Analysis of differential gene expression by SRAP-cDNA in mantle tissue of Meretrix petechialis with different external shell color.

Gene expression of two shell colors in Meretrix petechialis were analyzed by sequence related amplified polymorphism-cDNA to screen the associated molecular markers. The two shell color genomes of M. petechialis were amplified using combinations of 30 primers; 11 pairs of primers showed differential fragments, and by recovery, cloning and sequencing, 18 different differential sequences were obtained. The sequencing results were analyzed by BlastX. Only one fragment shared high homology with memory-related protein-2 and TonB-dependent receptor was found that related to shell color. Sequence characterized amplified region primers were designed according to the difference sequences, and PCR amplification was performed in both 'yellow' and 'red' M. petechialis. Four pairs of differential primers were obtained. Using the population to verify the four markers (Me1-Em2, Me2-Em3, Me4-The Em11 and Me4-Em12), it was found that Me1-Em2 and Me2-Em3 were positive in the 'yellow' and Me4-The Em11 and Me4-Em12 were positive in the 'red' M. petechialis populations. All four markers can, therefore, be used as M. petechialis shell color related markers. This provides a theoretical basis for studying shell color regulation in M. petechialis, which may help to reveal the underlying molecular mechanisms more comprehensively.

Pneumatic Lithotripsy versus Holmium Laser Lithotripsy in Percutaneous Nephrolithotomy for Patients with Guy's Stone Score Grade IV Kidney Stone.

BACKGROUND AND OBJECTIVE: The aim of this study was to compare the efficacy and reliability of holmium (Ho:YAG) laser lithotripsy (HLL) and pneumatic lithotripsy (PL) in percutaneous nephrolithotomy (PCNL) in the treatment of patients with Grade IV kidney stones based on Guy's Stone Score. STUDY DESIGN/MATERIALS AND METHODS: This retrospective study included 103 patients with Grade IV kidney stones out of 440 patients who underwent PCNL through HLL and PL in Second Affiliated Hospital of Shantou University Medical College, China, from January 2016 to December 2018. We analyzed preoperative, intraoperative, and postoperative variables of the patients to evaluate the efficacy and reliability of PCNL procedures. RESULTS: Patients were categorized as Grade I, II, III, and IV, and the patients of each grade were 85 (19.32%), 39 (8.86%), 213 (48.41%), and 103 (23.41%), respectively. In Grade IV, the total operative time (min) for the PL and HLL groups was 137.7 ± 47.79 and 134.27 ± 53.38, respectively (p = 0.744). The variation in laboratory examination values including ΔHGB (g/L), ΔHCT, ΔPCT, and ΔCr (µmol/L) for PL and HLL groups was 19 ± 11.23/12 ± 15.42 (p = 0.012), 0.057 ± 0.034/0.038 ± 0.045 (0.009), 0.027 ± 0.034/0.026 ± 0.034 (0.702), and 3.07 ± 17.4/20.54 ± 65.93 (0.692), respectively. The postoperative hospitalization day was 8.94 ± 4.2 and 7.73 ± 2.75 (p = 0.015), respectively. As for the stone-free rate (SFR), the SFRs for PL and HLL were 48.15% (n = 39/81) and 59.09% (n = 13/22) (p = 0.363), respectively. CONCLUSIONS: HLL showed a comparable advantage of not only decreased postoperative hemoglobin and hematocrit but also fewer postoperative hospitalization days. Based on the results of our retrospective study, for those Grade IV kidney stone patients who have a risk of bleeding before PCNL operation, HLL can be a considerable treatment option. Besides, in consideration of reducing human care cost, HLL which showed fewer hospitalization days, would be more welcome by patients.

Genetic diversity and differentiation of nine populations of the hard clam (Meretrix petechialis) assessed by EST-derived microsatellites

BACKGROUND: Meretrix petechialis is one of the commercially important marine bivalves. In this study, we selected six highly polymorphic EST-derived microsatellite markers to assess the genetic diversity and population differentiation on nine wild populations of Meretrix petechialis. RESULTS: The number of alleles detected per-locus ranged from 4 to 30 (mean NA = 27.5) with a total of 165 alleles. The mean value of observed and expected heterozygosities varied from 0.717 to 0.861 and from 0.797 to 0.856, respectively. Meanwhile, the result of Neighbor-joining and overall FST = 0.214 (P < 0.01) reveled that M. petechialis populations from GX are the farthest populations, a certain degree of genetic variation among individuals in each population and the genetic differentiation is significant. CONCLUSIONS: GX population has high genetic diversity among individual, and there are certain differences in genetic characteristics among different populations. This study will provide a basis for the domestication and cultivation of genetic diversity of M. petechialis population and the protection of clam germplasm resources.

