Effect of urogastrone on intestinal regeneration is dose-dependent

Urogastrone (UG) increases the rate of intestinal regeneration in intestinal defects patched with adjacent serosal surfaces by increasing the rate of epithelial cell migration and proliferation. It also inhibits contraction of the patched intestinal defect. The purpose of this study was to determine the effect of the dose of UG on these processes. Twenty male New Zealand white rabbits (1.9-2.9 kg) had 2 x 5 cm ileal defects patched with adjacent caecal serosal surface. Group I (n = 6) served as the control group. Group II (n = 5), Group III (n = 5) and Group IV (n = 4) received UG 0.15, 1.5 and 4.5 micrograms/kg/h i.v. via mini-osmotic pumps. Seven days after patching, both epithelialization of the patched defect and neomucosal surface area were significantly increased by UG and the increases were dose-dependent. Contraction was not inhibited by the lowest dose of UG but was diminished by 1.5 micrograms/kg/h. Proliferative activity in both neomucosa and adjacent normal mucosa were increased in UG-treated animals with the greatest increase in rabbits receiving 1.5 and 4.5 micrograms/kg/h UG. The effect of UG on both epithelialization and contraction of patched intestinal defects is dose-dependent. Since the lowest dose of urogastrone increased epithelialization without increasing proliferative activity, stimulated cell migration appears to be the earliest effect of UG.     

Effect of Urogastrone On Intestinal Regeneration Is Dose-Dependent

Urogastrone (UG) increases the rate of intestinal regeneration in intestinal defects patched with adjacent serosal surfaces by increasing the rate of epithelial cell migration and proliferation. It also inhibits contraction of the patched intestinal defect. the purpose of this study was to determine the effect of the dose of UG on these processes. Twenty male New Zealand white rabbits (1.9–2.9 kg) had 2 × 5 cm ileal defects patched with adjacent caecal serosal surface. Group I (n=6) served as the control group. Group II (n=5), Group III (n=5) and Group IV (n=4) received UG 0.15, 1.5 and 4.5 μg/kg/h i.v. via mini-osmotic pumps. Seven days after patching, both epithelialization of the patched defect and neomucosal surface area were significantly increased by UG and the increases were dose-dependent. Contraction was not inhibited by the lowest dose of UG but was diminished by 1.5 μg/kg/h. Proliferative activity in both neomucosa and adjacent normal mucosa were increased in UG-treated animals with the greatest increase in rabbits receiving 1.5 and 4.5 μg/kg/h UG. the effect of UG on both epithelialization and contraction of patched intestinal defects is dose-dependent. Since the lowest dose of urogastrone increased epithelialization without increasing proliferative activity, stimulated cell migration appears to be the earliest effect of UG.     

Tissue engineered esophagus by copper——small intestinal submucosa graft for esophageal repair in a canine model

Acellular porcine small intestinal submucosa(SIS)has been used for esophagoplasty with success in a canine model.However,it did not lead to complete epithelialization.For better reconstruction,a cellular component is required.Moreover,promotion of angiogenesis with copper has been widely recognized by basic research as well as clinical studies.In this study,we have evaluated the feasibility and effectiveness of combined Cu and SIS(SIS-Cu patch)for the esophageal repair using a canine model.Eighteen male beagle dogs were subjected to surgical resection to produce cervical esophageal defects(5 cm in length,180°in range).SIS with Cu(5 or 25μmol L 1copper)or without Cu was patched on the esophageal defects.Barium esophagram and histology exam were carried out to evaluate the effectiveness of the therapy.As shown,the SIS-Cu graft promoted re-epithelialization,re-vascularization and muscular regeneration.SIS-Cu patch is more effective than SIS alone for esophageal repair,and the SIS+25μmol L 1Cu group demonstrated additional advantages over the SIS+5μmol L 1Cu.     

Effect of the duration of infusion of urogastrone on intestinal regeneration in rabbits

Urogastrone (UG) increases the rate of mucosal regeneration on patched intestinal defects. Our aim was to determine the effect of the duration of UG administration on regeneration of serosa patched ileal defects in rabbits. Group I (n = 18) were controls. Group II (n = 15), Group III (n = 10) and Group IV (n = 5) received UG 1.5 micrograms/kg/h subcutaneously for 7 days, 14 days or 21 days respectively. Animals were sacrificed at 7 day intervals up to 21 days after patching. Neomucosal growth was significantly greater in the animals receiving UG and was greatest in Group IV. Group II and III animals had less contraction of the patched defect and greater neomucosal surface area than Group I animals at each interval but had a lesser effect than animals receiving UG continuously. Crypt cell production rate was significantly greater in UG treated animals at 7 and 14 days but fell to control levels at 21 days. Prolonging the duration of UG infusion increased the quantity of neomucosa produced by intestinal regeneration. However, UG stimulation of mucosal cell migration and proliferation occurred transiently within 14 days after patching.     

DHEA administration has limited effect onintestinal Ca absorption in ovariectomized rats

[Purpose]

The effect of dehydroepiandrosterone (DHEA) administration on intestinal Calcium (Ca) absorption in estrogen deficiency state has not been studied yet. We examined the bone mineral content (BMC) of lumbar spine and Ca balance such as intestinal Ca absorption and Ca accumulation in ovariectomized (OVX) rats after 8 weeks of DHEA administration.

[Methods]

Seventeen female Sprague-Dawley rats, 6 weeks old, were randomized into two groups: OVX control rats (OC, n = 8) and OVX rats with DHEA treatment (OD, n = 9). DHEA was administered to the OD group intraperitoneally at 20 mg DHEA/kg body weight for 8 weeks while the OC group was treated with vehicle only.

