癌基因双微体-2在多发性骨髓瘤中的表达及其临床意义

目的探讨癌基因双微体．2（MDM2）在多发性骨髓瘤（MM）中的表达及其临床意义。方法选取2008年7月至2010年6月温州市第二人民医院血液内科MM患者37例,另选5例巨球蛋白血症患者作为疾病对照组,5名骨髓捐献志愿者为健康对照组。应用逆转录聚合酶链式扩增反应和免疫印迹法分别检测各组MDM2mRNA及其蛋白变化。MM患者按国际分期系统将其分为I期11例,Ⅱ期18例,Ⅲ期20例,应用回归分析法观察MDM2mRNA及其蛋白表达与各分期的相关性。结果MDM2mRNA及其蛋白在健康对照组表达水平均很低,在疾病对照组表达水平稍高,在MM组则明显表现为过表达∽均〈0．05）。另外,MDM2mRNA及其蛋白表达与MM分期呈正相关（r值分别为0．782、0．821）。结论MDM2基因在MM患者中呈高水平表达,且与MM的临床分期有密切关系,临床上或可作为判断其预后和检测肿瘤微小残留的一个指标     

自噬体在肺炎链球菌感染过程中的保护作用机制探讨

目的:检测微管相关蛋白轻链-3(LC3)在肺泡Ⅱ型上皮细胞A549上的表达情况,及3-甲基腺嘌呤(3MA)刺激后对其表达的影响,为研究自噬体在抵抗肺炎链球菌感染过程中的保护作用奠定基础.方法:体外培养肺泡Ⅱ型上皮细胞A549,在肺炎链球菌感染A549细胞12h时用倒置显微镜拍照并提取RNA,采用RT-PCR的方法检测LC3 mRNA的表达情况.同时检测对照组、3MA组、Spn组和3MA+ Spn组上清液乳酸脱氢酶(LDH)的OD值.结果:RT-PCR检测LC3 mRNA有明显表达,3MA可以抑制LC3的表达.LDH检测显示Spn组和3MA+ Spn组上清液LDH的OD值数据两两比较差异有统计学意义(P＜0.05).结论:LC3在肺泡Ⅱ型上皮细胞中显著表达,3MA可以抑制细胞的自噬作用.加3MA后,肺炎链球菌感染肺泡Ⅱ型上皮细胞坏死增加,提示自噬体在抵抗肺炎链球菌的感染过程中起一定的保护作用.     

Astragaloside IV attenuates renal tubulointerstitial fibrosis by inhibiting p38 MAPK signaling

Objective To investigate the effect of astragaloside IV (AS-IV) on renal tubulointerstitial fibrosis and its regulation on p38 MAPK signaling. Methods In vivo, UUO model with renal tubulointerstitial injury was constructed. Mice in AS-IV group were orally administrated AS-IV 20 mg•kg-1•d-1 for 7 days after operation, and mice in other groups were administrated the equal volume vehicle. Bilateral kidneys were collected in 7 and 14 days after operation. Transverse kidney slices were stained with Masson trichrome to evaluate the severity of renal tubule injury. In vitro, normal human renal tubular epithelial cells (HK-2) were stimulated with recombinant TGF-β1 (10 ng/ml) and simultaneously treated with different concentrations of AS-IV (0, 50, 100, 200 μg/ml) for 24 h. SB203580 (10 μmol/L) was also ultilized to pre-treat HK-2 cells for 1 h to inhibit phosphorylation of p38 MAPK signaling. The expression of FN, Col IV, and α-SMA were investigated by western blotting and real-time PCR. The expression of p-p38 MAPKs were also observed by Western Blotting. Results Astragaloside IV morphologically ameliorated renal tubulointerstitial fibrosis. The proteins and mRNA expression of FN, Col IV, α-SMA, and TGF-β1 were also increased significantly in UUO kidney tissues (all P＜0.05), which could be reversed by AS-IV administration (all P＜0.05). In vitro, the expression of FN, Col IV, and α-SMA were up-regulated by TGF-β1 after stimulating for 24 h (all P＜0.05), which were decreased by AS-IV. The inhibition effect on FN and α-SMA were similar between AS-IV and MAPK inhibitor SB203580. AS-IV inhibited p-p38 MAPK signals both in vivo and in vitro. Conclusions AS-IV could attenuate renal tubulointerstitial fibrosis induced by UUO and TGF-β1 through reducing FN、Col IV、α-SMA expression in renal tubular cells. The mechanism of AS-IV protective effect might be associated with inhibition of p38 MAPK phosphorylation.     

