小鼠肝星状细胞的分离、培养及鉴定方法

目的：探索BALB／c小鼠肝星状细胞（HSCs）分离、纯化的可行方案．并评价依该法分离所得HSCs的生物学特性。方法：经肝门静脉先用不含钙镁离子的Hank液充分灌洗,再以浓度为1mg／mL的Ⅳ型胶原酶灌注BALB／c小鼠肝脏。取肝后．完整分离后碾碎,37℃水浴振荡30min,Percoll连续密度梯度（60％）离心法分离、纯化HSCs．苔盼蓝染色检测HSCs活性,结蛋白免疫细胞化学鉴定HSCs,光镜观察体外培养HSCs的形态学变化。结果：纯化后每只小鼠HSCs收获量约为（5．5±0．4）×10^5个．HSCs纯度〉90％．HSCs细胞活率〉90％。结论：本实验建立的的分离纯化方案可获得高纯度高活率的小鼠HSCs。     

Adaptation of Tibial Structure and Strength to Axial Compression Depends on Loading History in Both C57BL/6 and BALB/c Mice

Tibial compression can increase murine bone mass. However, loading protocols and mouse strains differ between studies, which may contribute to conflicting results. We hypothesized that bone accrual is influenced more by loading history than by mouse strain or animal handling. The right tibiae of 4-month-old C57BL/6 and BALB/c mice were subjected to axial compression (10 N, 3 days/week, 6 weeks). Left tibiae served as contralateral controls to calculate relative changes: (loaded ? control)/control. The WashU protocol applied 60 cycles/day, at 2 Hz, with a 10-s rest-insertion between cycles; the Cornell/HSS protocol applied 1,200 cycles/day, at 6.7 Hz, with a 0.1-s rest-insertion. Because sham loading, sedation, and transportation did not affect tibial morphology, unhandled mice served as age-matched controls (AC). Both loading protocols were anabolic for cortical bone, but Cornell/HSS loading elicited a more rapid response that was greater than WashU loading by 13 %. By 6 weeks, cortical bone volume of each loading group was greater than of AC (average + 16 %) and not different from each other. Ultimate displacement and energy to fracture were greater in tibiae loaded by either protocol, and ultimate force was greater with Cornell/HSS loading. At 6 weeks, independent of mouse strain, the WashU protocol produced minimal trabecular bone and the trabecular bone volume fraction of Cornell/HSS tibiae was greater than that of AC by 65 % and that of WashU by 44 %. We concluded that tibial adaptation to loading was more influenced by waveform than mouse strain or animal handling and therefore may have targeted similar osteogenic mechanisms in C57BL/6 and BALB/c mice.     

去势对雄性BALB/c小鼠早期血清性激素及蛋白表达等影响

目的 探讨雄性小鼠去势术后早期内环境变化特点,为合理有效建立前列腺良恶性动物模型在去势与非去势的选择方面提供参考. 方法 选用16只BALB/c雄性小鼠,随机平均分为去势组和对照组.去势组采用经阴囊途径双睾丸切除术,对照组则作同一切口假手术.第21天所有小鼠采血处死,分别检测血清睾酮(T)、雌二醇(E2)、血管内皮生长因子(VEGF)和SELDI蛋白分析. 结果 与对照组相比,去势组血清T接近成倍升高(730/400),E2反而成倍降低(112.53/256.45),T/E2较对照组升高约4倍(6.49/1.56);VEGF变化不明显(P＞0.05);去势组在7个蛋白位点(3.4、5.3、7.1、11.6、11.8、15.2和15.9 KDa)有高表达,在2个位点(6.0和6.2 KDa)为低表达. 结论 去势与否不仅对小鼠机体性激素、VEGF和蛋白表达等有影响,而且会导致在去势与否基础上所建立的前列腺良恶性模型特征不同及其研究结果差异.     