Genetic diversity and population structure of Meretrix petechialis in China revealed by sequence-related amplified polymorphism markers.

Genetic variation in nine stocks of Meretrix petechialis collected from China was analyzed using sequence-related amplified polymorphism (SRAP) markers. Eight primer pairs produced 132 polymorphic loci with an average of 16.5 loci per primer pair. A population from Jiangsu had the highest percentage of polymorphic loci at 27.27%, suggesting that these resources had a rich genetic diversity. The Nei's gene diversity of the nine populations ranged from 0.0647 to 0.0793; a population from Shandong was the lowest and a population from North Korea the highest. The Shannon's information index was between 0.1023 and 0.1202, with the lowest in the Shandong population and the highest in the Jiangsu population. The Nei's unbiased genetic distance between the nine populations was 0.0243-0.0570 and the genetic similarity was 0.9446-0.9760; the genetic distance between Guangxi and Shandong populations was the furthest (0.0570) and the genetic distance between Shandong and Jiangsu populations was the closest (0.0243). Nei's gene diversity analysis indicated that the genetic variance was mainly found within individual geographical populations, and the analysis of molecular variance revealed low but significant genetic differentiation among local and regional populations. The limited gene flow (Nm = 0.555) was inferred as a major reason for the extent of genetic differentiation in M. petechialis. The results obtained here indicated that M. petechialis have high degree of genetic diversity and the potential of further breeding with excellent germplasm resources. This study provides a scientific basis for the protection of germplasm resources and the breeding of M. petechialis.

Genetic variation and population structure of different geographical populations of Meretrix petechialis based on mitochondrial gene COI.

The hard clam (Meretrix petechialis) is a commercially important shellfish in China. To provide valuable insights into management and conservation of M. petechialis, we investigated the genetic variation and population structure of M. petechialis by analysing samples from nine geographical populations. In this study, the genetic diversity and differentiation of nine populations of M. petechialis were assessed using the mitochondrial cytochrome oxidase subunit I (mtCOI) gene. A total of 90 COI sequences were obtained and each COI sequence was 699 bp in length. Fifty-one haplotypes were identified with 10 haplotypes shared among populations. The haplotype diversity was highest in Fujian, Panjin and Jiangsu (0.9778 ± 0.0540) and lowest in Dalian (0.7778 ± 0.1374). The nucleotide diversity was highest in Panjin (0.453401 ± 0.240463) and lowest in Jiangsu (0.006213 ± 0.004141). Neutral test (Fu's Fs) and mismatch distribution analysis revealed that the hard clam experienced a population expansion event. Analysis of molecular variance (AMOVA) indicated that 91.7% of the genetic variance was within populations and 0.52% of the variance was among populations, demonstrating significant genetic differentiation among populations (P < 0.05). The neighbour-joining tree showed that the haplotypes were not clustered according to the geographical location, but some haplotypes from the same or neighbouring locations grouped together. The results obtained in this study provide useful information on the genetic diversity and population structure of M. petechialis and shed light on the management and protection of resources of M. petechialis in the northwestern Pacific.

Vimentin Overexpressions Induced by Cell Hypoxia Promote Vasculogenic Mimicry by Renal Cell Carcinoma Cells.

Vasculogenic mimicry (VM), the novel approach for tumor cells to obtain blood supply, was reported to be involved in antiangiogenic resistance and poor prognosis in renal cell carcinoma (RCC). However, the molecular mechanisms underlying VM formed by RCC cells are still not clearly depicted. In the present study, we found that OS-RC-2 acquired the VM forming ability accompanied with the increased expressions of Vimentin and AXL and decreased expression of E-Cadherin by CoCl2 treatment. Downregulation of Vimentin by siRNA severely impaired the capability of OS-RC-2 and 786-O to form VM structures induced by cell hypoxia in vitro. Moreover, knockdown of Vimentin inhibited cell migration and invasion, which could be prompted by hypoxia induction in RCC cells. In our clear cell RCC tissues, we found that VM was positively correlated with Vimentin overexpression and both predicted poor prognosis. In conclusion, Vimentin plays an important role in hypoxia induced VM formation of RCC cells and targeted Vimentin might be beneficial for RCC therapy.

