扶正方对Lewis肺癌小鼠免疫功能、PI3K/AKT信号通路和外周血IL-2、IL-6、INF-γ的影响

摘要 目的：观察扶正方对Lewis肺癌小鼠免疫功能、磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路和外周血白细胞介素（IL）-2、IL-6、γ干扰素（INF-γ）的影响。方法：将40只Lewis肺癌小鼠随机分为模型组（M组）、扶正方低剂量组（A组）、扶正方高剂量组（B组）、顺铂组（S组）,每组10只,A组、B组分别给予扶正方0.4 mL/20 g、0.8 mL/20 g灌胃,M组给予生理盐水0.4 mL/20 g灌胃,S组给予顺铂1 mg/mL,0.4 mL灌胃,连续14d,比较各组小鼠一般情况、肿瘤重量,胸腺指数、脾脏指数、脾脏CD3⁺细胞、CD4⁺细胞、CD8⁺细胞比例细胞百分比,鼠肿瘤组织PI3K/AKT信号通路蛋白表达水平及外周血IL-2、IL-6、INF-γ水平。结果：A组、B组、S组小鼠肿瘤重量低于M组,S组小鼠肿瘤重量低于A组、B组（P<0.05）,治疗前各组小鼠体重比较无统计学差异（P>0.05）,治疗后A组、B组小鼠体重高于S组、M组（P<0.05）。A组、B组小鼠胸腺指数显著高于M组、S组（P<0.05）。A组、B组CD3⁺、CD4⁺、CD4⁺/CD8⁺显著高于M组、S组,CD8⁺显著低于M组、S组（P<0.05）,B组CD3⁺、CD4⁺、CD4⁺/CD8⁺显著高于A组,CD8⁺低于A组（P<0.05）。A组、B组、S组小鼠肿瘤组织PI3K蛋白、AKT蛋白表达水平显著低于M组（P<0.05）。A组、B组、S组小鼠外周血IL-2、INF-γ水平显著高于M组,IL-6水平显著低于M组（P<0.05）。结论：扶正方可以提升Lewis肺癌小鼠免疫功能,调节IL-2、IL-6、INF-γ细胞因子水平,抑制PI3K/AKT信号通路起到抗肺癌的作用。     

北京市五大水系水体浮游细菌的数量特征研究

为掌握水域浮游细菌数量分布特征及其变化情况,采用荧光显微镜细菌计数法（AODC）于2016年6月至2018年9月研究了北京市五大水系浮游细菌的数量特征。结果表明,各水系浮游细菌密度分别为潮白河水系（0.24～28.46）×104 cell/mL,大清河水系（1.34～64.00）×10⁴ cell/mL,永定河水系（0.17～6.77）×10⁴ cell/mL,北运河水系（0.24～64.00）×10⁴ cell/mL,蓟运河水系（0.80～112.00）×10⁴ cell/mL。SPSS相关分析表明,各水系浮游细菌密度与水体理化因子间的相关性存在明显差异,大清河水系细菌密度与TN（P<0.05）呈显著正相关,与TP（P<0.01）和ADP(P<0.01)呈极显著正相关;永定河水系细菌密度与TAN（P<0.05）和TP(P<0.05)呈显著正相关,与ADP（P<0.01）呈极显著正相关;蓟运河水系细菌密度与Chl-a（P<0.05）呈显著正相关;潮白河水系及北运河水系细菌密度则与各理化因子均无显著相关性。从浮游细菌数量来看,各水系水质均较好,其中永定河水系水质最优。     

