The effect and safety assessment of monitoring ethanol concentration in exhaled breath combined with intelligent control of renal pelvic pressure on the absorption of perfusion fluid during flexible ureteroscopic lithotripsy.

PURPOSE: Previously, we designed a ureteral access sheath with the capability of renal pelvic pressure (RPP) measurement and a medical perfusion and aspiration platform, allowing for the intelligent control of RPP. However, the effect of different RPP levels on perfusion fluid absorption remains unclear. This randomized controlled trial aimed to investigate the effects of exhaled ethanol concentration monitoring and intelligent pressure control on perfusion fluid absorption during flexible ureteroscopic lithotripsy. METHODS: Eighty patients scheduled for flexible ureteroscopic lithotripsy were randomly divided into four groups. In groups A, B, and C, the RPPs were set at 0, - 5, and - 10 mmHg, respectively. Group D was regarded as the controls with unfixed RPP. Isotonic saline containing 1% ethanol was used as the irrigation fluid, with an average irrigation flow rate of 100 mL/min. The primary outcome of this study was the absorption of perfusion fluid that was calculated based on the exhaled ethanol concentration. The secondary outcomes included duration of operation and amounts of perfusion fluid used. Postoperative complications, pre- and postoperative renal function, infection markers, and blood gas analysis were also recorded for safety assessment. RESULTS: In all, 76 patients were involved in this study, whose demographic characteristics and preoperative conditions were comparable among groups. Under the same perfusion flow rate, the groups with fixed RPP exhibited reduced absorption of perfusion fluid, duration of operation, and perfusion volume. In particular, the lowest values were observed in group C (RPP = - 10 mmHg). In contrast to the unfixed RPP group, no considerable difference were observed in levels of BUN, Scr, WBC, CRP, and blood gas values among the fixed RPP groups. Moreover, postoperative complications showed no significant difference among groups. CONCLUSION: In flexible ureteroscopic lithotripsy, the groups with fixed RPP had less absorption of perfusion fluid and perfusion volume, shorter duration of surgery, and higher safety than the unfixed group.

[Evaluation of red and white aesthetic effects of porcelain veneers in the restoration of developmental anterior dental gaps].

PURPOSE: To evaluate the red and white aesthetic effect of porcelain veneer in the restoration of developmental anterior interdental spaces. METHODS: A total of 152 anterior teeth in 64 patients with developmental anterior dental gaps were restored using porcelain veneers, the aesthetic effects before and after restoration were evaluated by pink aesthetic index (PES) and white aesthetic index(WES), the aesthetic effect of gingival papilla filling and reconstruction was evaluated by interdental gingival papilla index (PIS), and visual analogue score (VAS) was used to compare the satisfaction of patients before and after restoration. SPSS 203.0 software package was used for statistical analysis. RESULTS: The overall mean scores of PES before and after restoration of 152 developmental anterior interdental teeth were 9.63±2.23 and 13.64±0.88, respectively. The average scores of WES before and after restoration were 6.85±1.87 and 9.81±0.58, respectively. There were significant differences of PES and WES scores before and after restoration(P＜0.01). According to the requirements of "near" perfect restoration(PES≥13 points, WES≥9 points), the red and white aesthetic effect after restoration was near the standard. The scores of PIS before and after restoration were 1.86±0.67 and 2.97±0.18, the interdental gingival papilla was completely filled with space, and the shape was ideal, there were significant statistical differences before and after restoration(P＜0.01). Patients were more satisfied with the smile curve and morphology of the restored anterior teeth than other parameters. CONCLUSIONS: The aesthetic effect of using porcelain veneers to repair developmental anterior interdental gaps is ideal, among which the filling of the papillae between the teeth can meet the expectations of patients, and the aesthetic effect evaluation of PIS and PES/WES applied to porcelain veneers to repair developmental anterior interdental spaces has practical guiding significance in clinical practice.

Insights into the phylogenetic diversity, biological activities, and biosynthetic potential of mangrove rhizosphere Actinobacteria from Hainan Island.