[Results]

The BMC normalized by body weight of the lumbar spine (trabecular-abundant region) in the OD group was found to be significantly higher compared to that in the OC group. The femoral wet weight normalized by body weight in the OD group was significantly higher compared to that in the OC group. The intestinal Ca absorption, rate of intestinal Ca absorption, Ca accumulation, and rate of Ca accumulation decreased from the 4th and 5th of the experimental diet period to the end of the experimental period, but interaction of time and group was not observed. In both periods, all parameters did not differ between the groups.

[Conclusion]

The present study confirmed the positive effect of DHEA on trabecular bone mass in ovariectomized rats. On the other hand, DHEA administration might have limited the impact on intestinal Ca absorption in estrogen deficiency state.     

创伤性脑损伤对大鼠海马骨架蛋白及学习记忆功能影响

创伤性脑损伤（traumatic brain injury,TBI）是极为常见的外伤性疾病,致死率和致残率很高。存活者伴随的空间认知功能障碍,给患者家庭和社会造成了极大的负担。目前,对TBI造成的空间记忆障碍缺乏系统研究。脑损伤后海马组织与记忆有关的分子以及组成神经元骨架的分子如何变化研究甚少。本研究采用Wistar大鼠为研究对象,并随机将其分为假手术（sham）组和创伤性脑损伤（TBI）组。TBI组再按致伤后时间长短分为6 h、12 h、24 h、72 h、15 d五个亚组。TBI组应用PinPointTM颅脑撞击器撞击而致伤,sham组不撞击。采用Morris水迷宫评价实验动物空间记忆能力;干湿重法测定脑含水量,评估脑水肿与海马水通道蛋白4（aquaporin-4,AQP-4）的相关性;海马神经元特异性核蛋白（neuron specific nuclear protein,NeuN）标记和免疫荧光检测评估TBI致大鼠神经元丢失情况;通过Western印迹检测TBI致海马骨架相关蛋白质和记忆相关蛋白质含量变化。本研究证实,与sham组相比,TBI组大鼠潜伏期明显增加［（61.98±12.82) s vs.(28.32±8.52) s,n=5,P<0.01,day 15］,探索时间明显缩短［（36.98±0.37) s vs. (73.68±5.09) s,n=5,P<0.01,day15］,表明脑创伤损害了动物的空间参考记忆能力和空间工作记忆能力。与sham组相比,TBI组大鼠海马AQP-4在蛋白质水平上的表达和脑含水量持续升高,15 d恢复正常;在12 h［（3.78±0.74),(83.78±0.35)%］和72 h［（3.49±0.85),(82.28±0.63)%］均形成两个波峰,n=5,P均<0.01,表明继发性脑损伤与持续脑水肿和海马AQP-4在蛋白质上的高表达有关。与sham组相比,NeuN标记和免疫荧光检测发现,TBI后24 h 致大鼠海马神经元丢失严重［（198.2±8.002) vs.(297.2±6.866) cells/mm2, n=5,P<0.01］,表明TBI动物的海马功能受损。与sham相比,TBI组海马神经元树突标志物微管结合蛋白2（microtubule associated proein 2,MAP2）和突触前终末特异性标记物突触素（synaptophysin,SYN）在蛋白质水平均伤后逐步降低（n=5,P均<0.01）,72 h［（0.55±0.05) vs.(1.27±0.08), (0.52±0.14) vs.(1.06±0.16), n=5,P均<0.01］降低最明显;TBI组形成神经元纤维缠结主要成分的过度磷酸化tau（ser404）,伤后逐步升高,72 h［（1.25±0.11)vs. (0.33±0.07), n=5,P<0.01］升高最明显。 MAP2、SYN和过度磷酸化的tau（ser404）检测指标的改变,表明脑损伤致神经元受损,神经元生长和损伤修复能力减弱,最终导致神经元骨架破环,TBI损害了动物的海马空间记忆能力。与sham组相比,TBI组大鼠海马环磷酸腺苷反应元件结合蛋白（cAMP response element binding protein,CREB）和磷酸化CREB ser133(phosphorylated CREB Ser133, pCREB Ser133)含量降低明显（n=5,P均<0.05）,表明脑损伤动物海马的存储记忆能力减弱;TBI组大鼠海马一般调控阻遏蛋白激酶2(general control nonderepressible 2 kinase,GCN2)蛋白质升高明显（n=5,P均<0.05）,表明脑损伤动物海马将新信息转化成长期记忆能力下降。本研究提示,创伤性脑损伤可使大鼠海马神经元骨架破坏,进而导致在学习记忆过程中起重要作用的分子蛋白质下调,抑制记忆储存的蛋白质（GCN2）上调,促使学习记忆功能障碍。     

