Efficient AAV9 Purification Using a Single-Step AAV9 Magnetic Affinity Beads Isolation.

Adeno-associated viruses (AAVs) have emerged as promising tools for gene therapy due to their safety and efficacy in delivering therapeutic genes or gene editing sequences to various tissues and organs. AAV serotype 9 (AAV9), among AAV serotypes, stands out for its ability to efficiently target multiple tissues, thus holding significant potential for clinical applications. However, existing methods for purifying AAVs are cumbersome, expensive, and often yield inconsistent results. In this study, we explore a novel purification strategy utilizing Dynabeads™ CaptureSelect™ magnetic beads. The AAV9 magnetic beads capture AAV9 with high specificity and recovery between 70 and 90%, whereas the AAVX magnetic beads did not bind to the AAV9. Through continuous interaction with AAVs in solution, these beads offer enhanced clearance of genomic DNA and plasmids even in the absence of endonuclease. The beads could be regenerated at least eight times, and the used beads could be stored for up to six months and reused without a significant reduction in recovery. The potency of the AAV9-purified vectors in vivo was comparable to that of iodixanol purified vectors.

[Xiaojiang River Basin Ecological Environmental Quality Spatiotemporal Pattern and Evolutionary Trend Analysis Using GEE from 1990 to 2022].

Assessment and monitoring of the quality of the ecological environment in the area is a very important fundamental task in the development of ecological civilization in the Xiaojiang River Basin in Yunnan Province, which serves as a demonstration area for ecological restoration in the upper reaches of the Yangtze River. The Landsat remote sensing images from 1990, 1995, 2000, 2005, 2010, 2014, 2018, and 2022 were chosen, and the four indexes of greenness （NDMVI）, humidity （WET）, dryness （NDBSI）, and heat （LST） were extracted. The remote sensing ecological index （RSEI） was created using the principal component analysis method, then the spatial and temporal patterns and trends of ecological quality in the Xiaojiang River Basin between 1990 and 2022 were examined using the GEE platform, ArcGIS 10.7 platform, and Python platform, combining the analysis methods of geographic information mapping, coefficient of variation, Mann-Kendall trend test, Sen's slope estimation, and Hurst's index. The findings demonstrated that： ① the ecological quality of the study area had more obvious geographic differentiation spatially, and by 2022, the areas with excellent and good ecological quality grades were primarily distributed in the areas with better alpine vegetation cover, and those with poor ecological quality were primarily distributed in the areas of the mudslide ravines with relatively low terrain. On a time scale, the study area's RSEI index increased from 0.41 in 1990 to 0.55 in 2022, with a fluctuating overall trend of ecological quality improvement and an average increase of 0.048（10 a） -1； this progress was directly related to a number of ecological construction initiatives that have been energetically carried out, such as converting farms to forests, preventing mudslides, saving soil and water, managing heavy metal contamination, etc. ② The RSEI was more appropriate for the evaluation of ecological quality in alpine ravine areas because, in comparison to the NDVI index, the NDVMI adopted in this study was more sensitive to vegetation information in topographic undulation areas, especially in shaded areas, and could more accurately and quantitatively describe the vegetation information. ③ The RSEI in the Xiaojiang River Basin had a mean coefficient of variation of 0.202. Overall, its volatility was low, and its high volatility was mostly concentrated in the mudslide gully area along both sides of the Xiaojiang River fracture zone, where the surface was made up of bare rocks and sediment that was easily impacted by the changing of the seasons, the climate, and human activity. ④ The quality of the ecological environment in the region was significantly improving, with the rising area reaching 85.72% of the total area and the declining area accounting for approximately 10.15% of the total area. The future trend of change will be dominated by ongoing improvement and future degradation, accounting for 44.75% and 39.97%, respectively. It is important to pay close attention to areas that could potentially degrade. The findings of this study can serve as a theoretical foundation for additional ecological environmental conservation, management, and sustainable development in the Xiaojiang River Basin.

