Improvement of the sentinel lymph node detection rate of cervical sentinel lymph node biopsy using real‐time fluorescence navigation with indocyanine green in head and neck skin cancer

The standard technique using lymphoscintigraphy, blue dye and a gamma probe has established a reliable method for sentinel node biopsy for skin cancer. However, the detection rate of cervical sentinel lymph nodes (SLN) is generally lower than that of inguinal or axillary SLN because of the complexity of lymphatic drainage in the head and neck region and the “shine‐through” phenomenon. Recently, indocyanine green fluorescence imaging has been reported as a new method to detect SLN. We hypothesized that fluorescence navigation with indocyanine green in combination with the standard technique would improve the detection rate of cervical sentinel nodes. We performed cervical sentinel node biopsies using the standard technique in 20 basins of 18 patients (group A) and using fluorescence navigation in combination with the standard technique in 12 basins of 16 patients (group B). The mean number of sentinel nodes was two per basin (range, 1–4) in group A and three per basin (range, 1–5) in group B. The detection rate of sentinel nodes was 83% (29/35) in group A and 95% (36/38) in group B. The false‐negative rate was 6% (1/18 patients) in group A and 0% in group B. Fluorescence navigation with indocyanine green may improve the cervical sentinel node detection rate. However, greater collection of data regarding the usefulness of cervical sentinel node biopsy using indocyanine green is necessary.     

腺相关病毒-1介导增强型绿色荧光蛋白基因在大鼠角膜基质细胞体内外转染的研究

目的:探讨血清1型腺相关病毒(rAAV1)介导增强型绿色荧光蛋白(EGFP)基因体内、外转染大鼠角膜基质细胞后绿色荧光蛋白(GFP)的表达及转染率。方法:采用携带绿色荧光蛋白报告基因的血清1型腺相关病毒(rAAV1-EGFP)感染原代培养的SD大鼠角膜基质细胞,通过荧光显微镜和流式细胞仪观察和检测EGFP的表达及阳性率。建立大鼠异体角膜移植模型,用含rAAV1-EGFP转染液的培养基在37℃孵育SD大鼠角膜植片同时浸泡缝线18h,再用此缝线间断缝合,将植片移植到大鼠角膜植床,术后通过荧光体视镜观察Wistar大鼠角膜出现EGFP荧光的起始时间及分布情况。结果:按rAAV1-EGFP不同转染倍数(MOI)转染角膜基质细胞。转染后3d,在倒置荧光显微镜下观察到角膜基质细胞的细胞质有GFP阳性表达。体视荧光显微镜观察到大鼠角膜中开始有GFP阳性表达;随MOI值的增高而增强,转染后7~9d达到高峰,此时检测rAAV1-EGFP对角膜基质细胞的转染效率,MOI=103时是16.3%,MOI=104时是30.3%,MOI=105时是43.6%,MOI=106时是47.5%。rAAV1-EGFP在大鼠角膜中的表达也明显增强。结论:rAAV1-EGFP可以在体内外稳定、有效转染大鼠角膜基质细胞,是角膜基质细胞理想的报告基因。     

Neuroprotective effects of tongluojiunao in neurons exposed to oxygen and glucose deprivation

Ethnopharmacological relevance

TongLuoJiuNao (TLJN) is an herb extract that mainly contains ginsenoside Rg1 and geniposide, which are clinically used for treating ischemic damages in the brain.

Aim of the study

In the stroke, cerebral ischemia followed by oxygen reperfusion induced apoptosis in hippocampal neurons, while extension of axons and dendrites in neurons may compensate for and repair damages of neuronal network in the hypoxia brain. In this study, we investigated whether TLJN can protect neurons against damages by ischemia in brain vasculature.

Materials and methods

We measured cell viability and lactate dehydrogenase (LDH) release from primary culture of rat hippocampal neurons before and after the neurons were deprived of oxygen and glucose (OGD). In addition, the effects were evaluated with cell viability and neurite outgrowth before or after OGD.

Results

We found that TLJN could play a neuroprotective role to cultured primary rat hippocampal neurons under both normal and oxygen/glucose-deprivation (OGD) conditions. TLJN could protect both cultured primary rat hippocampal neurons and brain microvascular endothelial cells (BMECs) from cell death under both normal and oxygen/glucose-deprivation (OGD) conditions. Moreover, under the same conditions, BMECs-conditioned media pretreated by TNJN could also promote neuron viability and neurite outgrowth, indicating that TLJN stimulated BMECs to secret some neuroprotective/neurotrophic factors.