静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效及外周血NLR、PCT变化观察

摘要 目的：观察静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效及外周血中性粒细胞/淋巴细胞比值（NLR）、降钙素原(PCT)变化。方法：选取2011年1月~2020年1月我院收治的败血症患儿80例为研究对象,按数字随机表法分为对照组和观察组各40例,对照组给予万古霉素治疗,观察组在对照组基础上给予静注人免疫球蛋白治疗,比较两组临床疗效、症状改善时间和住院时间、NLR、PCT、超敏C反应蛋白(hs-CRP)、白细胞计数（WBC）、免疫功能及不良反应发生率。结果：观察组治疗有效率高于对照组（87.50%vs65.00%）（P<0.05）。观察组神经系统症状改善时间、体温改善时间、拒奶改善时间和住院时间为（6.22±1.05）d、（3.88±0.25）d、（5.10±0.86）d、（8.71±2.05）d,均短于对照组的（8.76±1.53）d、（6.22±0.64）d、（7.53±1.46）d和（11.24±3.36）d,比较差异有统计学意义（P<0.05）。治疗后观察组外周血NLR、PCT、hs-CRP、WBC水平为（1.35±0.20）、（0.80±0.34）mg/mL、（3.56±0.62）g/L、（9.12±1.80）×10⁹/L,均显著低于对照组的（1.83±0.32）、（2.19±0.73）mg/mL、（9.78±2.64）g/L和（12.26±2.59）×10⁹/L,比较差异有统计学意义（P<0.05）。治疗后观察组CD4⁺、CD3⁺、CD4/CD8为（42.77±11.36）％、（41.27±11.26）％、（1.70±0.33）,均显著高于对照组的（35.80±9.32）％、（35.66±9.40）％和（1.29±0.25）,比较差异有统计学意义（P<0.05）。两组不良反应发生率比较无差异（10.00%vs7.50%）（P>0.05）。结论：静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效显著,可降低炎症因子,提高免疫功能,且安全性较高。     

丹参酮结合有氧运动对高血压大鼠血管功能的影响及作用机制分析

摘要 目的：分析丹参酮ⅡA（TIIA）结合有氧运动（AE）对高血压大鼠血管功能的影响及作用机制。方法：选择30只自发性高血压大鼠（SHR）,随机分为模型组、TⅡA组、TⅡA+AE组,每组10只,另选取10只健康大鼠作为对照组。TⅡA组大鼠给予20 mg/kg的TⅡA,TⅡA+AE组在此基础上进行中等强度的有氧运动,模型组和健康对照组大鼠均灌胃给予等体积的生理盐水溶液。观察各组大鼠收缩压（SBP）和舒张压（DBP）的变化,HE染色观察胸主动脉组织形态学变化,测定血管舒缩功能,检测血清血管紧张素Ⅱ（AngⅡ）、内皮素-1（ET-1）和一氧化氮（NO）的表达水平,测定胸主动脉中腺苷酸活化蛋白激酶（AMPK）、核因子E2相关因子（Nrf2）、血红素加氧酶1（HO-1）、醌氧化还原酶1（NQO-1）蛋白的相对表达。结果：TⅡA组和TⅡA+AE组大鼠的SBP和DBP均低于同期的模型组大鼠;TⅡA+AE组大鼠的SBP和DBP在第8周时均低于同期TⅡA组大鼠（P<0.05）;与模型组相比,在1×10^-7、1×10^-6和1×10^-5mol/L的PE浓度下TⅡA组和TⅡA+AE组大鼠胸主动脉血管环的收缩率均降低,且TⅡA+AE组大鼠低于同浓度下的TⅡA组（P<0.05）;在1×10^-8、1×10^-7和1×10^-6的Ach浓度下TⅡA组和TⅡA+AE组大鼠胸主动脉血管环的舒张率高于模型组,且ⅡA+AE组大鼠高于同浓度下的TⅡA组（P<0.05）;TⅡA组和TⅡA+AE组大鼠血清AngⅡ和ET-1的表达水平低于模型组、NO的表达水平高于模型组（P<0.05）;与TⅡA组相比,TⅡA+AE组大鼠血清AngⅡ和ET-1的表达水平较低、NO的表达水平较高（P<0.05）;TⅡA组和TⅡA+AE组大鼠胸主动脉中AMPK、Nrf2、HO-1、和NQO-1蛋白的相对表达显于模型组,且TⅡA+AE组高于TⅡA组（P<0.05）。结论：TⅡA联合有氧运动可降低SHR的血压,改善血管舒缩功能和内皮功能,其机制可能与AMPK/Nrf2信号通路有关。     

