目的：探讨罗伊氏乳杆菌对轮状病毒（RV） SA11株体内外复制的抑制作用，并阐明其对相关免疫因子表达的影响。方法：体外实验，培养并鉴定罗伊氏乳杆菌，绘制罗伊氏乳杆菌标准曲线和生长曲线，筛选罗伊氏乳杆菌培养最佳时间和最适浓度。采用5×10～8、10×10～8、50×10～8、100×10～8、200×10～8和500×10～8 CFU·mL^-1罗伊氏乳杆菌感染细胞，台盼蓝染色法检测Caco-2细胞存活率。将不同浓度罗伊氏乳杆菌与RV体外共孵育并作用于Caco-2细胞，Caco-2细胞分为阴性对照组（NC组）、阳性对照组（PC组）和10～7、10～8、10～9及10¹⁰ CFU·mL^-1罗伊氏乳杆菌组，免疫荧光灶法检测罗伊氏乳杆菌作用后Caco-2细胞中病毒滴度，实时荧光定量PCR （RT-qPCR）法检测不同浓度罗伊氏乳杆菌作用后Caco-2细胞中RV VP6基因拷贝数。体内实验，将25窝SPF级乳鼠分为对照组、RV组（感染SA11毒株）、Ab-NC组（抗生素处理耗竭肠道菌群）、Ab-RV组（耗竭肠道菌群后感染SA11毒株）和Ab-Lac-RV组（耗竭肠道菌群，并感染罗伊氏乳杆菌后感染SA11毒株）。收取各组乳鼠灌胃第2、4、6、8和10天的粪便样本和灌胃第4天结肠组织样本，RT-qPCR法检测各组乳鼠粪便中RV VP6基因拷贝数和结肠组织中白细胞介素（IL）-1β、IL-8、IL-10、γ干扰素（IFN-γ）和肿瘤坏死因子（TNF）-αmRNA表达水平。结果：罗伊氏乳杆菌生长良好，形态圆润，呈圆形、光滑和乳白色的凸起样菌落，且边缘较整齐；革兰染色后菌体呈紫色、不规则和方形杆状；经16SrDNA测序后序列同源性为99%，提示罗伊氏乳杆菌活化成功；罗伊氏乳杆菌活菌数与吸光度（A）值呈线性关系，回归分析标准曲线为Y=0.437 5X+0.000 6,R2=0.999 4。培养0～2 h，细菌处于对数生长期，细菌生长迟缓；培养2～14 h，细菌快速生长，并于培养14～16 h时细菌生长趋于稳定，培养16 h时达到细菌生长速率顶峰，随后进入衰亡期。5×10～8、10×10～8、50×10～8、100×10～8和200×10～8 CFU·mL^-1罗氏乳杆菌感染后，Caco-2细胞存活率均>90%，因此选用上述浓度罗氏乳杆菌感染细胞。与PC组比较，5×10～8、10×10～8、50×10～8、100×10～8和200×10～8 CFU·mL^-1罗氏乳杆菌组Caco-2细胞中RV VP6基因拷贝数均明显降低（P<0.01）。与PC组比较，10～7、10～8、10～9和10¹⁰ CFU·mL^-1罗伊氏乳杆菌组Caco-2细胞中病毒滴度均明显降低（P<0.01）。与对照组比较，Ab-NC组、Ab-RV组和Ab-Lac-RV组乳鼠肠道菌群菌落数量均明显减少，乳鼠肠道菌群耗竭成功。灌胃第2和4天，与RV组比较，Ab-RV组乳鼠粪便中RV VP6基因拷贝数明显降低（P<0.05）；灌胃第4、6、8和10天，与Ab-RV组比较，Ab-Lac-RV组乳鼠粪便中RV VP6基因拷贝数明显降低（P<0.05）。与对照组比较，Ab-RV组和Ab-Lac-RV组乳鼠结肠组织中IL-1β、IL-10、IFN-γ及TNF-α mRNA表达水平均明显升高（P<0.05或P<0.01）,IL-8 mRNA表达水平均明显降低（P<0.05）,Ab-Lac-RV组乳鼠结肠组织中IL-10 mRNA表达水平明显升高（P<0.01）。结论：罗伊氏乳杆菌可能通过上调IL-1β、IL-10、IFN-γ和TNF-α mRNA表达及下调IL-8 mRNA表达，抑制RV复制。

目的：探讨小白菊内酯（PTL）通过调控机械敏感性离子通道蛋白1 （Piezo1）表达对机械牵张应力作用下软骨细胞凋亡的影响，并阐明其相关作用机制。方法：按照拉伸性变量将软骨细胞分为0%、5%、10%、15%和20%拉伸组。另取软骨细胞，分为对照组、20%拉伸组、20%拉伸+5μmol·L^-1 PTL组、20%拉伸+10μmol·L^-1 PTL组和20%拉伸+20μmol·L^-1 PTL组。使用Piezo1短发夹RNA （shRNA）干扰慢病毒（sh-Piezo1）或shRNA-NC慢病毒感染软骨细胞，将软骨细胞分为sh-Piezo1组和sh-NC组，另设置空白对照组。另将软骨细胞分为20%拉伸组、20%拉伸+PTL组、20%拉伸+sh-Piezo1组和20%拉伸+sh-Piezo1+PTL组。Hoechst 33258荧光染色观察各组细胞核形态表现，流式细胞术检测各组细胞凋亡率，分光光度法检测各组细胞中含半胱氨酸的天冬氨酸蛋白酶（Caspase）-3活性，CCK-8法检测各组细胞增殖率，Fluo-4/AM荧光探针法检测各组细胞中钙离子(Ca²⁺)水平，实时荧光定量PCR （RT-qPCR）法检测各组细胞中Piezo1 mRNA表达水平，Western blotting法检测各组细胞中Piezo1蛋白表达水平。结果：Hoechst 33258荧光染色观察，随着拉伸量逐渐增加，0%、5%、10%、15%和20%拉伸组软骨细胞细胞核呈碎块状致密浓染的软骨细胞数量逐渐增加。流式细胞术检测，与0%拉伸组比较，5%、10%、15%和20%拉伸组软骨细胞凋亡率均明显升高（P<0.01）；与对照组比较，20%拉伸组软骨细胞中细胞凋亡率明显升高（P<0.05）；与20%拉伸组比较，20%拉伸+5μmol·L^-1 PTL组、 20%拉伸+10μmol·L^-1 PTL组和20%拉伸+20μmol·L^-1 PTL组软骨细胞细胞凋亡率均明显降低（P<0.05）；与20%拉伸组比较，20%拉伸+PTL组和20%拉伸+sh-Piezo1组软骨细胞凋亡率均明显降低（P<0.05）。分光光度法检测，与0%拉伸组表示，5%、10%、15%和20%拉伸组软骨细胞中Caspase-3活性均明显升高（P<0.01）；与对照组比较，20%拉伸组软骨细胞中Caspase-3活性明显升高（P<0.05）；与20%拉伸组比较，20%拉伸+5μmol·L^-1 PTL组、20%拉伸+10μmol·L^-1 PTL组和20%拉伸+20μmol·L^-1 PTL组软骨细胞中Caspase-3活性均明显降低（P<0.05）；与20%拉伸组比较，20%拉伸+PTL组和20%拉伸+sh-Piezo1组软骨细胞中Caspase-3活性均明显降低（P<0.05）。CCK-8法检测，与0μmol·L^-1 PTL组比较，40.00、 80.00和160.00μmol·L^-1 PTL组软骨细胞增殖率均明显降低（P<0.05），提示20.00μmol·L^-1 PTL为最高无毒性作用浓度。Fluo-4/AM荧光探针法检测，与对照组比较，20%拉伸组软骨细胞中Ca²⁺水平明显升高（P<0.05）；与20%拉伸组比较，20%拉伸+5μmol·L^-1 PTL组、20%拉伸+10μmol·L^-1 PTL组和20%拉伸+20μmol·L^-1 PTL组软骨细胞中Ca²⁺水平均明显降低（P<0.05）；与20%拉伸组比较，20%拉伸+PTL组和20%拉伸+sh-Piezo1组软骨细胞中Ca²⁺水平均明显降低（P<0.05）。RT-qPCR法检测，与空白对照组和sh-NC组比较，sh-Piezo1组软骨细胞中Piezo1 mRNA表达水平明显降低（P<0.05）。Western blotting法检测，与对照组比较，20%拉伸组软骨细胞中Piezo1蛋白表达水平明显升高（P<0.05）；与20%拉伸组比较，20%拉伸+5μmol·L^-1PTL组、20%拉伸+10μmol·L^-1 PTL组和20%拉伸+20μmol·L^-1 PTL组软骨细胞中Piezo1蛋白表达水平均明显降低（P<0.05）；与空白对照组和sh-NC组比较，sh-Piezo1组软骨细胞中Piezo1蛋白表达水平明显降低（P<0.05）。结论：PTL可抑制高强度周期性机械牵张应力诱导的软骨细胞凋亡，其作用机制可能与抑制Piezo1介导Ca²⁺内流引起的细胞凋亡有关。