Inbred Strain‐Specific Response to Biglycan Deficiency in the Cortical Bone of C57BL6/129 and C3H/He Mice

Inbred strain‐specific differences in mice exist in bone cross‐sectional geometry, mechanical properties, and indices of bone formation. Inbred strain‐specific responses to external stimuli also exist, but the role of background strain in response to genetic deletion is not fully understood. Biglycan (bgn) deficiency impacts bone through negative regulation of osteoblasts, resulting in extracellular matrix alterations and decreased mechanical properties. Because osteoblasts from C3H/He (C3H) mice are inherently more active versus osteoblasts from other inbred strains, and the bones of C3H mice are less responsive to other insults, it was hypothesized that C3H mice would be relatively more resistant to changes associated with bgn deficiency compared with C57BL6/129 (B6;129) mice. Changes in mRNA expression, tissue composition, mineral density, bone formation rate, cross‐sectional geometry, and mechanical properties were studied at 8 and 11 wk of age in the tibias of male wildtype and bgn‐deficient mice bred on B6;129 and C3H background strains. Bgn deficiency altered collagen cross‐linking and gene expression and the amount and composition of mineral in vivo. In bgn's absence, changes in collagen were independent of mouse strain. Bgn‐deficiency increased the amount of mineral in both strains, but changes in mineral composition, cross‐sectional geometry, and mechanical properties were dependent on genetic background. Bgn deficiency influenced the amount and composition of bone in mice from both strains at 8 wk, but C3H mice were better able to maintain properties close to wildtype (WT) levels. By 11 wk, most properties from C3H knockout (KO) bones were equal to or greater than WT levels, whereas phenotypic differences persisted in B6;129 KO mice. This is the first study into mouse strain‐specific changes in a small leucine‐rich proteoglycan gene disruption model in properties across the bone hierarchy and is also one of the first to relate these changes to mechanical competence. This study supports the importance of genetic factors in determining the response to a gene deletion and defines biglycan's importance to collagen and mineral composition in vivo.     

Relationship of Glycated Albumin to Blood Glucose and HbA1c Values and to Retinopathy,Nephropathy, and Cardiovascular Outcomes in the DCCT/EDIC Study

The association of chronic glycemia, measured by HbA_1c, with long-term complications of type 1 diabetes has been well established in the Diabetes Control and Complications Trial (DCCT) and other studies. The role of intermediate-term and acute glycemia and of glucose variability on microvascular and cardiovascular disease (CVD) is less clear. In order to examine the interrelationships among long-term, intermediate-term, and acute measures of glucose and its daily variability, we compared HbA_1c, glycated albumin (GA), and seven-point glucose profile concentrations measured longitudinally in a case-cohort subpopulation of the DCCT. HbA_1c and GA were closely correlated with each other and with the mean blood glucose (MBG) calculated from the seven-point profile. The associations of glucose variability and postprandial concentrations with HbA_1c and GA were relatively weak and were further attenuated when MBG was included in multivariate models. In the case-cohort analyses, HbA_1c and GA had similar associations with retinopathy and nephropathy, which were strengthened when both measures were considered together. Only HbA_1c was significantly associated with CVD. The demonstrated interrelationships among different measures of glycemia will need to be considered in future analyses of their roles in the development of long-term complications of type 1 diabetes.     

Orbital and/or zygomatic removal in an approach to lesions near the cranial base. Surgical technic, anatomic study and analysis of a series of 24 cases

The authors describe the technique of removal of the orbital rim, the zygomatic arch and the combination of both, i.e., the orbito-zygomatic arch. Then they give the summary of an anatomical study in human cadavers (13 approaches), measuring the field view angle (F.V.A.) of various intra-cranial targets in the vicinity of the skull base after orbital and/or zygomatic removal associated to fronto-temporal craniotomy. Thanks to orbital and/or zygomatic removal, F.V.A. was increased on average, by 75% in the sub-frontal approach, 46% in the pterional approach, and 86% in the sub-temporal approach. Finally the authors illustrate the usefulness of these combined approaches in a series of 24 cases of various difficult lesions located in the vicinity of the skull base, operated on during the last 3 years.     

Effect of Isoflurane and Other Potent Inhaled Anesthetics on Minimum Alveolar Concentration, Learning, and the Righting Reflex in Mice Engineered to Express α1 γ-Aminobutyric Acid Type A Receptors Unresponsive to Isoflurane

Background: Enhancement of the function of [gamma]-aminobutyric acid type A receptors containing the [alpha]1 subunit may underlie a portion of inhaled anesthetic action. To test this, the authors created gene knock-in mice harboring mutations that render the receptors insensitive to isoflurane while preserving sensitivity to halothane.