青霉素钠对萼花臂尾轮虫适合度的促进作用及其与藻密度的关系

以萼花臂尾轮虫（Brachionus calyciflorus）为受试动物,在1.0×10⁶（较低）、2.0×10⁶（中等）和4.0×10⁶个细胞/mL （较高）的斜生栅藻（Scenedesmus obliquus）密度下,研究了不同浓度（0、200、400、600、800和1000 μg/L）的青霉素钠对轮虫各生命表统计学参数的影响。结果表明,与各藻密度下的对照组相比,当藻密度为2.0×10⁶个细胞/mL时,200-1000 μg/L的青霉素钠处理组中轮虫的生命期望和世代时间分别延长了40.67%-70.67%和34.04%-50.23%（P<0.05）,净生殖率和内禀增长率分别提高了204.35%-358.70%和36.26%-62.09%（P<0.05）;藻密度的降低或升高均显著降低青霉素钠对轮虫存活、生殖和种群增长的促进幅度。1.0×10⁶ 个细胞/mL的斜生栅藻密度下,400 μg/L的青霉素钠处理组中轮虫的生命期望延长了21%（P<0.05）,200-400μg/L的青霉素钠处理组中轮虫的世代时间延长了23.15%-33.13%（P<0.05）,200-600和1000 μg/L的青霉素钠处理组中轮虫的净生殖率分别升高了40%-81.05%和41.05%（P<0.05）;但4.0×10⁶个细胞/mL的斜生栅藻密度下,各青霉素钠处理组中轮虫的所有生命表统计学参数均与对照组之间无显著性差异（P>0.05）。藻密度对轮虫的生命期望、净生殖率、内禀增长率和后代混交率均具有显著性影响（P<0.01）,青霉素钠浓度对轮虫的世代时间、净生殖率和内禀增长率均具有显著性影响（P<0.05）,藻密度和青霉素钠浓度的交互作用对轮虫的生命期望、净生殖率和内禀增长率也具有显著性影响（P<0.05）。在实验设置的青霉素钠浓度范围内,1.0×10⁶个细胞/mL藻密度下轮虫的生命期望、世代时间和净生殖率与青霉素钠浓度之间均具有显著的剂量-效应关系（P<0.05）;2.0×10⁶个细胞/mL藻密度下,轮虫的生命期望、世代时间和内禀增长率与青霉素钠浓度之间均具有显著的剂量-效应关系（P<0.05）。本研究结果提示,环境相关浓度的青霉素钠不会对萼花臂尾轮虫的存活、生殖和种群增长产生显著性影响。     

白细胞、C反应蛋白与凝血指标在哮喘发作期患儿的临床意义

摘要 目的：探讨白细胞（WBC）、C反应蛋白（CRP）与常见凝血指标凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）、纤维蛋白原（FIB）、凝血酶时间（TT）、抗凝血酶Ⅲ（ATⅢ）、D二聚体（D-D）在哮喘发作期儿童的临床意义。方法：选取发作期哮喘儿童,根据发作严重程度分为轻度、中度、重度3组。比较指标的组间差异,Pearson相关计算WBC、CRP与凝血指标的相关性,Logistic评估中度或重度哮喘发作期的影响因素。结果：全血WBC在中度组（12.02 ×10⁹ /L ± 4.61 ×10⁹ /L）显著高于轻度组（9.56 ×10⁹ /L ± 3.21 ×10⁹ /L,P < 0.05）,重度组（12.91×10⁹ /L ± 3.14 ×10⁹ /L）显著高于轻度组（9.56 ×10⁹ /L ± 3.21 ×10⁹ /L,P < 0.05）;重度组抗凝血酶ATⅢ（109% ± 13%）高于轻度组（99% ± 13%,P < 0.05）。WBC与FIB正相关（r = 0.297, P = 0.018）,CRP与TT（r = -0.330, P = 0.008）、ATⅢ（r = -0.375, P = 0.002）负相关,与FIB（r = 0.496, P = 0.001）、D-D（r = 0.326, P = 0.009）正相关。Logistic回归显示校正性别年龄的WBC影响重度组的比值比（OR）为1.602。结论：WBC和儿童哮喘发作严重程度有关,哮喘发作期儿童体内炎症状态与凝血指标存在一定关联。     