Retroperitoneoscopic nephrectomy for a horseshoe kidney with hydronephrosis and inflammation: A case report.

RATIONALE: A horseshoe kidney is the most common congenital fusion abnormality in the kidney, occurring in approximately 1 in 400 live births. Several complications including renal malignancies, ureteropelvic junction obstruction, urolithiasis, vesicoureteral reflux, and hydronephrosis can occur in this patient population. PATIENT CONCERNS: A 28-year-old woman was admitted to hospital because of chronic left low back pain. Microscopic hematuria was not seen. Computed tomography showed the horseshoe kidney and left hydronephrosis. DIAGNOSES: On the basis of these findings and clinical manifestations, the final diagnosis was the horseshoe kidney with left renal hydronephrosis and inflammation. INTERVENTIONS: A retroperitoneoscopic nephrectomy on the left kidney was performed. OUTCOMES: Histopathological examination of the specimen confirmed massive dilatation of the pelvicaliceal system and chronic pyelonephritic inflammation. The patient was discharged on the 7th postoperative day with no complications and no back pain. She remained well at 3 months with normal activity and good cosmetic result. LESSONS: Retroperitoneoscopic nephrectomy can be a safe and minimally invasive surgery for horseshoe kidney treatment.

MTHFD2 Overexpression Predicts Poor Prognosis in Renal Cell Carcinoma and is Associated with Cell Proliferation and Vimentin-Modulated Migration and Invasion.

BACKGROUND/AIMS: To investigate the role of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) in the clinical prognosis and cell biology of renal cell carcinoma (RCC). METHODS: A total of 137 RCC tissues were evaluated by immunohistochemistry. The relationship between MTHFD2 overexpression and clinical parameters and vimentin expression was assessed. Kaplan-Meier curves and the log-rank test were applied for survival analysis according to MTHFD2 and vimentin expression in RCC tissues. The expression of MTHFD2 mRNA and protein was examined by quantitative reverse transcription PCR and western blotting, respectively. To determine further the biological activity of MTHFD2 in RCC, 786-O cells were transfected with short hairpin RNA specifically targeting MTHFD2 (shMTHFD2) with or without tumor necrosis factor (TNF)-α stimulation. Cell proliferation, cell migration and invasion and drug sensitivity were subsequently assessed using Cell Counting Kit-8, wound healing, and Transwell assays. RESULTS: Immunohistochemical analysis demonstrated that both MTHFD2 and vimentin overexpression was positively associated with clinical staging, pathological grade, and poor overall survival (all P < 0.05). MTHFD2 expression was closely correlated with vimentin overexpression in RCC (r = 0.402, P < 0.001). After knocking down MTHFD2 expression in 786-O cells, decreased cell proliferation, migration, and invasion were observed and accompanied by the reduced expression of vimentin. The effects of MTHFD2 down-regulation could be partially restrained by TNF-α treatment. Vimentin expression and cell migration and invasion, but not cell proliferation, were reversed by TNF-α stimulation. Furthermore, treatment of 786-O cells with shMTHFD2 increased their sensitivity to chemotherapy drugs. CONCLUSION: The current results demonstrated that MTHFD2 was overexpressed in RCC and associated with poor clinical characteristics, vimentin expression, and cellular features connected to malignant disease, thus, implicating MTHFD2 as a potential target for RCC therapy.

Dual-centre randomized-controlled trial comparing transurethral endoscopic enucleation of the prostate using diode laser vs. bipolar plasmakinetic for the treatment of LUTS secondary of benign prostate obstruction: 1-year follow-up results.

