蒺藜皂苷对环磷酰胺致小鼠生精细胞凋亡的保护作用

目的研究蒺藜皂苷对环磷酰胺(cyclophosphamide,CP)所致小鼠生精细胞凋亡的保护作用,探讨蒺藜皂苷对生殖功能障碍小鼠的保护机制。方法将8周龄的雄性昆明种小鼠随机分为3组,对照组采取腹腔注射生理盐水,模型组和治疗组采取连续5d腹腔注射CP 80mg/(kg·d),腹腔注射同时将对照组和模型组小鼠连续灌胃生理盐水,治疗组小鼠连续灌胃蒺藜皂苷200mg/(kg·d)35d。比较各组小鼠体质比及睾丸重量;检测附睾精子密度、活精子百分率、精子活力和精子畸形率;HE染色观察睾丸生精上皮的发育;TUNEL法检测生精细胞的凋亡;Western blot检测睾丸组织中凋亡相关蛋白Caspase-8表达水平的变化。结果与模型组比较,蒺藜皂苷显著提高了生精功能损伤小鼠的精子密度、活精子百分比、精子活力,降低了精子畸形率,促进了生精小管上皮精原细胞及精子细胞的发育,上调了睾丸组织中Caspase-8的表达水平,使生精细胞的凋亡减少。结论蒺藜皂苷能显著修复CP所致的小鼠睾丸损伤,其修复机制可能通过上调Caspase-8的表达,拮抗环磷酰胺引起的生精功能损伤,减少生精细胞的凋亡。     

大肠埃希菌感染诱导小鼠生精细胞凋亡的研究

目的探讨大肠埃希菌感染对雄性小鼠生精细胞凋亡以及对凋亡调控基因Fas/FasL的影响,进一步阐明大肠埃希菌感染致男性不育的分子机理。方法用大肠埃希菌直接注入成年雄性小鼠膀胱内模拟逆行感染的途径,感染后第7d处死小鼠,取睾丸组织分别作HE染色和用TUNEL法观察生精细胞凋亡及睾丸的病理变化;免疫组化法检测Fas/FasL在睾丸组织中的表达,同时检测精子数量、精子活动率和精子畸形率。结果大肠埃希菌感染小鼠后生精细胞的凋亡指数及Fas和FasL表达水平分别为(18.50±1.50)%、(48.10±4.19)%和(34.60±4.18)%,对照组分别为(7.40±1.69)%、(23.50±2.66)%和(19.10±4.30)%,差异均有统计学意义(P均<0.05)。感染组精子数量、精子活动率和精子畸形率分别为(6.00±1.41)×106/ml、(59.80±5.29)%和(14.78±2.03)%,对照组分别为(12.00±4.12)×106/ml、(88.90±5.37)%和(5.88±1.10)%,感染组和正常对照组比较差异均有统计学意义(P均<0.01)。感染组小鼠睾丸组织出现明显的病理变化。结论雄性小鼠感染大肠埃希菌后,可导致生精细胞凋亡增加及上调Fas和FasL蛋白的表达水平;Fas和FasL在小鼠睾丸组织的高表达诱导生精细胞的凋亡,最终使小鼠精子形成减少,活动率下降,畸形率增加。大肠埃希菌感染造成生精细胞凋亡可能是导致男性不育的原因之一。     

无精子症睾丸组织中survivin、Caspase-3的表达及意义

目的观察survivin、Caspase-3在不同病理类型无精子症睾丸组织中的表达变化及相关性。方法用免疫组化方法检测44例无精子症患者睾丸组织中的survivin、Caspase-3。结果唯支持细胞综合征、睾丸生精阻滞、生精功能低下和睾丸生精未见异常的睾丸组织中survivin阳性表达率分别为18．18％、81．82％、90．91％、100．00％（P均〈0．05）,Caspase-3阳性表达率分别为45．45％、36．36％、36．36％、18．18％（P均〉0．05）。survivin与Caspase-3的表达呈负相关（r=-0．636,P〈0．05）。结论在人睾丸组织中有survivin与Caspase-3的表达,survivin与Caspase-3在生精过程中可能发挥协同作用。     