创伤性脑损伤对大鼠海马骨架蛋白及学习记忆功能影响

创伤性脑损伤（traumatic brain injury,TBI）是极为常见的外伤性疾病,致死率和致残率很高。存活者伴随的空间认知功能障碍,给患者家庭和社会造成了极大的负担。目前,对TBI造成的空间记忆障碍缺乏系统研究。脑损伤后海马组织与记忆有关的分子以及组成神经元骨架的分子如何变化研究甚少。本研究采用Wistar大鼠为研究对象,并随机将其分为假手术（sham）组和创伤性脑损伤（TBI）组。TBI组再按致伤后时间长短分为6 h、12 h、24 h、72 h、15 d五个亚组。TBI组应用PinPointTM颅脑撞击器撞击而致伤,sham组不撞击。采用Morris水迷宫评价实验动物空间记忆能力;干湿重法测定脑含水量,评估脑水肿与海马水通道蛋白4（aquaporin-4,AQP-4）的相关性;海马神经元特异性核蛋白（neuron specific nuclear protein,NeuN）标记和免疫荧光检测评估TBI致大鼠神经元丢失情况;通过Western印迹检测TBI致海马骨架相关蛋白质和记忆相关蛋白质含量变化。本研究证实,与sham组相比,TBI组大鼠潜伏期明显增加［（61.98±12.82) s vs.(28.32±8.52) s,n=5,P<0.01,day 15］,探索时间明显缩短［（36.98±0.37) s vs. (73.68±5.09) s,n=5,P<0.01,day15］,表明脑创伤损害了动物的空间参考记忆能力和空间工作记忆能力。与sham组相比,TBI组大鼠海马AQP-4在蛋白质水平上的表达和脑含水量持续升高,15 d恢复正常;在12 h［（3.78±0.74),(83.78±0.35)%］和72 h［（3.49±0.85),(82.28±0.63)%］均形成两个波峰,n=5,P均<0.01,表明继发性脑损伤与持续脑水肿和海马AQP-4在蛋白质上的高表达有关。与sham组相比,NeuN标记和免疫荧光检测发现,TBI后24 h 致大鼠海马神经元丢失严重［（198.2±8.002) vs.(297.2±6.866) cells/mm2, n=5,P<0.01］,表明TBI动物的海马功能受损。与sham相比,TBI组海马神经元树突标志物微管结合蛋白2（microtubule associated proein 2,MAP2）和突触前终末特异性标记物突触素（synaptophysin,SYN）在蛋白质水平均伤后逐步降低（n=5,P均<0.01）,72 h［（0.55±0.05) vs.(1.27±0.08), (0.52±0.14) vs.(1.06±0.16), n=5,P均<0.01］降低最明显;TBI组形成神经元纤维缠结主要成分的过度磷酸化tau（ser404）,伤后逐步升高,72 h［（1.25±0.11)vs. (0.33±0.07), n=5,P<0.01］升高最明显。 MAP2、SYN和过度磷酸化的tau（ser404）检测指标的改变,表明脑损伤致神经元受损,神经元生长和损伤修复能力减弱,最终导致神经元骨架破环,TBI损害了动物的海马空间记忆能力。与sham组相比,TBI组大鼠海马环磷酸腺苷反应元件结合蛋白（cAMP response element binding protein,CREB）和磷酸化CREB ser133(phosphorylated CREB Ser133, pCREB Ser133)含量降低明显（n=5,P均<0.05）,表明脑损伤动物海马的存储记忆能力减弱;TBI组大鼠海马一般调控阻遏蛋白激酶2(general control nonderepressible 2 kinase,GCN2)蛋白质升高明显（n=5,P均<0.05）,表明脑损伤动物海马将新信息转化成长期记忆能力下降。本研究提示,创伤性脑损伤可使大鼠海马神经元骨架破坏,进而导致在学习记忆过程中起重要作用的分子蛋白质下调,抑制记忆储存的蛋白质（GCN2）上调,促使学习记忆功能障碍。     

miR-491-5p靶向调控SHOC2对食管鳞癌细胞增殖、迁移及侵袭的影响

目的探讨miR-491-5p对食管鳞癌细胞增殖、迁移及侵袭的影响及作用机制。 方法培养永生化食管上皮细胞株HET-1A和人食管癌细胞株EC109,EC9706,KYSE510,qRT-PCR检测细胞中miR-491-5p和富含亮氨酸重复蛋白SHOC2 （SHOC2） mRNA水平。EC109细胞分为空白对照组、miR- 491-5p组、miR-NC组、miR-491-5p+pcDNA-SHOC2组和miR-491-5p+pcDNA组,MTT检测细胞增殖,Transwell检测细胞迁移和侵袭,Western Blot法检测CyclinD1、Vimentin、E-cadherin以及MAPK/ERK信号通路相关蛋白水平。双荧光素酶报告基因实验验证miR- 491- 5p与SHOC2之间调控关系。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,两两比较采用SNK-q检验。 结果食管鳞癌细胞EC109、EC9706和KYSE510中miR-491-5p表达水平低于HET-1A细胞（0.32±0.06、0.62±0.10、0.61±0.08比1.00±0.08）,差异具有统计学意义（F = 106.340,P < 0.001）;SHOC2 mRNA表达水平高于HET- 1A细胞（2.85±0.16、1.73±0.10、1.45±0.06比1.02±0.09）,差异具有统计学意义（F = 464.949,P < 0.001）。miR-491-5p组EC109细胞培养72 h后的OD值、细胞迁移数、侵袭数及CyclinD1、Vimentin、p-MEK和p-ERK蛋白水平均低于miR-NC组（0.70±0.06比1.42±0.08,65.01±10.36比150.01±12.48,70.03±10.26比140.02±11.85,0.30±0.03比0.93±0.16,0.41±0.05比0.86±0.08,0.32±0.06比0.95±0.11,0.40±0.06比0.92±0.13）,差异具有统计学意义（F = 236.565、159.440、120.706、101.071、98.619、130.766、77.046,P均< 0.001）,E-cadherin蛋白水平高于miR-NC组（0.89±0.13比0.48±0.08）,差异具有统计学意义（F = 816.432,P < 0.001）。miR-491-5p在EC109细胞中负调控SHOC2表达,SHOC2过表达逆转了miR-491-5p过表达对EC109细胞增殖、迁移和侵袭及MAPK/ERK信号通路的影响。 结论miR-491-5p可抑制食管鳞癌细胞的增殖、迁移和侵袭,其作用机制可能与下调SHOC2表达抑制MAPK/ERK信号通路活性有关。     