钙离子浓度对脂肪干细胞增殖和成肌分化的影响

摘要 目的：本研究旨在探讨钙离子浓度对脂肪干细胞（ADSCs）增殖和成肌分化的影响。方法：利用抽脂手术废弃脂肪提取脂肪干细胞,进行体外细胞培养和鉴定。设立正常细胞培养液为对照组,在培养液中添加不同浓度的钙离子（分别为0.001 mol/mL、0.002 mol/mL、0.003 mol/mL、0.004 mol/mL、0.005 mol/mL共5组）为实验组,测定在48 h和96 h条件下不同浓度钙离子(Ca²⁺)对cck8反应细胞增殖的影响。同时通过TGF-β诱导成肌分化,测定不同钙离子浓度下Ca²⁺信号相关蛋白的表达,并通过CCK-8检测评价其分化水平。另外通过免疫荧光和PCR检测细胞表面特异性抗原(α-SMA,SMMHC)的表达水平。各组间比较应用独立样本 t 检验,P<0.05为差异具有统计学意义。结果：显微镜下观察可见原代ADSCs细胞培养0.5 天后贴壁生长,形态呈圆或椭圆形,而TGF-β诱导ADSCs成肌分化可见细胞呈梭形;实验组中,钙浓度<0.002 mmol/mL时,脂肪干细胞增殖得到显著促进（t=10.08, df=10, P<0.05）;钙浓度<0.003 mmol/mL时,脂肪干细胞的成肌分化也显著增加,同时免疫荧光和PCR检测发现低浓度下α-SMA,SMMHC的表达均显著增加（P<0.05）,而高钙浓度下两者均受到抑制（P<0.05）。结论：低钙浓度(钙浓度<0.002 mmol/mL)可显著促进脂肪干细胞增殖和成肌分化,而高钙浓度下两者均受到抑制。     

兔前交叉韧带损伤后膝关节本体感觉、残端血运及膝关节腔尿激酶型纤溶酶原激活物变化的实验研究

摘要 目的：研究兔前交叉韧带（ACL）损伤后膝关节本体感觉、残端血运及膝关节腔尿激酶型纤溶酶原激活物（u-PA）的变化情况。方法：选取80只新西兰兔进行研究,将其以随机数字表法分作模型组及空白对照组各40只。模型组建立单侧ACL损伤模型,空白对照组仅切开关节。比较两组术前及术后2周、4周、8周时膝关节本体感觉、残端血运及膝关节腔u-PA水平的差异。结果：模型组新西兰兔术后2周、4周、8周时的体感诱发电位（SEP）、肌电图（EMG）潜伏期均高于空白对照组,而SEP、EMG波幅均低于空白对照组（P＜0.05）。模型组新西兰兔术后2周、4周、8周时的残端组织微血管密度分别为（2.04±0.24）n/mm²、（2.75±0.61）n/mm²、（1.60±0.33）n/mm²,均高于空白对照组的（1.34±0.24）n/mm²、（1.34±0.25）n/mm²、（1.35±0.26）n/mm²,差异均有统计学意义（P＜0.05）。模型组新西兰兔术后2周、4周、8周时的膝关节液u-PA水平分别为（173.97±14.29）pg/mL、（188.37±15.82）pg/mL、（171.38±14.76）pg/mL,均高于空白对照组的（158.02±10.18）pg/mL、（157.68±10.20）pg/mL、（157.37±10.07）pg/mL,差异均有统计学意义（P＜0.05）。结论：ACL损伤后会在不同程度上影响膝关节本体感觉、残端血运及膝关节腔u-PA含量,值得临床进一步研究。     

长期肾移植受者外周血B淋巴细胞亚群分布特征

摘要 目的：观察移植肾功能稳定的长期受者（>10年）外周血B细胞亚群分布特征及其相关因素。方法：54名肾移植受者接受流式细胞仪检查,测算外周血总B细胞、未转化记忆B细胞、转化记忆B细胞、双阴性B细胞（CD19⁺CD27^-IgD^-）比例及数量（个/微升）。患者均服用包括环孢霉素的免疫抑制治疗。术后时间16.33±5.98年,GFR：91.63±11.28 mL/min/1.73 m²。结果：1长期肾移植患者外周血B细胞中幼稚B细胞最多（37.92% ± 22.06%）,未转化记忆B细胞最少(16.23% ± 11.10%)。B细胞亚群数量与白细胞总数、中性粒细胞比例等相关。2 以上述条件为控制因素行相关分析,转化记忆B细胞比例和GFR相关（r=-0.279,P=0.045）,双阴性B细胞数量和环孢霉素浓度相关（r=-0.300,P=0.029）。线性回归显示双阴性B细胞数目与环孢霉素浓度相关（R²=0.123,P=0.049）。3按GFR将患者分为肾功能减退组（GFR<90 mL/min/1.73 m²,n=19）和肾功能正常组（GFR≥90 mL/min/1.73 m²,n=35）。前者转化记忆B细胞比例显著升高（23.61% ± 10.96% vs.17.48%±8.91%,P=0.030）。按环孢霉素谷浓度将患者分为低浓度组（<64 mmol/L,n=28）和高浓度组（≥64 mmol/L,n=26）,前者双阴性B细胞数量显著升高（13.74±10.70 vs. 8.14±6.72/μL,P=0.027）。转化记忆B细胞比例与GFR分组相关（r=-0.326,P=0.018）,双阴性B细胞数量和环孢霉素浓度分组相关（r=-0.350,P=0.01）。结论：移植肾功能稳定的长期存活受者（>10年）外周血幼稚B细胞较多。转化记忆B细胞增多与移植肾功能减退相关,增多的双阴性B细胞和低孢霉素浓度治疗相关。     