转录因子Tbx18及Wt1在小鼠心脏发育过程中的时空表达

目的:原位观察转录因子Tbx18及Wt1在胚胎心脏中的表达,及心肌细胞本身是否表达Tbx18及Wt1。方法:收集不同发育阶段小鼠胚胎(E)心脏,冰冻切片后,取不同区域的组织进行免疫荧光染色和DAPI染核,检测Tbx18、Wt1及Nkx2.5的表达。结果:小鼠前体心外膜及不同发育阶段的心外膜可明显表达Tbx18及Wt1蛋白,但未检测到心脏转录因子Nkx2.5的表达。从E10.5~至少E14.5d,Tbx18蛋白明显表达于不同区域的心脏组织中。除E14.5 d少许表达Tbx18的细胞不表达Nkx2.5外,这些表达Tbx18的细胞还同时表达Nkx2.5。从E12.5~至少14.5 d,Wt1表达于不同区域心脏组织中,但这些表达Wt1的细胞都不表达NKx2.5。结论:从E10.5~至少14.5 d,Tbx18表达于小鼠心肌细胞中;从E12.5d~至少E14.5d,Wt1表达于小鼠心脏组织中,但不表达于心肌细胞中。     

植物乳杆菌对IL-10基因敲除结肠炎小鼠黏附分子的影响

目的: 观察植物乳杆菌( Lactobacillus plantarum,LP)灌胃对IL-10基因敲除小鼠肠道炎症和淋巴细胞归巢的影响.方法: 取IL-10基因敲除(knockout,KO)小鼠和未作基因敲除的背景鼠分为4组: 对照组(野生组WT)、加植物乳杆菌组(WT+LP)、IL-10基因敲除模型组(KO)、模型加植物乳杆菌组(KO+LP).4 wk开始对照组和KO组每日予PBS灌胃,WT+LP和KO+LP组予溶于PBS的LP灌胃,持续4-8 wk结束.实验结束后取各组小鼠结肠行炎症评分和电镜亚显微结构观察,并用RT-PCR和Western blot检测归巢相关分子MAdCAM-1、ICAM-1、α4β7及CD3的表达.结果: 8 w k 后K O小鼠1 0 0%发生肠道炎症,且其CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较WT组均明显增高(mRNA: t = 39.42,8.83,25.53,45.78,均P＜0.01;CD3、ICAM-1、MAdCAM-1蛋白: t = 19.04,29.57,12.29,均P＜0.01).予以益生菌LP灌胃后,KO+LP组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较KO组均明显降低(mRNA: t = 20.34;4.95;14.21;22.31,均P＜0.01;CD3、ICAM-1、MAdCAM-1蛋白: t = 6.82,14.10,7.03,均P＜0.01);WT+L P组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA较WT组均明显降低( t = 9.33,10.55,7.75,6.69,均P＜0.01),而WT+LP组小鼠CD3及黏附分子ICAM-1和MAdCAM-1的蛋白表达水平无明显降低.结论: 植物乳杆菌能下调黏附分子在IL-10基因敲除结肠炎小鼠中的高表达,这可能是其减轻炎症状态,缓解炎症性肠病的重要机制之一.     

维生素E和氟化物对雄性小鼠生殖功能的联合作用

目的　探讨维生素E(VE)和氟化物对雄性小鼠生殖功能的联合作用。方法　选择健康成年雄性小鼠 ,按体重随机分成 5组 :空白对照组、VE对照组、VE拮抗组、染氟 1组和染氟 2组 ,每组 10只。染氟方式为小鼠饮用含氟化钠 2 0 0mg/L(VE拮抗组、染氟 1组 )或 3 0 0mg/L(染氟 2组 )的去离子水 ,VE采用灌胃方式每天给予 2 0 0mg/kg(VE对照组、VE拮抗组 )。每天 1次观察动物并每周称量小鼠体重 1次 ,饲养 5周后 ,检测各组小鼠的血氟水平 ,附睾及睾丸的脏器系数 ,附睾尾精子数、精子活动度及精子畸形率。结果　染氟 2组、染氟 1组及VE拮抗组大部分小鼠于受试后 3～ 5周均出现不同程度氟斑牙 ,各组差异无显著性 (P >0 .0 5 ) ;染氟 2组血氟水平明显高于其他各组 (P <0 .0 1) ,染氟 1组、VE拮抗组的血氟水平也明显高于空白对照组、VE对照组 (P <0 .0 1) ;各组小鼠体重、睾丸及附睾脏器系数、附睾尾精子数及精子畸形率差异无显著性 (P >0 .0 5 ) ;而染氟 2组、染氟 1组、VE拮抗组的活精率及精子活动度明显低于空白对照组和VE对照组 (P <0 .0 5或P <0 .0 1)。结论　饮用含氟化钠 2 0 0mg/L或 3 0 0mg/L的去离子水的雄性小鼠活精率及精子活动度降低 ,染氟同时给予VE ,未见明显改善作用     