Mangrove rhizosphere soils host diverse Actinobacteria tolerant to numerous stresses and are inevitably capable of exhibiting excellent biological activity by producing impressive numbers of bioactive natural products, including those with potential medicinal applications. In this study, we applied an integrated strategy of combining phylogenetic diversity, biological activities, and biosynthetic gene clusters (BGCs) screening approach to investigate the biotechnological importance of Actinobacteria isolated from mangrove rhizosphere soils from Hainan Island. The actinobacterial isolates were identifified using a combination of colony morphological characteristics and 16S rRNA gene sequence analysis. Based on the results of PCR-detected BGCs screening, type I and II polyketide synthase (PKS) and non-ribosomal synthetase (NRPS) genes were detected. Crude extracts of 87 representative isolates were subjected to antimicrobial evaluation by determining the minimum inhibitory concentration of each strain against six indicator microorganisms, anticancer activities were determined on human cancer cell lines HepG2, HeLa, and HCT-116 using an MTT colorimetric assay, and immunosuppressive activities against the proliferation of Con A-induced T murine splenic lymphocytes in vitro. A total of 287 actinobacterial isolates affiliated to 10 genera in eight families of six orders were isolated from five different mangrove rhizosphere soil samples, specififically, Streptomyces (68.29%) and Micromonospora (16.03%), of which 87 representative strains were selected for phylogenetic analysis. The crude extracts of 39 isolates (44.83%) showed antimicrobial activity against at least one of the six tested indicator pathogens, especially ethyl acetate extracts of A-30 (Streptomyces parvulus), which could inhibit the growth of six microbes with MIC values reaching 7.8 µg/mL against Staphylococcus aureus and its resistant strain, compared to the clinical antibiotic ciproflfloxacin. Furthermore, 79 crude extracts (90.80%) and 48 (55.17%) of the isolates displayed anticancer and immunosuppressive activities, respectively. Besides, four rare strains exhibited potent immunosuppressive activity against the proliferation of Con A-induced T murine splenic lymphocyte in vitro with an inhibition rate over 60% at 10 µg/mL. Type I and II polyketide synthase (PKS) and non-ribosomal synthetase (NRPS) genes were detected in 49.43, 66.67, and 88.51% of the 87 Actinobacteria, respectively. Signifificantly, these strains (26 isolates, 29.89%) harbored PKS I, PKS II, and NRPS genes in their genomes. Nevertheless, their bioactivity is independent of BGCs in this study. Our findings highlighted the antimicrobial, immunosuppressive, and anticancer potential of mangrove rhizosphere Actinobacteria from Hainan Island and the biosynthetic prospects of exploiting the corresponding bioactive natural product.

[Magnetic Resonance Imaging Tracing the Biodistribution of SPIO-shRNA Molecular Probe in vivo].

OBJECTIVES: To investigate the biodistribution of superparamagnetic iron oxide (SPIO)-shRNA molecular probe by magnetic resonance imaging (MRI) in vivo. METHODS: Six New Zealand white rabbits were injected intravenously with SPIO-shRNA molecular probe (9.6 mg Fe/kg) via ear edge vein. The blood samples were collected to analyse the pharmacokinetic parameters through measuring the iron content by atomic absorption spectrometry (AAS) method at 30 min before and 1 min, 3 min, 5 min, 10 min, 15 min, 30 min and at 1, 2, 3, 6, 12, and 24 h after the injection. Six Kun Ming (KM) mice were injected intravenously with SPIO-shRNA molecular probe (4.8 mg Fe/kg). The biodistribution of SPIO-shRNA molecular probe was traced by MRI in vivo. Ninety six KM mice were randomly divided into control group and experimental group: each mouse in experimental group was injected intravenously with SPIO-shRNA molecular probe (4.8 mg Fe/kg). The liver, spleen, kidney, brain and muscle of the control group and the experimental group on 1, 3, 5, 7, 9, 11 and 14 d after the injection were collected. The organ iron content were measured by AAS method and Prussian blue staining in order to observe the distribution of the SPIO-shRNA molecular probe in the main organ. RESULTS: Our results demonstrated that the pharmacokinetics of the molecular probe complied with two-compartment model, and the blood half-life was (3.692±0.196) h. The data of MRI showed that the probe were distributed in liver and spleen, and the signs were reduced in accord with the increase of probe's doses in liver and spleen. The probe's metabolism was slow, and the probe was cleared from liver and spleen at 2 weeks after the injection. The results of AAS and Prussian blue staining further testified the results of MRI. CONCLUSIONS: Our data showed the biodistribution of SPIO-shRNA molecular probe in main organs can be traced by MRI in vivo. Meanwhile, it provides important information for the effectiveness of the probe by MRI at tumor in vivo.