An "off-on-enhanced on" electrochemiluminescence biosensor based on resonance energy transfer and surface plasmon coupled 3D DNA walker for ultra-sensitive detection of microRNA-21.

In this study, a novel electrochemiluminescence (ECL) biosensor was developed to detect microRNA-21 (miRNA-21) with high sensitivity by leveraging the combined mechanisms of resonance energy transfer (RET) and surface plasmon coupling (SPC). Initially, the glassy carbon electrode (GCE) were coated with Cu-Zn-In-S quantum dots (CZIS QDs), known for their defect-related emission suitable for ECL sensing. Subsequently, a hairpin DNA H3 with gold nanoparticles (Au NPs) attached at the end was modified over the surface of the quantum dots. The Au NPs could effectively quench the ECL signals of CZIS QDs via RET. Further, a significant amount of report DNA was generated through the action of a 3D DNA walker. When the report DNA opened H3-Au NPs, the hairpin structure experienced a conformational change to a linear shape, increasing the gap between the CZIS QDs and the Au NPs. Consequently, the localized surface plasmon resonance ECL (LSPR-ECL) effect replaced ECL resonance energy transfer (ECL-RET). Moreover, the report DNA was released following the addition of H4-Au NPs, resulting in the formation of Au dimers and a surface plasma-coupled ECL (SPC-ECL) effect that enhanced the ECL intensity to 6.97-fold. The integration of new ECL-RET and SPC-ECL biosensor accurately quantified miRNA-21 concentrations from 10-8 M to 10-16 M with a limit of detection (LOD) of 0.08 fM, as well as successfully applied to validate human serum samples.

Wireless sequential dual light delivery for programmed PDT in vivo.

Using photodynamic therapy (PDT) to treat deep-seated cancers is limited due to inefficient delivery of photosensitizers and low tissue penetration of light. Polymeric nanocarriers are widely used for photosensitizer delivery, while the self-quenching of the encapsulated photosensitizers would impair the PDT efficacy. Furthermore, the generated short-lived reactive oxygen spieces (ROS) can hardly diffuse out of nanocarriers, resulting in low PDT efficacy. Therefore, a smart nanocarrier system which can be degraded by light, followed by photosensitizer activation can potentially overcome these limitations and enhance the PDT efficacy. A light-sensitive polymer nanocarrier encapsulating photosensitizer (RB-M) was synthesized. An implantable wireless dual wavelength microLED device which delivers the two light wavelengths sequentially was developed to programmatically control the release and activation of the loaded photosensitizer. Two transmitter coils with matching resonant frequencies allow activation of the connected LEDs to emit different wavelengths independently. Optimal irradiation time, dose, and RB-M concentration were determined using an agent-based digital simulation method. In vitro and in vivo validation experiments in an orthotopic rat liver hepatocellular carcinoma disease model confirmed that the nanocarrier rupture and sequential low dose light irradiation strategy resulted in successful PDT at reduced photosensitizer and irradiation dose, which is a clinically significant event that enhances treatment safety.

The safety and efficacy of Sotn ureteroscopy for renal and upper ureteral calculi: a prospective multicenter randomized controlled trial.