细辛、杜仲及其合剂对亚急性衰老小鼠睾丸及血清睾酮影响的实验研究

目的研究细辛、杜仲及其合剂对亚急性衰老小鼠睾丸、精子及血清睾酮的影响。方法选昆明系雄性小鼠,用D-半乳糖制备衰老模型。应用光镜、电镜技术检测细辛、杜仲及其合剂治疗前后衰老小鼠睾丸及精子的形态学改变;放射免疫技术检测血清睾酮的变化。结果衰老小鼠睾丸重量减轻,生精细胞缺如,精子密度及活动率下降,血清睾酮含量明显降低(模型组与青年组比较:P<0.01);用药后,睾丸重量增加,生精小管增粗,生精细胞增多,生精过程活跃,精子密度及活动率明显提高,血清睾酮含量增加(P<0.05)。合剂组的作用优于杜仲组和细辛组(P<0.05)。结论细辛,杜仲及其合剂可改善衰老小鼠的生精功能,明显抑制衰老小鼠血清睾酮含量的下降,具有一定延缓衰老的作用。     

弓形虫急性感染小鼠睾丸的病理学及发病机理的研究揪

目的观察弓形虫急性感染小鼠睾丸的病理学变化及探讨其发病机理.方法将弓形虫急性感染和正常对照NIH小鼠睾丸做印片及切片,观察生精细胞的病理变化及弓形虫侵入细胞的情况;应用免疫组化S-P法进行弓形虫抗原及bcl-2、c-myc、P53、bax的检测.结果弓形虫急性感染组小鼠睾丸印片中见生精细胞胞质及核内弓形虫速殖子;睾丸切片病理变化为生精停滞,精原细胞胞质空泡性变.免疫组化染色显示弓形虫;bcl-2及c-myc在感染组和正常组间的表达无统计学意义(P＞0.05);P53及bax在两组间均未表达.结论弓形虫急性感染小鼠睾丸生精停滞等病理变化可能是弓形虫及其分泌的毒素直接损害、干扰生精细胞生成及分裂发生障碍和死亡的结果,与凋亡基因无明显关系.     

金属硫蛋白3对快速老化痴呆小鼠海马胶原纤维酸性蛋白表达的影响

目的 研究金属硫蛋白3(MT3)对快速老化痴呆模型小鼠(SAMP8)海马结构的胶原纤维酸性蛋白(GFAP)表达的影响.方法 给小鼠不同浓度MT3及盐水腹腔注射28 d,处死小鼠后应用免疫组化和免疫印迹方法检测海马结构GFAP的表达.结果 8月龄的SAMP8小鼠海马结构内可见大量浓染的GFAP阳性星形胶质细胞,GFAP含量最多.随着给予MT3浓度的增加,SAMP8小鼠海马结构内的GFAP阳性细胞减少,GFAP含量减少.结论 SAMP8小鼠海马结构星形胶质细胞增生,GFAP表达增多.MT3有减低GFAP表达的作用,随着浓度增加作用增强.     

弓形虫急性感染小鼠睾丸的病理学及发病机理的研究

目的　观察弓形虫急性感染小鼠睾丸的病理学变化及探讨其发病机理。方法　将弓形虫急性感染和正常对照NIH小鼠睾丸做印片及切片 ,观察生精细胞的病理变化及弓形虫侵入细胞的情况 ;应用免疫组化S -P法进行弓形虫抗原及bcl 2、c myc、P53、bax的检测。结果　弓形虫急性感染组小鼠睾丸印片中见生精细胞胞质及核内弓形虫速殖子 ;睾丸切片病理变化为生精停滞 ,精原细胞胞质空泡性变。免疫组化染色显示弓形虫 ;bcl 2及c myc在感染组和正常组间的表达无统计学意义 (P >0 .0 5 ) ;P53及bax在两组间均未表达。结论　弓形虫急性感染小鼠睾丸生精停滞等病理变化可能是弓形虫及其分泌的毒素直接损害、干扰生精细胞生成及分裂发生障碍和死亡的结果 ,与凋亡基因无明显关系     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.