[Optimal concentration of superparamagnetic iron oxide-short hairpin RNA dual functional molecular probe transfected into ovarian cancer cells in vitro].

OBJECTIVE: To explore the effects of superparamagnetic iron oxide-short hairpin RNA (SPIO-ShRNA) dual functional molecular probes of different concentrations on morphology and biological behavior of ovarian cancer SKOV3 cells in vitro. METHODS: The dual functional molecular probes at an iron concentration of 5, 15, 30, 45, 75, and 100 mg/L were transfected into SKOV3 cells. The transfection rate of the probe was observed by fluorescence microscope. The distribution and content of iron particles in SKOV3 cells were determined by Prussian blue staining, atomic adsorption spectrometer and electron microscopy. Cell viability was observed by cell counting kit-8 (CCK-8).The apoptosis was detected by flow cytometry. The expression of protein within the cells was detected by Western blot. The changes of the signal intensity were measured by magnetic resonance imaging (MRI). RESULTS: The SPIO-ShRNA dual functional molecular probe was uptaken in aconcentration-dependence manner within a certain range (5-30 mg/L). When the concentration of the probe was 45 mg/L, the labeling rate of the cell was close to 100%; With the increase of the concentration of probe, the cell survival rate decreased gradually. The cell survival rate of each experimental group were 94.626%±1.050%, 93.373%±1.180%, 91.700%±3.122%, 75.100%±4.362%, 72.983%±3.233%, 71.010%±2.910%,5, 15, 30 mg/L cell survival rate was not significantly decreased, the difference was not statistically significant (P=0.226, P=0.068, P=0.475); When the concentration of the probe was greater than or equal to 45 mg/L,the survival rate decreased obviously (P<0.001); Group of 45 mg/L protein expression rate was 68.905%±3.510%, When the concentration of the probe was greater than or equal to 45 mg/L, the inhibition rate of the protein expression level of epidermal growth factor receptor was obviously higher than those of 5, 15, and 30 mg/L groups, the difference was statistically significant (P<0.001, P=0.001, P=0.003, all P<0.01); the MRI displayed that the signal intensity was decreased with increasing concentrations of the probe. The signal intensity of 45 mg/L group was 165.55±4.92, compared with the blank control group(same volume of phosphate buffer saline), normal group (unlabeled ovarian cancer SKOV3 cells), 5, 15, and 30 mg/L groups , the signal intensity of 45 mg/L group decreased significantly (all P<0.001). CONCLUSION: The dual functional molecular probe can effectively transfect and specifically inhibit the expression of SKOV3 cell lines at the iron concentration of 45 mg/L, and can also be detected by MRI. The role of diagnosis and treatment of the dual functional molecular probe has been initially confirmed.

Effects of external magnetic field on the transfection rate of SPIO-shRNADual functional molecular probe into ovarian carcinoma SKOV3 cells in vitro.