川续断提取皂甙促进骨质疏松模型中骨髓基质细胞成骨分化的作用机制研究

摘要 目的：探讨川续断提取皂甙（ASA）促进骨质疏松模型中骨髓基质细胞（rBMSCs）成骨分化的作用机制。方法：选取3月龄的雌性SD大鼠60只,随机分为卵巢切除组和假手术组,每组30只。采取卵巢切除法构建骨质疏松模型。建模成功后采用微型计算机断层扫描获得其松质骨微观结构的三维图像并进行分析。采用CCK-8法测定ASA对卵巢切除组rBMSCs增殖的影响。分析碱性磷酸酶（ALP）活性;荧光定量PCR检测成骨基因ALP、骨桥蛋白（OPN）、Runt相关转录因子2（RUNX2）的表达情况;蛋白免疫印迹试验检测磷脂酰肌醇3激酶（PI3K）、磷酸化蛋白激酶B（p-AKT）蛋白表达情况。结果：与假手术组比较,卵巢切除组骨小梁数量、骨小梁厚度和骨体积分数下降,但骨小梁分离度升高,差异有统计学意义（P＜0.05）。第4天和第7天,ASA(10^-5 mol/L)组、ASA(10^-6 mol/L) 组、ASA(10^-7 mol/L) 组和ASA(10^-8 mol/L) 组rBMSC增殖均显著高于ASA(0学艺术mol/L)组,以ASA(10^-5 mol/L)最为显著;而ASA(10^-1 mol/L) 组、ASA(10^-2 mol/L) 组、ASA(10^-3 mol/L) 组和ASA(10^-4 mol/L) 组rBMSC增殖显著低于ASA(0 mol/L)组,以ASA(10^-4 mol/L)最为显著,差异均有统计学意义（P＜0.05）。第7天和第14天,ASA(10^-5 mol/L)组、ASA(10^-6 mol/L) 组、ASA(10 ^-7 mol/L) 组和ASA(10^-8 mol/L) 组ALP活性均显著高于ASA(0 mol/L)组,且第14天ALP活性高于第7天,差异均有统计学意义（P＜0.05）。与对照组比较,ASA组成骨相关基因ALP、OPN和RUNX2相对mRNA表达水平显著升高,差异有统计学意义（P＜0.05）;与ASA组比较,wortmannin组成骨相关基因ALP、OPN和RUNX2相对mRNA表达水平均显著降低,差异有统计学意义（P＜0.05）。与对照组比较,ASA组PI3K、p-AKT蛋白表达水平显著升高,差异有统计学意义（P＜0.05）;与ASA组比较,wortmannin组PI3K、p-AKT蛋白表达水平显著降低,差异均有统计学意义（P＜0.05）。结论：ASA能促进骨质疏松模型rBMSCs增殖,增强ALP活性,增强ALP、OPN和RUNX2的表达,而PI3K通路抑制剂wortmannin降低了这些成骨作用,并降低了ASA诱导的PI3K、p-AKT水平,表明ASA通过PI3K/AKT信号通路促进骨质疏松模型rBMSCs成骨分化。     

p75NTR optimizes the osteogenic potential of human periodontal ligament stem cells by up‐regulating α1 integrin expression