miR-214通过调控Notch1信号对口咽鳞状细胞癌细胞株增殖、侵袭及凋亡的作用机制研究

目的研究miR-214通过调控Notch1信号对口咽鳞状细胞癌（OSCC）细胞株增殖、侵袭及凋亡的作用机制。 方法在ATCC细胞库购买OSCC细胞株,分为3组：抑制组（IN组）、模拟组（SI组）及对照组（CO组）。通过Western blot法、qRT-PCR法、TUNEL法、侵袭实验及CCK-8法分析3组癌细胞中Notch1的蛋白、mRNA表达水平、细胞凋亡、迁移及增殖能力的差异性,三组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。 结果IN、SI、CO组细胞内miR-214mRNA表达量分别为1.15±0.26,3.34±0.89,2.08±0.67,SI组分别与IN组、CO组比较,差异具有统计学意义（t = 5.281,P = 0.002;t = 2.529,P = 0.045）;SI组、IN组的Notch1-UTR表达分别为0.42±0.11、0.86±0.09,差异具有统计学意义（t = 5.362,P = 0.006）;IN、SI、CO组的OSCC细胞凋亡率分别为（0.78±0.10）﹪、（0.32±0.06）﹪、（0.56±0.12）﹪,IN组分别与CO组、SI组比较,差异具有统计学意义（t = 3.149,P = 0.020;t = 8.823,P < 0.001）;IN、SI、CO组癌细胞的增殖率分别为（0.65±0.05）﹪、（1.26±0.06）﹪、（1.89±0.04）﹪,IN组与SI组、IN组与CO组比较,差异具有统计学意义（t = 11.056,P < 0.001;t = 27.394,P < 0.001）,CO组与SI组比较,差异具有统计学意义（t = 12.365,P < 0.001）;IN、SI、CO组癌细胞侵袭率分别为（0.13±0.02）﹪、（0.89±0.13）﹪、（0.48±0.11）﹪,SI组与CO组、SI组与IN组比较,差异具有统计学意义（t = 4.176,P = 0.006;t = 10.147,P < 0.001）;IN、SI、CO组的Notch1蛋白含量分别为：2.38±1.34、0.94±0.23、1.76±0.67,SI组与CO组、SI组与IN组比较,差异具有统计学意义（t = 2.588,P = 0.041;t = 2.368,P = 0.056）;IN、SI、CO组的Notch1 mRNA表达量分别为4.26±1.06、1.45±0.38、2.46±0.87,IN组与CO组、IN组与SI组比较,差异具有统计学意义（t = 2.935,P = 0.026;t = 5.583,P = 0.001）。 结论miR-214可能与OSCC细胞株增殖呈负相关,在细胞中miR-214含量升高,可能抑制Notch1信号通路表达,从而促进癌细胞增长、侵袭,抑制癌细胞凋亡。     

硫酸酯酶2型基因体内表达促进化疗药物诱发的小鼠骨髓抑制恢复作用研究

采用半定量RT-PCR和重组基因体内表达法观察了硫酸酯酶2基因（Sulfatase 2,Sulf2）在5-氟脲嘧啶（5-Fluorouracil, 5-Fu）诱发的小鼠骨髓抑制和再生过程中作用。结果表明：Sulf2在小鼠骨髓抑制和再生过程中呈现先上升,后下降的动态表达;电转pcDNA3.1-Sulf2基因实验组外周血白细胞数和血小板数在5-Fu注射后第7天分别为(1216.7±457.9)/μl和(8.1±5.4)万/μl,明显低于对照组［分别为(1691.7±228.9)/μl和(14.7±2.1)万/μl］,实验组单条腿骨髓细胞总数在第7天为(94.2±21.1)万,显著低于对照组（173±59.9）万,但在第11天为(585±337.9)万,又显著地高于对照组（255±65.3）万,实验组第7天10000个骨髓细胞总集落形成数为(9±8.4),显著低于对照组（39±12.2),统计均有显著性差异（p<0.05）。这些结果提示Sulf2可能对5 Fu诱发的小鼠骨髓抑制后的再生具有促进作用。     

骨髓间充质干细胞动员内源性干细胞修复放射性肠黏膜损伤的实验研究

目的探索骨髓间充质干细胞（MSCs）对放射性肠炎（RE）肠黏膜修复的途径。 方法体外分离、培养大鼠骨髓MSCs。将RE模型的大鼠采用随机数字表法分为治疗组[经尾静脉注射干细胞悬液1 mL（细胞浓度为1×10⁶个/mL）]和对照组（注射等量生理盐水,每日1次,连续3 d）,每组10只。每日观察两组大鼠的活动量、进食和进水量、体质量变化等。1周后处死大鼠获取小肠标本,HE染色观察肠黏膜的修复情况,电镜观察上皮细胞的超微结构,免疫组化染色观察小肠隐窝Bmi-1阳性干细胞增殖情况,采用Image-Pro Plus 6.0软件对Bmi-1阳性细胞数量进行分析。组间差异的比较采用t检验。 结果成功分离得到大鼠骨髓MSCs,流式细胞仪鉴定：CD29、CD90、CD34、CD45阳性细胞比例分别为98.6﹪、99.6﹪、0.56﹪、0.89﹪。与对照组相比,治疗组大鼠移植1周后,体质量增加（190.30 g ± 13.23 g比235.00 g±14.30 g）;大鼠小肠黏膜上皮得到修复,绒毛高度增高（627.50 μm ± 40.55 μm比984.33 μm ± 61.80 μm）;上皮细胞超微结构较完整、清晰,隐窝Bmi-1阳性干细胞增殖数量增多[（60.67±9.63）个/mm²比（87.33 ±5.47）个/mm²],差异具有统计学意义（P < 0.05）。 结论骨髓MSCs能促进RE模型的大鼠肠黏膜的修复,这一作用可能是通过促进小肠隐窝干细胞的增殖发挥。     

Functional characteristics of Peyer's patch lymphoid cells. IV. Effect of antigen feeding on the frequency of antigen-specific B cells.