OBJECTIVE: To explore the transfection rate of SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells in external magnetic field. METHODS: Dual functional molecular probe at an iron concentration of 45 mg/L was transfected into SKOV3 cells. The cells with coexisting probe and magnetic fields were set as the intervention group,the probe-transfected cells as negative control group, and normally cultured SKOV3 without any transfection as blank control group. The transfection rate was detected by flow cytometry. Cell viability was observed by CCK-8 assay. Epidermal growth factor receptor (EGFR) expression level in SKOV3 cells was determined by real-time quantitative PCR and Western blot analysis. The signal intensity was measured by magnetic resonance imaging (MRI). RESULTS: The transfection rate of the intervention group was (79.20 ± 3.31)%, which was significantly higher than that of negative control group (P=0.001). Compared with the negative control group,the cell viability of the intervention group significantly decreased (P=0.011), protein and mRNA expression levels of EGFR in the intervention group were significantly decreased (both P<0.05). The signal intensity on T2(*)WI in the intervention group also significantly decreased (P=0.0004). CONCLUSION: The external magnetic field can improve the transfection efficiency SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells.

[Pharmacokinetics and MR imaging of SPIO-shRNA dual functional molecular probe in vivo].

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.

[Efficacy and safety of vitamin D in the treatment of idiopathic oligoasthenozoospermia].

OBJECTIVE: To explore the efficacy and safety of vitamin D (VD) in the treatment of idiopathic oligoasthenozoospermia. METHODS: This study included 86 infertile men with idiopathic oligoasthenozoospermia, who were randomized to a VD and a control group of equal number, the former given oral VD 200 IU/d and calcium 600 mg/d,qd, while the latter administered oral vitamin E 100 mg and vitamin C 100 mg, tid. After 3 months of medication, we compared the semen parameters, adverse reactions, and pregnancy rate between the two groups. RESULTS: After medication, the count of progressively motile sperm per ejaculate was increased from (9.82 ± 3.72) x 10(6) to (21.47 ± 6.52) x 10(6) ( P < 0.05) and the proportion of progressively motile sperm from (18.41 ± 9.82)% to (28.27 ± 4.47)% (P < 0.05) in the VD group. In comparison, the count of progressively motile sperm per ejaculate was elevated from (9.51 ± 6.31) x 10(6) to (12.36 ± 4.43) x 10(6) (P > 0.05) and the proportion of progressively motile sperm from (17.79 ± 5.25)% to (21.35 ± 2.41)% (P > 0.05) in the control group. Pregnancy was achieved in 7 cases (16.3%) in the VD group, but only lease (2.3%) in the control (P < 0.05). No adverse reactions were observed in either of the groups. CONCLUSION: Vitamin D, as a safe option for the treatment of idiopathic oligoasthenozoospermia, can effectively improve the semen quality, especially the progressive sperm motility of the patient.

Dynamics of lamivudine-resistant hepatitis B virus strains in patients with entecavir rescue therapy.

Entecavir rescue therapy is frequently used in patients with lamivudine-resistant hepatitis B virus (HBV) strains. The aim of this study was to investigate evolutionary patterns of HBV quasispecies during entecavir rescue therapy and evaluate their impacts on therapeutic efficacy. We enrolled 21 chronic hepatitis B patients who failed to respond to lamivudine therapy and were switched to entecavir treatment. Measurement of serum HBV DNA and sequence analysis of HBV reverse transcriptase were done up to 144 weeks. Four patients of this series showed a reversion to wild-type HBV after entecavir treatment and in three of them, a complete viral response (<2.6 log10 copies/ml) was achieved. An additional five patients developed entecavir genotypic resistance, with prior occurrence of lamivudine-resistant mutation (L180 M ± M204 V/I). A viral breakthrough was observed in four of the five patients with entecavir-resistant mutants. The remaining 12 patients of this series showed dominance of lamivudine-resistant mutants throughout the entecavir rescue therapy, and five of them achieved a complete viral response at the end of follow-up. The average HBV DNA level was significantly lower in patients with a reversion to wild-type HBV than in those without it (P < 0.05). In conclusion, reversion to wild-type HBV is a favorable indicator for response to entecavir rescue therapy in lamivudine-refractory patients with chronic hepatitis B. The presence of lamivudine-resistant mutations contributes to the development of entecavir resistance.