BACKGROUND: Sotn ureteroscopy is a new lithotripsy procedure developed on the basis of ureteroscopy and includes a rigid ureteral access sheath, standard mirror, lithotripsy mirror, and Sotn perfusion aspirator. Thus, we performed a prospective multicenter randomized controlled trial comparing the safety and efficacy of Sotn ureteroscopy in the treatment of renal and upper ureteral calculi. METHODS: In this study, 224 patients with renal and upper ureteral calculi were randomly divided equally into study and control groups from March 2018 to March 2022. All the patients were approved by the hospital ethics committee (proof number: ZF-2018-164-01 and ZF-2018-165-01) of the Second Affiliate Hospital of Guangzhou University of Chinese Medicine in China. The primary outcome was stone-free rate (SFR) assessed by computed tomography on the 1st day and month after treatment and operation duration. The secondary outcome was postoperative complication rate. RESULTS: In total, for upper ureteral calculi, the SFR of 1 day after operation of the Sotn ureteroscopy group was significantly higher than the rigid ureteroscopy group (83.6% vs. 60%, P=0.006). Moreover, operative time (33.7±1.80 vs. 52.9±2.73 min, P<0.005) of the Sotn ureteroscopy group was significantly lower than the rigid ureteroscopy group. Additionally, the SFR of 1 day after operation and operative time for the study group (Sotn ureteroscopy combined with flexible ureteroscopy) and the control group (flexible ureteroscopy alone) were 63.2% and 36.8% (P=0.005), 65.6±4.06 and 80.3±4.91 (P=0.023), respectively. However, there were no significant differences in the SFR of 1 month after operation, success rate of ureteral access sheath placement, and postoperative complications between the two groups (P>0.05). In subgroups with stone diameters ≥1.5 cm and stone CT values ≥1000 Hounsfield units, Sotn ureteroscopy showed more advantages in terms of the SFR of 1 day after operation. Importantly, complications such as ureteral injury, sepsis, fever, and severe hematuria were not statistically different between the two groups (P>0.05). CONCLUSIONS: For renal and upper ureteral calculi, Sotn ureteroscopy has the advantage of a higher SFR of 1 day after the operation and a shorter operative time, suggesting that the Sotn ureteroscopy may have further potential applications in clinics.

Stomatal response to VPD in C4 plants with different biochemical sub-pathways.

C4 NAD-malic enzyme (NAD-ME) species occurs in drier regions and exhibit different drought responses compared to C4 NADP-malic enzyme (NADP-ME) species. However, a physiological mechanism explaining the geographical discrepancies remains uncertain. This study examined gas exchange patterns that might explain different distributions observed between two subtypes of C4 photosynthesis. We measured the response of leaf gas exchange to vapour pressure deficit (VPD) and CO2 in plants from six distinct C4 clades having closely related NAD-ME and NADP-ME species using a Li-Cor 6400 gas exchange system. We found that NAD-ME species exhibited greater relative reductions in stomatal conductance with increases in VPD than NADP-ME species but observed no consistent subtype differences in C4 cycle activity as indicated by the initial slope of the A response to intercellular CO2 concentration. Based on these results, we hypothesise the greater response of gs to increasing VPD may enable NAD-ME plants to outperform NADP-ME plants in hot, dry environments where VPD is normally high.

Characterization of a Novel Polysaccharide Derived from Rhizospheric Paecilomyces vaniformisi and Its Mechanism for Enhancing Salinity Resistance in Rice Seedlings.

Soil salinity is an important limiting factor in agricultural production. Rhizospheric fungi can potentially enhance crop salinity tolerance, but the precise role of signaling substances is still to be systematically elucidated. A rhizospheric fungus identified as Paecilomyces vaniformisi was found to enhance the salinity tolerance of rice seedlings. In this study, a novel polysaccharide (PPL2b) was isolated from P. vaniformisi and identified as consisting of Manp, Glcp, GalpA, and Galp. In a further study, PPL2b showed significant activity in alleviating salinity stress-induced growth inhibition in rice seedlings. The results indicated that under salinity stress, PPL2b enhances seed germination, plant growth (height and biomass), and biochemical parameters (soluble sugar and protein contents). Additionally, PPL2b regulates genes such as SOS1 and SKOR to decrease K+ efflux and increase Na+ efflux. PPL2b increased the expression and activity of genes related to antioxidant enzymes and nonenzyme substances in salinity-induced oxidative stress. Further study indicated that PPL2b plays a crucial role in regulating osmotic substances, such as proline and betaine, in maintaining the osmotic balance. It also modulates plant hormones to promote rice seedling growth and enhance their tolerance to soil salinity. The variables interacted and were divided into two groups (PC1 77.39% and PC2 18.77%) based on their relative values. Therefore, these findings indicate that PPL2b from P. vaniformisi can alleviate the inhibitory effects of salinity stress on root development, osmotic adjustment, ion balance, oxidative stress balance, and growth of rice seedlings. Furthermore, it suggests that polysaccharides produced by rhizospheric fungi could be utilized to enhance crop tolerance to salinity.