Objective To discuss the effect of NOD-like receptor protein 3 （NLRP3） inflammasome on the renal interstitial fibrosis in the unilateral ureteral obstruction （UUO） model rats， and to clarify its potential mechanism. Methods Thirty healthy male Wistar rats were randomly divided into sham operation group （n=6） and UUO group （n=24）. The rats in sham operation group underwent the dissection of the ureter without ligation， while the rats in UUO group were sacrificed on the 3rd， 7th， and 14th days after operation， and based on the treatment duration，the rats were divided into UUO 3 d group （n=8）， UUO 7 d group （n=8）， and UUO 14 d group （n=8）. HE staining and Masson staining were used to observe the pathomorphology of kidney tissue of the rats in various groups； reagent kits were used to detect the levels of malondialdehyde （MDA）， activities of superoxide dismutase （SOD）， and levels of hydroxyproline （HYP） in kidney tissue of the rats in various groups；immunohistochemistry method was used to detect the expression levels of α-smooth muscle actin （α-SMA） and transforming growth factor-β1 （TGF-β1） proteins in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of NLRP3 protein in kidney tissue of the rats in various groups. Results The HE staining results showed significant tubular dilation， interstitial edema， and widening， with increased infiltration of inflammatory cells， and shedding of epithelial cells was seen in parts of the tubular lumina of the rats in UUO group. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； compared with UUO 3 d group and UUO 7 d group， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. The Masson staining results showed that in UUO group， there was evident infiltration of inflammatory cells in the renal interstitium and a noticeable increase in fibrotic tissue proliferation； with the increasing of duration of UUO， some tubular structures disappeared， and the interstitial widened further with gradually increasing collagen deposition， particularly at the corticomedullary junction. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； and compared with UUO 3 d and UUO 7 d groups， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. Compared with sham operation group， the levels of MDA in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）， and the SOD activities were significantly decreased （P<0.05）. Compared with sham operation group， the levels of HYP in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）；compared with UUO 3 d group， the level of HYP in obstructed kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）. The immunohistochemistry results showed that compared with sham operation group， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significant increased （P<0.05）； compared with UUO 3 d and UUO 7 d groups， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）； compared with sham operation group，the expression levels of TGF-β1 protein in renal tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）； compared with UUO 3 d group， the expression