Trpm5 expression in the olfactory epithelium

The Ca²⁺-activated monovalent cation channel Trpm5 is a key element in chemotransduction of taste receptor cells of the tongue, but the extent to which Trpm5 channels are expressed in olfactory sensory neurons (OSNs) of the main olfactory epithelium (MOE) of adult mice as part of a specific pheromonal detection system is debated. Here, we used a novel Trpm5-IRES-Cre knockin strain to drive Cre recombinase expression, employed previously validated Trpm5 antibodies, performed in situ hybridization experiments to localize Trpm5 RNA, and searched extensively for Trpm5 splice variants in genetically-labeled, Trpm5-expressing MOE cells. In contrast to previous reports, we find no evidence for the existence in adult mouse OSNs of the classical Trpm5 channel known from taste cells. We show that Trpm5-expressing adult OSNs express a novel Trpm5 splice variant, Trpm5-9, that is unlikely to form a functional cation channel by itself. We also demonstrate that Trpm5 is transiently expressed in a subpopulation of mature OSNs in the embryonic olfactory epithelium, indicating that Trpm5 channels could play a specific role in utero during a narrow developmental time window. Ca²⁺ imaging with GCaMP3 under the control of the Trpm5-IRES-Cre allele using a newly developed MOE wholemount preparation of the adult olfactory epithelium reveals that Trpm5-GCaMP3 OSNs comprise a heterogeneous group of sensory neurons many of which can detect general odorants. Together, these studies are essential for understanding the role of transient receptor potential channels in mammalian olfaction.     

Transnasal surgery for prolactin-secreting pituitary adenomas in childhood and adolescence

BACKGROUND

Prolactin (PRL)-secreting pituitary adenomas (prolactinomas) are rare in childhood and adolescence; there are only a few published series of patients who were surgically treated. We discuss the gender-dependent differences, and the surgical indications and results for 14 patients with prolactinomas under 18 years of age at surgery.

METHODS

We reviewed the data for 14 patients (10 girls and 4 boys) treated surgically for prolactinomas between 1980 and 1996. Surgery was chosen because of intolerance and/or resistance to dopamine agonist (DA) in 9 patients, the patient’s decision in 3, rhinorrhea in 1, and pituitary apoplexy in 1 during DA therapy. All patients underwent direct transnasal explorations. Since 1988, intraoperative PRL measurements were performed. The follow-up period was at least 6 years.

RESULTS

Nine of the 10 girls had primary or secondary amenorrhea, and 3 of the 4 boys had visual field defects. The boys and girls were the same age at the onset of symptoms and had the same preoperative duration of symptoms. Boys had larger, more invasive tumors and higher preoperative and postoperative plasma PRL levels than girls. Two boys demonstrated significant extrasellar extension at the time of diagnosis. Two patients required drilling of the incompletely pneumatized sphenoid sinus to reach the sella. Radical tumor resection was achieved in seven girls and in none of the four boys. In 12 of the 14 patients, normal pituitary function was preserved by transnasal surgery. There was no severe surgical morbidity or mortality. Endocrinological remission by surgery alone was achieved for seven girls, and was achieved in 4 of the 5 patients (80%) with microadenomas. In patients with preoperative mean PRL levels of less than 200 μg/L, the surgical cure rate was 75%.