Periodontal disease and the risk of prostate cancer: a meta-analysis of cohort studies

ABSTRACT Background: Periodontal disease is reportedly associated with the risk of various systemic diseases, including pancreatic and lung cancers. However, its association with prostate cancer remains inconclusive. Herein, we explored the association of periodontal disease with the risk of prostate cancer through a meta-analysis. Materials and Methods: MEDLINE, Embase, Web of Sciences and Cochrane Library databases were searched for eligible publications up to April 2020. Multivariate adjusted risk estimates with corresponding 95% confidence intervals (CIs) were extracted and calculated using random- or fixed-effect models. Results: Nine cohort studies involving 3.353 prostate cancer cases with 440.911 participants were identified and included in the meta-analysis. We found that periodontal disease significantly increased the risk of prostate cancer by 1.40-fold (hazard ratio [HR]=1.40, 95% CI: 1.16-1.70; P=0.001; I2=76.1%) compared with normal condition. Interestingly, the risk of developing prostate cancer was not significant in patients treated with periodontal therapy (HR=1.22, 95% CI: 0.86-1.73; P=0.272; I2=65.2%). The results of subgroup analyses were also consistent and significant when stratified by study design and follow-up period, whereas conflicting results were observed in periodontal disease ascertainment stratification. These findings were robust as indicated by sensitivity analyses. Conclusions: Periodontal disease was associated with the increased risk of prostate cancer, whereas no significant association was observed in patients treated with periodontal therapy. Hence, the awareness and importance for maintaining oral health should be improved, and the underlying mechanisms linking periodontal disease and prostate cancer should be fully explored in future research.

Novel semirigid ureterorenoscope with irrigation and vacuum suction system: introduction and initial experience for management of upper urinary calculi

ABSTRACT Objective: This study aims to design a novel semirigid ureterorenoscope with irrigation and vacuum suction system and a modified ureteral access sheath (UAS) named Sotn ureterorenoscope® (Sotn=ShuoTong Medical Company) to overcome the deficiencies of the current procedure and to improve the efficiency and safety of using Sotn ureterorenoscope® for treatment of upper urinary calculi. Materials and Methods: Fifty-eight patients, comprising 31 males and 27 females, were evaluated. The medical records of 58 patients with upper urinary calculi treated with Sotn ureterorenoscope® from March 2015 to June 2017 were retrospectively reviewed at the Second Affiliate Hospital of Guangzhou University of Chinese Medicine in China. The primary outcome was stone-free rate (SFR) assessed by computed tomography on the 1st day and one month after treatment. The secondary outcome was postoperative complication rate. Results: The mean and SD of operative duration was 48.5 (10.4) min, and the mean and SD of stone size was 15.6 (5.6) mm. The primary overall SFR was 89.7% (52/58) and 100% at 1 month follow-up. Complication, which was Clavien I (minor fever managed by antipyretic therapy), was detected in 1.7% (1/58) of the patients. Conclusions: Sotn ureterorenoscope® is technically feasible, efficacious and safe for treatment of upper urinary calculi because of its advantages of high SFR and low complication rates.

