is a precise, safe and effective procedure at this particular Chinese institution. The prognostic power of prostate-specific antigen relapse after ELRP is not identical to that described previously with transperitoneal or open retropubic approaches.

Adrenocorticotropic hormone-producing pheochromocytoma: a case report and review of the literature.

Ectopic Cushing's syndrome caused by pheochromocytoma is rare. We reported a 15-year-old female patient who was admitted to hospital with typical Cushing's syndrome. She had not started menstruation. Her plasma adrenocorticotropic hormone (ACTH) and 24-hour urinary free cortisol levels were extremely high. Gonadal and progestational hormone levels were also abnormal. Abdominal computed tomography scans and enhanced scans revealed multiple irregular tumors in the right adrenal. Pelvic echogram showed an infantile uterus, while the ovaries were at an immature stage of development. Retroperitoneal laparoscopic right adrenalectomy was performed without intraoperative complications. Histology and immunohistochemistry of the tumor were consistent with pheochromocytoma. Retroperitoneal laparoscopic adrenalectomy is a safe procedure with satisfactory outcomes and allows for rapid recovery.

Transperitoneal laparoscopic excision of seminal vesicle cyst: a single-center experience.

BACKGROUND AND PURPOSE: Seminal vesicle cyst (SVC) is a rare disease and its treatment is still controversial. This article contains the largest series of transperitoneal laparoscopic excision of SVC to date, summarizing our surgical techniques and clinical experience with this disease. PATIENTS AND METHODS: From December 2003 to May 2010, seven patients received transperitoneal laparoscopic excision of SVC using a five-port transperitoneal approach. Nearly the total cyst was removed by only leaving a narrow strip of the cyst wall with the bilateral vas deferens and SV preserved completely. Pelvic CT or MRI was performed 3 and 6 months after surgery, and thereafter annually for at least 3 years. RESULTS: Transperitoneal laparoscopic excision of SVC was completed successfully in all seven patients without conversion to open surgery. The mean operative time was 73 minutes (range 60-100 min) with negligible blood loss (less than 20 mL). The mean postoperative hospital stay was 4.3 days (range 3-5 days). No intraoperative or postoperative complication occurred. The patients were followed up for a mean of 45 months (range 18-84 mos), during which they all remained symptom free with normal erectile and ejaculatory function without evidence of recurrence. CONCLUSION: Our study has demonstrated that transperitoneal laparoscopic excision of SVC is a safe, feasible, and efficacious procedure, and offers an excellent option for minimally invasive treatment of patients with SVC.

[Application of retroperitoneal laparoscopic partial nephrectomy in renal carcinoma patients with intermediate risk PADUA score].

OBJECTIVE: To study the application of retroperitoneal laparoscopic partial nephrectomy in renal carcinoma patients with intermediate risk PADUA score. METHODS: From April 2005 to June 2011, 79 cases (48 males and 31 females) of intermediate risk PADUA score (range from 8 to 9 score) renal cell carcinoma were retrospectively analyzed. Mean age was (54 ± 9) years, mean tumor size was (2.8 ± 0.8) cm in diameter, with 37 cases on the left side and 42 cases on the right side. Tumor located anteriorly in 35 cases, and 44 cases were located posteriorly. Preoperative imaging examinations showed tumor invasion of the collecting system was dislocated or infiltrated by tumor invasion were in 13 cases, renal sinus were involved in 5 cases, tumor located near the renal hilum were in 10 cases. All of the 79 patients received retroperitoneal laparoscopic partial nephrectomy. RESULTS: The 79 cases were operated successfully without conversion to open surgery, no severe perioperative complications. The mean operation time was (105 ± 24) minutes, and the median of operation time was 115 minutes (range from 80 - 180 minutes), and mean warm ischemia time (WIT) was (20 ± 5) minutes, and mean blood loss was (24 ± 8) ml; mean postoperative hospital stay was (5.2 ± 1.5) days. Postoperative urinary leakage in 3 cases, symptoms disappeared one week after indwelling catheterization and ureteral catheter. Serum creatinine transient increased in 7 cases after surgery, and fell to normal range within 6 weeks. In a mean follow up for (34 ± 12) months (range from 10 to 84 months), estimated glomerular filtration rate (eGFR) 6 months after operation was no statistical significance compared with preoperation in 77 cases, another 2 patients' eGFR decreased by 30% and 35%. Postoperative renal function remained in CKD3 period and CKD2 period were in 2 cases respectively, none of these cases were treated with hemodialysis, and the remaining patients with normal renal function after surgery, no tumor recurrence and metastasis during follow-up in all cases. CONCLUSIONS: Treatment of retroperitoneal laparoscopic partial nephrectomy in renal carcinoma patients with intermediate risk PADUA score is safe and effective, but its long-term effects still need to study with large samples compare and long-term follow-up.