The frequency of B cells in Peyer's patches from normal BDF(1) and sheep red blood cell (SRBC)-fed BDF(1) mice that could respond to antigenic determinants on SRBC and trinitrophenyl (TNP) was determined using an in vitro system of limiting dilution analysis. In normal mice, one B cell in 1.9 x 10(4) Peyer's patch cells could be induced to an anti-SRBC response and one B cell in 3.6 x 10(4) Peyer's patch cells could be induced to an anti-TNP response. The frequency of B cells capable of responding to SRBC in normal mice was similar in Peyer's patches and spleen. However, after feeding mice SRBC for 3 weeks, there was a 6-fold reduction in the frequency of B cells in Peyer's patches capable of responding to SRBC but no change in the frequency of B cells capable of responding to TNP. The average clone size of Peyer's patch B cells responding to SRBC was similar in normal and SRBC-fed mice. Although antigen-feeding does not stimulate Peyer's patch B cells in situ to humoral antibody synthesis, antigen-feeding can markedly alter the reactivity of the antigen-sensitive cell population in Peyer's patches. We previously demonstrated that T cells in Peyer's patches could be specifically carrier primed for helper function by SRBC feeding. We have now demonstrated that antigen-feeding reduced significantly the frequency of B cells in Peyer's patches capable of responding to the fed antigen. Peyer's patches appear to serve an important function as a sampling site for intestinal antigens.     

自体骨髓干细胞移植联合奥扎格雷对糖尿病足的临床疗效探究

目的探讨自体骨髓干细胞移植联合奥扎格雷对糖尿病足（DF）的临床疗效。 方法选取2017年7月至2018年8月期间于重庆市大渡口区人民医院收治的99例DF患者为研究对象,随机分为奥扎格雷组、移植组与联合组3组各33例。全部患者入院后均给予DF的常规治疗,奥扎格雷组、移植组、联合组分别给予奥扎格雷、自体骨髓干细胞移植、自体骨髓干细胞移植联合奥扎格雷,比较干预前、干预后12周的临床症状体征、DF严重程度、生存质量的变化,以及干预后12周的新生侧支血管的分级情况。定性资料采用χ²检验或Wilcox秩和检验,定量资料组内干预前后比较采用配对t检验,多组比较先采用方差分析,然后采用Tukey检验进行两两比较。采用Pearson相关系数探讨截肢组患者生活质量评分的相关因素。 结果3组患者干预前的基线资料、疼痛、冷感、间歇性跛行的评分、踝肱指数（ABI）、Wagner分级、糖尿病特异性生活质量量表（DSQL）各维度评分与总分比较,差异无统计学意义（P > 0.05）,具有可比性。组内比较,3组干预后的疼痛、冷感、间歇性跛行的评分、DSQL生理功能、心理（精神）因素、社会关系等维度评分与总分均低于干预前,Wagner分级、ABI优于干预前,差异有统计学意义（P < 0.05）。组间比较,联合组干预后的疼痛、冷感、间歇性跛行的评分、DSQL生理功能、心理（精神）因素等维度评分与总分分别为0.83±0.36、0.83±0.31、1.36±0.63、8.9±3.2、7.5±2.5、23.7±9.2,均低于奥扎格雷组的1.13±0.39、1.26±0.59、1.89±0.73、12.5±5.2、10.1±3.1、31.7±8.8及移植组的1.08±0.33、1.11±0.55、1.72±0.60、10.9±3.6、9.3±3.3、28.8± 7.6,差异有统计学意义（F = 5.001、5.598、3.953、2.230、9.610,P均< 0.05）。联合组干预后的ABI为0.55±0.21,高于奥扎格雷组的0.43±0.20及移植组的0.42±0.16,差异有统计学意义（F = 4.051,P < 0.05）。联合组干预后的社会关系维度评分、Wagner分级为1级的比例分别为5.0±2.1、81.8%,高于奥扎格雷组的6.3±2.3、54.5%,差异有统计学意义（F = 3.953,χ²= 6.983,P均 < 0.05）。观察组干预后的新生侧支血管分级优于奥扎格雷组,差异有统计学意义（P < 0.05）。 结论自体骨髓干细胞移植联合奥扎格雷能缓解DF患者的临床症状,促进溃疡愈合,促进移植术后的血管新生,提高生存质量,值得临床推广应用。     

Effect of the duration of infusion of urogastrone* on intestinal regeneration in rabbits

Abstract Urogastrone (UG) increases the rate of mucosal regeneration on patched intestinal defects. Our aim was to determine the effect of the duration of UG administration on regeneration of serosa patched ileal defects in rabbits. Group I (n= 18) were controls. Group II (n= 15), Group III (n= 10) and Group IV (n= 5) received UG 1.5 μ/kg/h subcutaneously for 7 days, 14 days or 21 days respectively. Animals were sacrificed at 7 day intervals up to 21 days after patching. Neomucosal growth was significantly greater in the animals receiving UG and was greatest in Group IV. Group II and III animals had less contraction of the patched defect and greater neomucosal surface area than Group I animals at each interval but had a lesser effect than animals receiving UG continuously. Crypt cell production rate was significantly greater in UG treated animals at 7 and 14 days but fell to control levels at 21 days. Prolonging the duration of UG infusion increased the quantity of neomucosa produced by intestinal regeneration. However, UG stimulation of mucosal cell migration and proliferation occurred transiently within 14 days after patching.     