Risk factors associated with systemic inflammatory response syndrome after percutaneous nephrolithotomy in China:A Meta-analysis / 中国内镜杂志

Objective To explore the risk factors of systemic inflammatory response syndrome crisis (SIRS) after percutaneous nephrolithotomy (PCNL) in China. Methods Databases of CNKI, CBM, WanFan and VIP were searched to retrieve studies about systemic inflammatory response syndrome after percutaneous nephrolithotomy to October, 2016. Results 18 studies involving 5,323 patients were included. The results of meta-analysis showed that:a) univariate analysis indicated that renal insufficiency [O(R) =2.78, 95%CI (1.96 to 3.95), P = 0.000], preoperative positive urine culture [O(R) = 3.41, 95%CI (1.89 to 6.15), P = 0.000], preoperative routine urine leucocyte positive [O(R) = 3.78, 95%CI (3.02 to 4.72), P = 0.000], diabetes mellitus [O(R) = 2.14, 95%CI (1.33 to 3.45), P = 0.002], pelvic positive urine culture [O(R)= 5.14, 95%CI (2.46 to 10.73), P = 0.000] and operation time ≥120 min [O(R) = 2.31, 95%CI (1.40 to 3.82), P = 0.001] were the risk factors of SIRS; b) multivariate analysis showed that, preoperative positive urine culture [O(R) = 6.83, 95%CI (2.82 to 16.57), P = 0.000], preoperative routine urine leucocyte positive [O(R) = 5.43, 95%CI (3.51 to 8.41), P = 0.000], diabetes mellitus [O(R) = 2.85, 95%CI (1.45 to 5.58), P = 0.002], pelvic positive urine culture [O(R) = 4.30, 95%CI (1.30 to 14.21), P = 0.020] and operation time ≥120 min [O(R) = 2.72, 95%CI (1.62 to 4.59), P = 0.000] were the independent risk factors of MCAT. Conclusion The independent risk factors of SIRS for patients after PCNL are diabetes mellitus, preoperative positive urine culture, preoperative routine urine leucocyte positive, pelvic positive urine culture and operation time. However, due to the quantity and low quality of the included literature, the conclusion needs the support from high quality studies.

Risk factors associated with systemic inflammatory response syndrome after percutaneous nephrolithotomy in China:A Meta-analysis / 中国内镜杂志

Objective To explore the risk factors of systemic inflammatory response syndrome crisis (SIRS) after percutaneous nephrolithotomy (PCNL) in China. Methods Databases of CNKI, CBM, WanFan and VIP were searched to retrieve studies about systemic inflammatory response syndrome after percutaneous nephrolithotomy to October, 2016. Results 18 studies involving 5,323 patients were included. The results of meta-analysis showed that:a) univariate analysis indicated that renal insufficiency [O(R) =2.78, 95%CI (1.96 to 3.95), P = 0.000], preoperative positive urine culture [O(R) = 3.41, 95%CI (1.89 to 6.15), P = 0.000], preoperative routine urine leucocyte positive [O(R) = 3.78, 95%CI (3.02 to 4.72), P = 0.000], diabetes mellitus [O(R) = 2.14, 95%CI (1.33 to 3.45), P = 0.002], pelvic positive urine culture [O(R)= 5.14, 95%CI (2.46 to 10.73), P = 0.000] and operation time ≥120 min [O(R) = 2.31, 95%CI (1.40 to 3.82), P = 0.001] were the risk factors of SIRS; b) multivariate analysis showed that, preoperative positive urine culture [O(R) = 6.83, 95%CI (2.82 to 16.57), P = 0.000], preoperative routine urine leucocyte positive [O(R) = 5.43, 95%CI (3.51 to 8.41), P = 0.000], diabetes mellitus [O(R) = 2.85, 95%CI (1.45 to 5.58), P = 0.002], pelvic positive urine culture [O(R) = 4.30, 95%CI (1.30 to 14.21), P = 0.020] and operation time ≥120 min [O(R) = 2.72, 95%CI (1.62 to 4.59), P = 0.000] were the independent risk factors of MCAT. Conclusion The independent risk factors of SIRS for patients after PCNL are diabetes mellitus, preoperative positive urine culture, preoperative routine urine leucocyte positive, pelvic positive urine culture and operation time. However, due to the quantity and low quality of the included literature, the conclusion needs the support from high quality studies.