Methods: The authors recorded miniature inhibitory synaptic currents in hippocampal neurons from hippocampal slices from knock-in and wild-type mice. They also determined the minimum alveolar concentration (MAC), and the concentration at which 50% of animals lost their righting reflexes and which suppressed pavlovian fear conditioning to tone and context in both genotypes.

Results: Miniature inhibitory postsynaptic currents decayed more rapidly in interneurons and CA1 pyramidal cells from the knock-in mice compared with wild-type animals. Isoflurane (0.5-1 MAC) prolonged the decay phase of miniature inhibitory postsynaptic currents in neurons of the wild-type mice, but this effect was significantly reduced in neurons from knock-in mice. Halothane (1 MAC) slowed the decay of miniature inhibitory postsynaptic current in both genotypes. The homozygous knock-in mice were more resistant than wild-type controls to loss of righting reflexes induced by isoflurane and enflurane, but not to halothane. The MAC for isoflurane, desflurane, and halothane did not differ between knock-in and wild-type mice. The knock-in mice and wild-type mice did not differ in their sensitivity to isoflurane for fear conditioning.     

An International Urogynecological Association (IUGA) / International Continence Society (ICS) joint terminology and classification of the complications related directly to the insertion of prostheses (meshes, implants, tapes) & grafts in female pelvic floor surgery

Introduction and hypothesis

A terminology and standardized classification has yet to be developed for those complications arising directly from the insertion of synthetic (prostheses) and biological (grafts) materials in female pelvic floor surgery.

Methods

This report on the above terminology and classification combines the input of members of the Standardization and Terminology Committees of two International Organizations, the International Urogynecological Association (IUGA) and the International Continence Society (ICS) and a Joint IUGA/ICS Working Group on Complications Terminology, assisted at intervals by many expert external referees. An extensive process of 11 rounds of internal and external review took place with exhaustive examination of each aspect of the terminology and classification. Decision-making was by collective opinion (consensus).

Results

A terminology and classification of complications related directly to the insertion of prostheses and grafts in female pelvic floor surgery has been developed, with the classification based on category (C), time (T) and site (S) classes and divisions, that should encompass all conceivable scenarios for describing insertion complications and healing abnormalities. The CTS code for each complication, involving three (or four) letters and three numerals, is likely to be very suitable for any surgical audit or registry, particularly one that is procedure-specific. Users of the classification have been assisted by case examples, colour charts and online aids (www.icsoffice.org/complication).

Conclusions

A consensus-based terminology and classification report for prosthess and grafts complications in female pelvic floor surgery has been produced, aimed at being a significant aid to clinical practice and research.     

NeuroD/BETA2 gene variability and diabetes: no associations to late-onset type 2 diabetes but an A45 allele may represent a susceptibility marker for type 1 diabetes among Danes. Danish Study Group of Diabetes in Childhood, and the Danish IDDM Epidemiology and Genetics Group

Mutations in the NeuroD/BETA2 gene have been shown to associate with type 2 diabetes. In the present study, we examined mutations in the NeuroD/BETA2 gene for association with either type 1 or 2 diabetes. Three variants were identified in patients with type 2 diabetes: Ala45Thr (allelic frequency 0.36, 95% CI 0.31-0.41), Pro197His (0.01), and Ser259Ser (0.01). Ala45Thr and Pro197His were not associated with type 2 diabetes, but the transmission disequilibrium test showed unequal transmission of the A45 allele to offspring with type 1 diabetes (chi2 = 5.90, P < 0.02, odds ratio 1.55, 95% CI 0.91-2.63). This association could not be explained by linkage disequilibrium between the Ala45 allele and IDDM7 (D2S152), which is also located on chromosome 2q32. When tested in vitro, the biological activity of Thr45 (117+/-36% vs. Ala45) and His197 (90+/-28% vs. Pro197) on the regulation of the human insulin gene promoter appeared normal. In conclusion, mutations in the NeuroD/BETA2 gene are not a common cause of late-onset type 2 diabetes among Danes. However, in the type 1 diabetic Danish population, the Ala45Thr variant of NeuroD/BETA2 may represent a susceptibility marker independent of IDDM7 on chromosome 2q32.