血毒清联合杂合肾脏替代治疗对重症脓毒症的疗效、血流动力学和预后干预作用的研究

目的探讨血毒清联合杂合肾脏替代治疗（HRRT）重症脓毒症的效果及对血流动力学、预后的影响。 方法选取2017年3月至2019年8月十堰市太和医院重症脓毒症患者135例,在常规治疗基础上,按简单随机化法分为血毒清组、HHRT组和血毒清联合HRRT组（联合组）,每组45例,治疗5 d。治疗前和治疗5 d后序贯器官衰竭评分（SOFA）、急性生理学与慢性健康状况评分系统Ⅱ评分（APACHEⅡ）、炎症因子指标[白细胞介素-10（IL-10）、IL-6、降钙素原（PCT）、C反应蛋白（CRP）]水平、血流动力学指标[全身血管阻力指数（SVRI）、血管外肺水指数（EVLWI）、胸腔内血容积指数（ITBVI）、心指数（CI）]及重症监护室（ICU）住院时间比较采用配对t检验,多组间差异采用方差分析,组间两两比较采用LSD-t检验,3组28 d病死率采用c²检验。 结果治疗5 d后,与联合组比较,HHRT组和血毒清组SOFA评分[（4.29±1.17）分比（6.15±1.39）分、（7.03±1.52）分]、APACHEⅡ评分[（11.68±2.12）分比（14.26±3.04）分、（15.17±2.85）分]、血清IL-10 [（36.61±6.50）μg/L比（42.75±7.42） μg/ L、（45.06±8.37） μg / L]、IL-6 [（130.26±41.04） pg/ mL比（169.84±46.75） pg/ mL、（178.36±50.91） pg/ mL]、PCT [（0.87±0.29） ng/mL比（1.96±0.47） ng/mL、（2.24±0.53）ng/mL]、CRP [（34.93±9.71） mg/ L比（51.67±10.32）mg/L、（56.79±11.82）mg/L] 、EVLWI [（4.12±1.38）mL/kg比（5.38±1.69） mL/ kg、（6.04±1.85）mL/kg]降低,而SVRI [（2079.54±124.75） dyn·s·cm^{- 5}·m²比（1865.37±109.38） dyn·s·cm^-5·m²、（1796.28±131.75）dyn·s·cm^-5·m²]、ITBVI [（1189.40±92.36） mL/ m²比（986.97±75.32）mL/m²、（902.37±68.64） mL/ m²]、CI [（4.26±0.65） L·min^-1·m^-2比（3.83±0.58）L·min^-1·m^-2、（3.67±0.53）L·min^-1·m^-2]均升高,差异均有统计学意义（P < 0.05）。与联合组比较,HHRT组和血毒清组ICU住院时间[（14.82±4.40）d比（17.20±3.47）d、（18.46±4.25）d]缩短（P < 0.05）。 结论血毒清联合HRRT治疗重症脓毒症,对减少炎性因子水平,调节血流动力学具有一定作用,能够缩短ICU住院时间。     