Human periodontal ligament stem cells (hPDLSCs) are a promising source in regenerative medicine. Due to the complexity and heterogeneity of hPDLSCs, it is critical to isolate homogeneous hPDLSCs with high regenerative potential. In this study, p75 neurotrophin receptor (p75NTR) was used to isolate p75NTR⁺ and p75NTR^? hPDLSCs by fluorescence‐activated cell sorting. Differences in osteogenic differentiation among p75NTR⁺, p75NTR^? and unsorted hPDLSCs were observed. Differential gene expression profiles between p75NTR⁺ and p75NTR^? hPDLSCs were analysed by RNA sequencing. α1 Integrin (ITGA1) small interfering RNA and ITGA1‐overexpressing adenovirus were used to transfect p75NTR⁺ and p75NTR^? hPDLSCs. The results showed that p75NTR⁺ hPDLSCs demonstrated superior osteogenic capacity than p75NTR^? and unsorted hPDLSCs. Differentially expressed genes between p75NTR⁺ and p75NTR^? hPDLSCs were highly involved in the extracellular matrix‐receptor interaction signalling pathway, and p75NTR⁺ hPDLSCs expressed higher ITGA1 levels than p75NTR^? hPDLSCs. ITGA1 silencing inhibited the osteogenic differentiation of p75NTR⁺ hPDLSCs, while ITGA1 overexpression enhanced the osteogenic differentiation of p75NTR^? hPDLSCs . These findings indicate that p75NTR optimizes the osteogenic potential of hPDLSCs by up‐regulating ITGA1 expression, suggesting that p75NTR can be used as a novel cell surface marker to identify and purify hPDLSCs to promote their applications in regenerative medicine.     

不同剂量当归口服对硫化钠致小鼠斑秃模型的影响

目的:探讨当归口服对硫化钠致小鼠斑秃模型的影响及其可能机制。方法:将50只雄性KM小鼠随机均分为空白对照组、模型组和当归高、中、低剂量组,每组10只。模型组和当归高、中、低剂量组均以8%硫化钠酒精溶液局部外涂建立斑秃模型,当归高、中、低剂量组予以不同剂量当归煎剂灌胃,空白对照组和模型组予以等体积生理盐水灌胃,每天1次,连续7 d。采用酶联免疫吸附(ELISA)和苏木-伊红素(HE)染色法分别检测小鼠血清白细胞介素-10(IL-10)水平和单位视野毛囊数,同时对小鼠背部脱毛区毛发生长情况进行评分。结果:模型组毛发生长评分、单位视野毛囊数及血清IL-10水平均显著低于空白对照组(P0.05)。与模型组比较,高、中、低剂量组毛发生长评分、单位视野毛囊数及血清IL-10水平均显著高于模型组(P0.05)。中剂量组组毛发生长评分明显高于高、低剂量组(P0.05),低剂量组评分与高剂量组毛发生长评分、单位视野毛囊数及血清IL-10水平比较无明显差异(P0.05)。结论:口服当归对硫化钠致斑秃小鼠模型有生发作用,且以中剂量效果最佳,其生发作用可能与提高血清IL-10水平有关。     

自拟益气活血方治疗小儿脾肾气虚型肾病综合征的疗效观察

目的:探讨自拟益气活血方治疗小儿脾肾气虚型肾病综合征的临床疗效。方法:选择2014年6月到2016年10月我院收治的80例脾肾气虚型肾病综合征患儿,按随机数字表法分为对照组和治疗组各40例。对照组给予强的松治疗,治疗组在对照组治疗的基础上给予自拟益气活血方治疗,两组均治疗4个月。评估两组临床疗效,检测治疗前后两组24h尿蛋白、总胆固醇(TC)、血浆白蛋白(Alb)以及肾功能指标包括尿素氮(BUN)、血肌酐(Scr)、血肌酐清除率(Ccr)。结果:治疗组的总有效率为95.00%,明显高于对照组的67.50%,差异具有统计学意义(P0.05)。治疗后,两组24 h尿蛋白、TC、BUN及Scr水平低于治疗前,Alb、Ccr水平高于治疗前,且治疗组上述各指标水平变化均显著优于对照组,差异均具有统计学意义(P0.05)。结论:自拟益气活血方治疗小儿脾肾气虚型肾病综合征的临床疗效显著,能够明显改善患儿肾功能,值得在临床上推广应用。     

Oestrogen retains human periodontal ligament stem cells stemness in long‐term culture

Objectives

During long‐term culture, loss of stemness is observed which greatly restricts the application of human periodontal ligament stem cells (hPDLSCs) in tissue regeneration. Oestrogen (E2) was found to significantly enhance the proliferation and osteogenic differentiation capacity in mesenchymal stem cells. Therefore, in this study, we investigated effects of E2 on hPDLSCs stemness in long‐term culture.

Materials and methods

Effects of E2 on hPDLSCs stemness were systematically evaluated. To characterize underlying the mechanisms, its effects on PI3K/AKT signalling pathway were determined.