PURPOSE: Bipolar endoscopic enucleation of the prostate (BEEP) was recommended by the 2016 EAU guidelines as the first choice of surgical treatment in men with a substantially enlarged prostate and moderate-to-severe lower urinary tract symptoms. The main aim of this study was to compare a modified diode laser enucleation of the prostate (DiLEP) to BEEP. METHODS: A total of 114 patients with prostate (20-160 mL) were randomized 1:1 into either DiLEP or BEEP in a dual-centre, non-inferiority-design randomized-controlled trial. The primary outcomes included Qmax and IPSS at 12 months. Non-inferiority was evaluated by comparing the two-sided 95% CI for the mean differences of Qmax and IPSS. Secondary endpoints included other perioperative parameters, postoperative micturition variables, and complication rate. RESULTS: A total of 111 patients (97%) had completed the intent-to-treat analysis, The results showed that DiLEP was comparable to BEEP regarding Qmax (28.0 ± 7.0 vs. 28.1 ± 7.2 mL/s) and IPSS (3.0 ± 2.2 vs. 2.9 ± 2.6) at 12 months, the non-inferiority was met for both Qmax and IPSS. There were also no significant difference between two groups regarding tissue removal rate (71.8 vs. 73.8%), hemoglobin decrease (0.33 ± 0.66 vs. 0.36 ± 0.75 g/dL), sodium decrease (1.0 ± 2.7 vs. 0.3 ± 2.9 mmol/L), and Clavien III complications (5.3 vs. 1.8%) at 12 months. CONCLUSIONS: This DiLEP is an anatomical endoscopic enucleation technique for the treatment of benign prostatic hyperplasia, it is non-inferior to BEEP regarding Qmax and IPSS at 12 months postoperatively.

Expression pattern of asialoglycoprotein receptor in human testis.

During acute or chronic hepatitis B virus (HBV) infection, the virus can invade the male reproductive system, pass through the blood-testis barrier and integrate into the germ line, resulting in abnormal spermatozoa. However, the pathway remains unclear. The asialoglycoprotein receptor (ASGR), a potential receptor for HBV, is mainly distributed in hepatocytes. We have examined the distribution of ASGR in human testis and found it in the seminiferous tubules and interstitial region but its enrichment in human testis is much lower than that in liver. By multiple immunoenzyme histochemistry staining, ASGR was precisely co-localized with vimentin (Sertoli cell marker) but not proliferating cell nuclear antigen (spermatogonial cell marker) in testis tissue. ASGR was expressed in human Leydig cells, stromal cells in the seminiferous tubules and Sertoli cells but seldom in spermatogonial cells. Therefore, ASGR could provide HBV with access to the luminal compartment of human testis. The mechanism by which HBV invades germ cells remains unknown.

[Gossypol decreases the expression of connexin 43 in Sertoli cells].

OBJECTIVE: To study the effect of gossypol on the expression of connexin 43 (CX43) in Sertoli cells. METHODS: TM4 Sertoli cells were cultured and treated with gossypol at the concentrations of 1.25, 2.5, 5 and 10 micromol/L for 6, 12, 24 and 48 hours. The cytotoxicity of gossypol was assessed by CCK-8 assay, and the expression of CX43 in the normal TM4 Sertoli cells and in those treated with different concentrations of gossypol for different times was detected by RT-PCR and immunofluorescence analysis. RESULTS: Semiquantitative RT-PCR and immunofluorescence analysis showed the expression of CX43 in the normal TM4 cells. At 24 hours of exposure to gossypol, the expression began to decrease gradually with the prolonging of time and the increasing concentration of gossypol (P < 0.05). CONCLUSION: Gossypol reduces the expression of CX43 in TM4 Sertoli cells, which might underlie the mechanism of its antifertility action.

[Simultaneous determination of 128 pesticide residues in milk by liquid chromatography-tandem electrospray mass spectrometry].

A method has been developed for the simultaneous determination of 128 pesticide residues in milk by liquid chromatography-tandem electrospray mass spectrometry (LC-ESI-MS/MS). A total of 10 mL milk was extracted with 20 mL acetonitrile (plus 4 g magnesium sulfate and 1 g NaCl) for two times. The concentrated supernatant was cleaned-up by a C18 solid phase extraction cartridge to remove the lipophilic compounds. The eluate was concentrated to about 0.5 mL in 45 degrees C water bath with nitrogen. The 1 mL mixture of acetonitrile and water (3:2, v/v) was added into the sample and mixed thoroughly for 30 s. The sample solution was filtered by 0.2 microm millipore filters before LC-ESI-MS/MS determination. This method was evaluated at the two spiked levels of 2 times and 8 times of limits of the detection (LODs) in 5 parallel experiments, respectively. The results showed that the average spiked recoveries of 128 pesticides at the low spiked level (0.14 microg/L-0.62 mg/L) were from 60.4% to 118.4% with relative standard deviations (RSDs) of 2.1%-24.3%, and the average spiked recoveries at the high spiked level (0.56 microg/L-2.48 mg/L) were from 64.4% to 118.5% with the RSDs of 1.3%-24.1%. The method showed a good linear relationship with the correlation coefficients over 0.99. The LODs were from 0.07 microg/L to 0.31 mg/L. This method is selective, accurate and easy to operate.