硫化氢通过调控SIRT1抑制阿霉素诱导的H9c2细胞损伤

目的探讨硫化氢（H₂S）对阿霉素（DOX）诱导的H9c2细胞损伤的影响及其作用机制。 方法H₂S对DOX心肌毒性保护作用的实验分组为：对照组（Control组）,5?μmol/?L DOX处理组（A组）,5?μmol/L DOX和400?μmol/L NaHS共同处理组（B组）,400?μmol/L NaHS单独处理组（C组）,5?μmol/L DOX、400?μmol/L NaHS和15?μmol/L Sirtinol共同处理组（D组）,15?μmol/L Sirtinol单独处理组（E组）。SIRT1是否参与H₂S抗DOX心肌毒性作用机制的实验分组为：对照组（Control组）,5?μmol/L DOX处理组（F组）,5?μmol/L DOX和400?μmol/L NaHS共同处理组（G组）,5?μmol/L DOX、400?μmol/L NaHS和15?μmol/L Sirtinol共同处理组（H组）,15?μmol/L Sirtinol单独处理组（I组）。使用MTT法检测细胞活力;Elisa法检测细胞MDA以及SOD水平;DCFH-?DA荧光探针法检测ROS水平;采用Western Blot法检测SIRT1蛋白表达。使用单因素方差分析法进行统计学分析。 结果NaHS预处理可抑制DOX导致的H9c2细胞活力下降：Control组,A组、B组、C组细胞活力分别为100﹪、（54.58±1.58）﹪、（85.05±4.31）﹪、（100.22±4.46）﹪ （F = 134.9,P < 0.001）。NaHS预处理可减弱DOX引起的H9c2细胞ROS、MDA水平的增加以及SOD水平的降低：Control组的ROS、MDA和SOD水平分别是100﹪、（34.18±1.56） μmol/g、（53.69±1.44） U/?mg;A组的ROS、MDA和SOD水平分别是（174.90±12.65）﹪、（72.65±2.66） μmol/g、（31.80±2.05） U/?mg;B组的ROS、MDA和SOD水平分别是（126.08±6.25）﹪、（44.59±1.92） μmol/g、（48.06±1.56） U/mg;C组的ROS、MDA和SOD水平分别是（91.86±1.66）﹪、（32.93±1.56）?μmol/?g、（55.93±1.58）?U/?mg （F?= 83.26,P < 0.001;F = 271.4,P < 0.001;F = 127.0,P < 0.001）。F组（6、12、24?h）H9c2细胞SIRT1蛋白表达水平分别是（0.45±0.03）、（0.27±0.02）、（0.25±0.03）,较Control组（1.00±0.00）降低（F = 611.1,P < 0.001）。本研究还发现,NaHS预处理H9c2细胞能阻止DOX引起的SIRT1蛋白表达下调：Control组、F组、G组、H组的SIRT1蛋白表达水平分别是（1.00±0.00）、（0.31±0.03）、（0.60±0.04）、（1.09±0.09）（F = 123.4,P?2S对DOX诱导的H9c2细胞活力降低的抑制作用：Control组,F组、G组、H组、I组细胞活力分别为100﹪、（54.58±1.58）﹪、（85.37±3.62）﹪、（71.11±2.11）﹪、（97.53±1.45）﹪ （F = 238.2,P < 0.001）。Sirtinol预处理可明显逆转H₂S对DOX导致的H9c2细胞ROS和MDA含量增加及SOD水平降低的抑制作用：Control组的ROS、MDA和SOD水平分别是100﹪、（35.84±2.22）μmol/?g、（53.03±3.16） U/mg;F组的ROS、MDA和SOD水平分别是（184.6±11.33）﹪、（74.78±5.30）μmol/g、（29.26±0.85）U/mg;G组的ROS、MDA和SOD水平分别是（126.5±7.57）﹪、（41.95±3.43）μmol/g、（52.61±2.26）U/mg;H组的ROS、MDA和SOD水平分别是（174.7±5.50）﹪、（67.69±1.52） μmol/g、（35.33±1.95） U/mg,I组的ROS、MDA和SOD水平分别是（98.03±2.86）﹪、（37.66±2.49）μmol/g、51.14 U/mg（F = 112.0,P < 0.001;F = 93.73,P < 0.001;F = 84.92,P < 0.001）。 结论H₂S通过调控SIRT1抑制DOX诱导的H9c2细胞损伤。     

AngⅡ通过激活Notch1/Sox2通路对肝星状细胞LX2的活化和增殖的作用

目的探讨AngⅡ通过靶向调控Notch1/Sox2肝星状细胞LX2细胞的增殖。 方法通过CCK8检测AngⅡ对肝星状细胞增殖能力的影响,通过羟脯氨酸酸法检测AngⅡ对肝星状细胞胶原合成能力的影响,并通过脂质体转染siRNA-Notch1构建Notch1低表达细胞模型,通过CCK8检测敲低Notch1对AngⅡ诱导的肝星状细胞LX2增殖的影响,通过Western Blot检测敲低Notch1对AngⅡ诱导的肝星状细胞LX2蛋白表达的影响。所有的检测结果都进行生物学重复。采用方差分析和t检验进行统计学分析。 结果CCK8结果显示,AngⅡ（5、10、20、40、80 nmol/L）预处理A450值分别为0.67±0.06、0.88±0.07、0.98±0.07、1.08±0.07、1.23±0.07,较对照组0.57±0.05上升,差异具有统计学意义（F = 45.76,P < 0.01）,羟脯氨酸检查结果显示,AngⅡ（10、20、40 nmol/L）预处理组羟脯氨酸浓度分别为（2.60±0.20）、（3.47±0.25）、（4.17±0.21）mg/L,羟脯氨酸浓度较对照组（1.90±0.10）mg/L上升,差异具有统计学意义（F = 75.18,P < 0.01）。Western Blot结果显示,AngⅡ10、20、40 nmol/L组Notch1蛋白表达水平分别为0.20±0.02、0.54±0.04、0.82±0.03,与正常对照组0.11±0.02发生升高,差异具有统计学意义（F = 400.50,P < 0.01）。Notch1干扰后,CCK8结果显示,siRNA-Notch1+AngⅡ组（10、20、40 nmol/L）A450值分别为0.53±0.06、0.83±0.03、1.03±0.03,与siRNA-NC+ AngⅡ对照组0.97±0.06,1.43±0.06,1.73±0.06比较发生降低（P < 0.01）。进一步Western Blot结果显示,Notch1敲低组（AngⅡ+ siRNA-Notch1）Notch1、HES1和Sox2蛋白表达水平分别为1.47±0.12、0.77±0.06和0.50±0.10,分别与AngⅡ对照组2.83±0.15、2.20±0.10和1.17±0.06比较,差异具有统计学意义（P < 0.01）。 结论AngⅡ通过激活Notch1/Sox2信号促进肝星状细胞LX2增殖。     

Combined transmyocardial revascularization and cell-based angiogenic gene therapy increases transplanted cell survival