Results

Our results showed that E2 was able to enhance the proliferation, modify cell cycle, up‐regulate stemness‐related genes expression, promote osteogenic differentiation and elevate the positive rate of CD146 and STRO‐1 over 10 passages in hPDLSCs. Importantly, PI3K/AKT signing pathway might play a role in these effects.

Conclusions

These findings suggest that E2 retains hPDLSCs stemness in long‐term culture, which might enhance its application in tissue engineering.     

Feasibility and cost analysis of day 4 granulocyte colony-stimulating factor mobilized peripheral blood progenitor cell collection from HLA-matched sibling donors

BackgroundGuidelines recommend treatment with 4–5 days of granulocyte colony-stimulating factor (G-CSF) for optimal donor peripheral blood progenitor cell (PBPC) mobilization followed by day 5 collection. Given that some autologous transplant recipients achieve adequate collection by day 4 and the possibility that some allogeneic donors may maximally mobilize PBPC before day 5, a feasibility study was performed evaluating day 4 allogeneic PBPC collection.MethodsHLA-matched sibling donors underwent collection on day 4 of G-CSF for peripheral blood (PB) CD34⁺ counts ≥0.04 × 10⁶/mL, otherwise they underwent collection on day 5. Those with inadequate collected CD34⁺ cells/kg recipient weight underwent repeat collection over 2 days. Transplant and PBPC characteristics and cost analysis were compared with a historical cohort collected on day 5 per our prior institutional algorithm.ResultsOf the 101 patient/donor pairs, 50 (49.5%) had adequate PBPC collection on day 4, with a median PB CD34⁺ cell count of 0.06 × 10⁶/mL. Day 4 donors were more likely to develop bone pain and require analgesics. Median collected CD34⁺ count was significantly greater, whereas total nucleated, mononuclear and CD3⁺ cell counts were significantly lower, at time of transplant infusion for day 4 versus other collection cohorts. There were no significant differences in engraftment or graft-versus-host disease. Cost analysis revealed 6.7% direct cost savings for day 4 versus historical day 5 collection.DiscussionDay 4 PB CD34⁺ threshold of ≥0.04 × 10⁶/mL identified donors with high likelihood of adequate PBPC collection. Day 4 may be the optimal day of collection for healthy donors, without adverse effect on recipient transplant outcomes and with expected cost savings.     

Mobilization and engraftment of peripheral blood stem cells in healthy related donors >55 years old

Background aimsThe increasing scarcity of young related donors has led to the use of older donors for related allogeneic hematopoietic stem cell transplantation (HSCT). This study analyzed the influence of age on the results of mobilization of peripheral blood stem cells (PBSCs) in healthy donors as well as on the engraftment and outcome of HSCT.MethodsA retrospective analysis from a single center was performed comparing the results of PBSC mobilization from related healthy donors according to their age.ResultsThe study included 133 consecutive related donors. The median age was 50 years (range, 4–77 years); 70 (53%) donors were males, and 44 (33%) were >55 years old. All donors were mobilized with granulocyte colony-stimulating factor for 5 days. The peak CD34⁺ cell count in peripheral blood was higher in younger than in older donors (median, 90.5 CD34⁺ cells/μL [range, 18–240 CD34⁺ cells/μL] versus 72 CD34⁺ cells/μL [range, 20–172.5 CD34⁺ cells/μL], P = 0.008). The volume processed was lower in younger than in older donors (16,131 mL [range, 4424–36,906 mL] versus 18,653 mL [range, 10,003–26,261 mL], P = 0.002) with similar CD34⁺ cells collected (579.3 × 10⁶ cells [range, 135.14 × 10⁶–1557.24 × 10⁶ cells] versus 513.69 × 10⁶ cells [range, 149.81 × 10⁶–1290 × 10⁶ cells], P = 0.844). There were no differences in time to recovery of neutrophils and platelets or in the incidences of acute and chronic graft-versus-host disease, overall survival, non-relapse mortality and relapse incidence.ConclusionsDonors >55 years old mobilized fewer CD34⁺ cells and required a greater volume to collect a similar number of CD34⁺ cells. The outcome of HSCT was not influenced by donor age. Donor age should not be a limitation for related allogeneic HSCT.     