levels of TGF-β1 protein in the tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 14 d group were significantly decreased（P<0.05）. The Western blotting results showed that compared with sham operation group， the expression levels of NLRP3 protein in kidney tissue of the rats in UUO 7 d and UUO 14 d groups were significantly increased （P<0.05）. Conclusion The NLRP3 inflammasome plays a critical role in renal fibrosis of the UUO rats，and its mechanism may be related to the increasing of oxidative stress and the increasing of expression level of TGF-β1 protein.

Objective To discuss the improvement effect of uric acid （UA） on the postoperative cognitive dysfunction （POCD） in the mice anesthetized with isoflurane for a long duration， and to clarify its possible mechanism. Methods Twenty-four healthy male C57BL/6 mice were randomly divided into blank control group，anesthesia group， and UA group, and there were eight mice in each group. The mice in UA group were injected intraperitoneally with 200 μL UA solution daily for 2 d before anesthesia. The mice in blank control group and anesthesia group were given the same volume of saline; the mice in anesthesia group and UA group were used to prepare the models of long-duration isoflurane anesthesia， while the mice in blank control group were untreated. Y-maze tests was used to detect the alternation success rate， movement distances， and movement speeds of the mice in various groups； situational fear experiment was used to detect the percentages of freezing time； Western blotting method was used to detect the expression levels of interleukin (IL)-1β， IL-10， and mature brain-derived neurotrophic factor (mBDNF) proteins in hippocampus tissue of the mice in various groups. Results The Y-maze test results showed that compared with blank control group， the alternation success rate of the mice in anesthesia group was significantly decreased (P<0.01)； compared with anesthesia group， the alternation success rate of the mice in UA group was significantly increased (P<0.01). The situational fear experiment results showed that compared with blank control group, the percentage of freezing time of the mice in anesthesia group was significantly decreased (P<0.01)； compared with anesthesia group， the percentage of freezing time of the mice in UA group was significantly increased (P<0.05). The cued memory experiment resutls showed that there were no significant differences of the percentage of freezing time of the mice between various groups (P>0.05). The Western blotting results showed that compared with blank control group， the expression level of IL-1β protein in hippocampus tissue of the mice in anesthesia group was increased (P<0.01)， while the expression levels of IL-10 and mBDNF proteins were decreased (P<0.01)； compared with anesthesia group, the expression level of IL-1β protein in hippocampus tissue of the mice in UA group was decreased (P<0.05), and the expression levels of IL-10 and mBDNF proteins were increased (P<0.05 or P<0.01). Conclusion UA can improve the POCD in the mice, and its mechnasim may be related with its anti-inflammatory activity inhibiting the central inflammation and upregulating the mBDNF protein expression.