CONCLUSIONS

Prolactinomas in childhood and adolescence differ in their biology in boys and girls. Transnasal surgery is as safe in childhood patients as it is in adult patients, and it may be an effective alternative to long-term medical therapy for selected patients with prolactinomas. Successful surgical outcomes were achieved in patients with microadenomas and preoperative serum PRL levels of less than 200 μg/L.     

Absence of close linkage between Hereditary Spherocytosis (SPH) and 24 genetic marker systems including HLA and GM

A total of 250 individuals belonging to 19 different families, identified through established propositi were simultaneously screened for hereditary spherocytosis (SPH), using stringent criteria, and 27 well-known polymorphic genetic marker systems.
The segregation analysis indicated that the pattern of inheritance of SPH in these families, being autosomal and dominant, had a 100 % penetrance. A statistical analysis, using the LIPED computer program of Ott (1974), revealed the absence of close linkage between SPH and AB0, ACP1, ADA, AK1, C3, D1A2, ESD, FY, CC, GL01, GM, GPT1, HLA, HPA, JK, K, KM, MNS, P, PGD, PGM1, PGP, PI , and RH.
Since an earlier study by other investigators had convincingly suggested a linkage between SPH and GM , we subjected the data to further analysis and found no significant heterogeneity in our recombination values of linkage between SPH and GM , or any of the other informative loci.     

Alteration of Human Leukocyte Chernotaxis, Chemiluminescence, and HMP Shunt Activity Caused By Mechanical Trauma

The effect of graded shear stress exposure (0 to 800 dynes/cm² at 35°C) on human leukocyte function was examined in vitro, using a concentric cylinder viscometer. Boyden Chamber chemotaxis was significantly reduced at stress levels above 150 dynes/cm², and was over a factor of three lower after exposure to trauma levels of 450 dynes/cm² Hexose monophosphate (HMP) shunt activity following particle ingestion was also significantly lower after exposure to shear stresses of 150 dynes/cm² or more for ten minutes. PMN chemiluminescence was reduced following stress trauma of 450 dynes/cm² using whole blood, and after 150 dynes/cm² using leukocyte suspensions. All the functional alterations reported appear to be bulk shear stress effects and not due to surface interaction, as changing the surface-to-volume ratio in the viscometer by over a factor of three did not change the damage caused at a given stress level. These functional alterations caused by mechanical trauma may be important in hemodialysis, cardiopulmonary bypass, venovenous perfu-sion, and in any blood-contacting artificial organ. The stress levels required for damage are of the same order as those required for platelet functional alteration, and much lower than those required for erythrocyte hemolysis for the same exposure times.     

Isolation of Fibrinogen–Fibrin Related Antigen from Human Plasma by Immunoaffinity Chromatography: its Characterization in Normal Subjects and in Defibrinating Patients with Abruptio Placentae and Disseminated Cancer

Summary. Highly purified, fibrinogen–fibrin related antigen (FR-antigen) was isolated with good recovery from 1.0–2.0 ml of human plasma, by immunoaffinity chromatography with antibody specific for fibrinogen and fibrin, and plasmin degradation products X, Y, D and D-D dimer. In FR-antigen from defibrinating patients there was evidence for thrombin activity alone (mainly disseminated cancer) or both plasmin and thrombin (mainly abruptio placentae). Thus, the molar ratio of N-terminal Gly – Tyr in the FR-antigen of 18 of 20 patients strongly suggested thrombin activity (95th percentile). In addition, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on unreduced samples frequently showed bands similar in mol wt to fragments X, Y and D, and in the reduced samples A α and Bβ chain degradation, both indicating plasmin activity. N-terminal β chain Ala was elevated in the antigen of four of 20 patients, also suggesting plasmin activity (99th percentile). Combined thrombin, plasmin and factor-XIII activity, as shown by bands in similar position to D-D and γ chain dimers, was commonly associated with high levels of serum FR-antigen (> 10 mg/dl). In some defibrinating patients, especially those with disseminated cancer, heterogeneity of unreduced FR-antigen and Aα chain degradation, both indicators of mild plasmin-like activity which are commonly seen in normals, were absent.