The Effects of Mn2+ on the Proliferation, Osteogenic Differentiation and Adipogenic Differentiation of Primary Mouse Bone Marrow Stromal Cells

The effects of Mn²⁺ on the proliferation, osteogenic and adipogenic differentiation of BMSCs were evaluated by employing MTT, ΔΨm, cell cycle, ALP activity, collagen production, ARS and oil red O stain assays. The results indicated that Mn²⁺ decreased the viability at most concentrations for 24 h, but the viability was increased with prolonging incubation time. Mn²⁺ at the concentrations of 1?×?10^-7 and 1?×?10^-6?mol/L decreased ΔΨm in the BMSCs for 48 h. Mn²⁺ induced G2/M phase cell cycle arrest at tested concentrations. On day 7 and 10, the effect of Mn²⁺ on the osteogenic differentiation depended on concentration, but it inhibited osteogenic differentiation at all tested concentrations for 14 d. The effect of Mn²⁺ on the synthesis of collagen of BMSCs depended on concentration for 7 d, but Mn²⁺ inhibited the synthesis of collagen at all tested concentrations for 10 d. On day 14, Mn²⁺ inhibited the formation of mineralized matrix nodules of BMSCs at all tested concentrations, the inhibitory effect turned to be weaker with prolonging incubation time. Mn²⁺ promoted the adipogenic differentiation of BMSCs at all tested concentrations for 10 d, but had no effect with prolonging incubation time. These findings suggested the effects of Mn²⁺ on the proliferation, osteogenic differentiation and adipogenic differentiation of BMSCs are very complicated, concentration and incubation time are key factors for switching the biological effects of Mn²⁺ from damage to protection.     

In vitrouptake and antimycobacterial activity of liposomal usnic acid formulation

The cellular uptake and antimycobacterial activity of usnic acid (UA) and usnic acid-loaded liposomes (UA-LIPOs) were assessed on J774 macrophages. The minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) of UA and UA-LIPO against Mycobacterium tuberculosis were determined. Concentrations required to inhibit 50% of cell proliferation (IC₅₀) were 22.5 (±0.60) and 12.5 (±0.26) μg/ml, for UA and UA-LIPO, respectively. The MICs of UA and UA-LIPO were 6.5 and 5.8 μg/mL, respectively. The MBC of UA-LIPO was twice as low (16 μg/mL) as that of UA (32 μg/mL). An improvement in the intracellular uptake of UA-LIPO was found (21.6 × 10⁴ ± 28.3 × 10² c.p.s), in comparison with UA (9.5 × 10⁴ ± 11.4 × 10² c.p.s). In addition, UA-LIPO remains much longer inside macrophages (30 hours). All data obtained from the encapsulation of usnic acid into liposomes as a drug delivery system (DDS) indicate a strong interaction between UA-liposomes and J774 macrophages, thereby facilitating UA penetration into cells. Considering such a process as ruling the Mycobacterium-transfection by magrophages, we could state that associating UA with this DDS leads to an improvement in its antimycobacterial activity.     

何氏生髓方联合聚乙二醇化重组人粒细胞集落刺激因子防治肺腺癌患者化疗后骨髓抑制的临床效果研究

摘要 目的：探讨何氏生髓方联合聚乙二醇化重组人粒细胞集落刺激因子（PEG-rhG-CSF）防治肺腺癌患者化疗后骨髓抑制的临床效果。方法：选取广州中医药大学东莞医院2021年3月~2022年10月期间收治的100例肺腺癌患者作为研究对象,采用随机数字表法将患者分为对照组（n=50,PEG-rhG-CSF治疗）和观察组（n=50,何氏生髓方联合PEG-rhG-CSF治疗）。观察两组中医证候积分、T淋巴细胞亚群指标、外周血象和中性粒细胞减少发生率变化情况。结果：治疗4个周期后,观察组神疲乏力、胃纳差、食少纳差、少气懒言、自汗盗汗、腰膝酸软症状评分低于对照组（P<0.05）。治疗4个周期后,观察组CD3⁺、CD4⁺、CD4⁺/CD8⁺高于对照组;CD8⁺低于对照组（P<0.05）。治疗4个周期后,观察组白细胞计数（WBC）、血小板计数（PLT）、中性粒细胞计数（NEUT）、血红蛋白（Hb）高于对照组（P<0.05）。观察组中性粒细胞减少发生率低于对照组（P<0.05）。结论：何氏生髓方联合PEG-rhG-CSF防治肺腺癌患者化疗后骨髓抑制,可提高机体免疫力,改善临床症状和外周血象,降低中性粒细胞减少发生率。