[Progress in home uroflowmetry in evaluation of lower urinary tract symptoms in patients].

Uroflowmetry is the most widely used non-invasive urodynamic tool for most patients with suspected lower urinary tract dysfunction. Home uroflowmetry can produce multiple representative flow readings in adequate privacy condition when patients feel a normal desire. Because of its advantages of decreasing variability, describing circadian rhythms and combating problems inherent to clinic-based measurements, it can improve diagnostic accuracy and predictive value. Further studies are required to confirm quantitative data of patient's preferences and to measure cost-benefit of home uroflowmetry.

[Comparison of new portable home electronic uroflowmeter with Laborie uroflowmeter].

OBJECTIVE: To design a new portable home electronic uroflowmeter and compare it with traditional methods. METHODS: The system consists of collectors, urine conducting apparatus, intelligent cell phone, wireless network communication technology, computer analysis and drawing, and data storage technology, etc., and can automatically collect voiding information from patients with lower urinary tract symptoms(LUTS) Through Bluetooth, the voiding information was sent to the patient's intelligent cell phone from the collector, then stored directly by intelligent cell phone and wirelessly transmitted to the workstation in hospital. The system was primarily tested with regard to accuracy of measurement of the voided volume. Multiple doses with known volume were introduced in the system and Laborie uroflowmeter. Furthermore, 38 outpatients who had LUTS were tested simultaneously with the system and Laborie uroflowmeter. The statistical method for assessing agreement between the two methods of clinical measurement was Bland-Altman analysis. Among the subjects, there were 22 male patients and 16 female patients, ranging from 21 to 37 years old, with an average age of 25.5 years, of whom, 19 were tested once and 19 patients twice, equaling to 57 tests. RESULTS: The system could accurately collect and analyze voiding time, uroflowmetry, voided volume, and automatically provide uroflowmetry parameters. The measurement error of 100, 200, 300, 500 and 800 mL is less than 5%. 12.28%, 5.26% and 3.51% of the Qmax, Qave and voided volume points were beyond the 95% limits of agreement. The maximum absolute values of the Qmax, Qave and voided volume difference were 0.38 mL/s, 0.70 mL/s and 2.90 mL, respectively.They agreed with the recommendation of Standardization International Continence Society. CONCLUSION: The new portable home electronic uroflowmeter has good agreement with Laborie uroflowmeter,and is a new LUTS monitoring system integrated with correct, reliable, real-time, convenient and easy-managing advantages. It is as noninvasive and reliable as traditional methods, and its portable feature facilitates application out of hospitals. It can also record voiding diaries.

[Effect of polymerization-shrinkage of different resin composites curing by two different light curing units].

PURPOSE: To investigate the effect of light-emitting diode (LED) light curing unit (LCU) and quartz-tungsten (QTH) LCU on the polymerization-shrinkage of different resin composites. METHODS: Three composites Z100, Z250, AP-X were irradiated by QTH LCU and LED LCU. Polymerization-shrinkage of 50s and 400s after the composites curing were measured by 3D-profile measuring apparatus by phase-shifted projected grating.The polymerization-shrinkage of LED and QTH curing composites were analysed by SPSS12.0 software package of one-way ANOVA. RESULTS: At 50s curing and 400s curing period, the polymerization- shrinkage of Z100, Z250 resin was different by QTH LCU and LED LCU curing, the polymerization-shrinkage by LED LCU curing was less than that by QTH LCU (P < 0.05); the polymerization-shrinkage of AP-X was not statistically different by QTH LCU and LED LCU curing (P > 0.05). CONCLUSUINS: The polymerization-shrinkage of Z100,Z250 resin is statistically less cured by LED LCU than QTH; the polymerization-shrinkage of AP-X resin is not statistically different cured by QTH LCU and LED LCU. Supported by Research Fund of Science and Technology Commission of Shanghai Municipality (08DZ2271100).