We hypothesized that pretreatment of an infarcted heart by mechanical transmyocardial revascularization (TMR) before transplantation of bone marrow cells (BMCs) or BMC-expressing angiogenic growth factors would increase transplanted BMC survival and enhance myocardial repair. Female Lewis rats underwent coronary ligation 3 wk before creation of 10 needle TMR channels (3 groups) or no TMR (3 groups), followed by transplantation of 3 x 10(6) male donor BMCs, BMC transfected with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and insulin-like growth factor-1 (IGF-1) (BMC + VBI), or medium alone. At 1, 3, and 7 days, we evaluated transplanted cell survival, vascular densities, and left ventricular (LV) function (N = 4 per group x 6 groups x 3 time points). At 3 days, vascular densities in the scar were increased by TMR + BMC + VBI and by BMC + VBI (P < 0.05), and at 7 days, vascular densities were greatest in rats receiving TMR + BMC + VBI (P < 0.05). Transplanted cell survival at 3 and 7 days was increased by TMR and by BMC + VBI. Combined therapy with TMR + BMC + VBI resulted in the greatest cell survival at 3 days (P < 0.05) versus BMC. After 7 days, LV ejection fraction (LVEF) was lowest in rats receiving neither BMC nor TMR and greatest in rats receiving TMR + BMC + VBI (P = 0.004). We concluded that mechanical pretreatment of infarcted myocardium by TMR enhances the effect of subsequent cell-based gene therapy on transplanted cell survival, angiogenesis, and LV function. Scar pretreatment with TMR combined with cell-based multigene therapy may maximize myocardial repair.     

Evaluation of the cholesterol influence in type II collagen-induced arthritis in DBA/1J mice: an autoradiographic study

In order to verify the cholesterol influence in RA severity in DBA/1J mice, we quantified the cholesterol present in the knee joints of normal (N) and with collagen II induced arthritis (CIA). Forty male DBA/1J mice, were divided in normal (n=20) and CIA group (n=20). Mice in CIA group were injected with 100 μg of collagen II emulsified in Freund's complete adjuvant. Sixteen DBA/1J (8 N and 8 CIA) received an injection of 2.96 × 10⁶ Bq of 3H-cholesterol and were anesthetized and sacrificed. Semi-fine sections were covered with LM-1 emulsion, exposed for six weeks and developed. Collagen induced edema, erythema and dysfunction of knee joints in CIA group. Radioactive cholesterol was located more on the synovial membrane, where we found the greatest density of silver grains, significantly ( P <0.0001) higher in group CIA vs. controls (61±2.3 × 18±0.7). We conclude that the cholesterol deposits on the synovial membrane is related to CIA severity.     

利拉鲁肽联合二甲双胍对肥胖合并2型糖尿病患者糖脂代谢、胰岛β细胞功能及体脂的影响探究

目的探讨利拉鲁肽联合二甲双胍对肥胖合并2型糖尿病患者糖脂代谢、胰岛β细胞功能及体脂的影响。 方法选择成都市郫都区第二人民医院2017年1月至2018年12月收治的肥胖合并2型糖尿病患者141例作为研究对象,按照随机数字表法分为利拉鲁肽组47例、二甲双胍组47例与联合组47例。利拉鲁肽组采用利拉鲁肽治疗,二甲双胍组采用二甲双胍治疗,联合组采用利拉鲁肽联合二甲双胍治疗。三组疗程均为3个月。比较三组治疗前后糖代谢、脂代谢、胰岛β细胞功能和体重指数（BMI）变化,及不良反应,治疗前后比较采用配对t检验,三组间比较采用F检验,两组间比较采用LSD-t检验。 结果三组治疗后FPG、HbA1c和2hPG水平较治疗前降低,差异具有统计学意义（P均< 0.05）;联合组治疗后FPG（6.57±0.39）?mmol/L、HbA1c（7.03±0.42）%和2hPG（8.78±0.45）mmol/L低于利拉鲁肽组（7.03±0.32）mmol/ L、（7.68±0.35）%、（9.56±0.65）mmol/L（t = 6.251、8.151、6.764,P均?< 0.05）和二甲双胍组（7.06±0.39）mmol/L、（7.76±0.46）%、（9.70±0.81）?mmol/L,差异具有统计学意义（t = 6.091、8.034、6.807,P均< 0.05）。三组治疗后HDL-C水平较治疗前升高,而TC、LDL-C和TG水平较治疗前降低,差异具有统计学意义（P均< 0.05）;联合组治疗后HDL-C （1.56±0.13） mmol/ L高于利拉鲁肽组（1.29±0.14） mmol/ L（t = 9.689,P < 0.05）和二甲双胍组（1.32±0.15） mmol/ L,差异具有统计学意义（t = 8.289,P < 0.05）;而联合组TC（4.35±0.38） mmol/L、LDL-C（2.79±0.21）mmol/L和TG（2.15±0.26） mmol/ L低于利拉鲁肽组（5.18±0.43）mmol/L、（3.19±0.15）mmol/L、（2.65±0.17） mmol/L（t = 9.916、10.626、11.035,P < 0.05）和二甲双胍组（5.15±0.34）mmol/L、（3.23±0.25）mmol/ L、（2.68±0.23） mmol/ L,差异具有统计学意义（t = 10.756、9.239、10.467,P均< 0.05）。三组治疗后HOMA-β较治疗前升高,差异具有统计学意义（P < 0.05）;联合组治疗后HOMA-β（4.87±0.28）高于利拉鲁肽组（4.15±0.36）和二甲双胍组（4.08±0.41）,差异具有统计学意义（t = 10.823,10.909,P均< 0.05）。三组治疗后BMI较治疗前降低,差异具有统计学意义（P < 0.05）;联合组治疗后BMI （28.08±0.37）kg/m²低于利拉鲁肽组（29.73±0.49）kg/m²和二甲双胍组（29.61±0.43）kg/m²,差异具有统计学意义（t = 18.423,18.490,P均< 0.05）。 结论利拉鲁肽联合二甲双胍对肥胖合并2型糖尿病患者效果良好,可改善患者糖脂代谢和胰岛β细胞功能,降低体质指数,值得临床借鉴。