Conclusion

These findings suggest that TLJN has a marked neuroprotective and neurotrophic roles by either direct or indirect operation, and provide insight into the mechanism of clinical efficacy of this drug against stroke.     

Effects of Schisandra chinensis Baillon (Schizandraceae) on lipopolysaccharide induced lung inflammation in mice

Aim of the study

This study was aimed to evaluate the neuroprotective and anti-aging activity of extracts in Caenorhabditis elegans from the roots and leaves of Damnacanthus officinarum Huang to provide the pharmacological basis in traditional medicine.

Materials and methods

Investigations on the neuroprotective and lifespan activity were carried out, which were observed by utilizing the following models: observing the worms’ chemosensory behavior test based on the aversion index in the assay plate, neuroprotective activity of nematode by evaluating the ASH neuron survival and lifespan test in C. elegans.

Results

It has been shown that the ethanol, n-butanol and aqueous extracts in the roots possessed significantly neuroprotective effect both in chemosensory behavior test and ASH neuron survival model. The same extracts in the leaves showed similar activities in two models, but have less potency revealing by the data. Four candidate extracts, possessing excellent neuroprotective activity, extend lifespan in C. elegans. The n-butanol extracts in the root part showed best efficacy among them.

Conclusion

The results show the n-butanol and aqueous extracts are the major pharmacological plant extracts. Moreover, the neuroprotective and lifespan-extension activity effects of root extracts are superior to leave extracts, supporting the traditional application of above-ground parts of DOH in treating various diseases associated with brain disorders and anti-aging.     

The influence of liver dysfunction on cyclosporine pharmacokinetics —A comparison between 70 per cent hepatectomy and complete bile duct ligation in dogs—

The influence of experimentally induced hepatic dysfunction on the pharmacokinetics of Cyclosporine A (CsA) was determined in dogs. The pharmacokinetics of oral (PO) and intravenous (IV) CsA were studied before and after 70 per cent hepatectomy or complete bile duct ligation (CBDL). Changes in liver function were monitored by serial measurements of serum bilirubin, and by the maximum removal rate (Rmax) and plasma disappearance rate (ICG-K) of indocyanine green (ICG). Concentrations of CsA in whole blood were measured by HPLC. Seventy per cent hepatectomy caused significant liver dysfunction: the ICG-Rmax decreased by 47.7±7.1 per cent (mean±SD) and the ICG-K decreased by 61.3±9.7 per cent during the first week after hepatectomy. At the same time, the systemic clearance (CLs) of IV-CsA decreased by 43.9±8.2 per cent, the area under the concentration curve (AUC) of IV-CsA increased by 35.4±20.8 per cent and the bioavailability of CsA decreased by 26.4±14.8 per cent. CBDL also induced significant liver dysfunction: the ICG-Rmax decreased by 39.1±12.8 per cent and the ICG-K decreased by 65.6±3.6 per cent in the second week after the operation. During the same period, the AUC of PO-CsA decreased by 69.9±10.7 per cent and the bioavailability of CsA also decreased markedly by 73.9±15.6 per cent. These data indicate that hepatic impairment significantly influences the pharmacokinetics of CsA, not only by the changes in intestinal absorption, but also by those in hepatic, metabolism. Dose adjustment is therefore necessary in the presence of hepatic dysfunction in order to maintain an adequate blood concentration of CsA without causing side effects. This research was performed in the Department of Surgery, University of Pittsburgh Health Center, University of Pittsburgh, USA     

Comparison of the Clinical Manifestations between Acute Vogt-Koyanagi-Harada Disease and Acute Bilateral Central Serous Chorioretinopathy

PurposeTo compare clinical, angiographic, and optical coherence tomographic characteristics between eyes with acute Vogt-Koyanagi-Harada (VKH) disease and eyes with acute bilateral central serous chorioretinopathy (CSC), and to demonstrate distinguishing features between the two diseases in confusing cases.MethodsThe medical records of 35 patients with VKH disease and 25 patients with bilateral CSC were retrospectively reviewed. Characteristics according to slit-lamp biomicroscopy, ophthalmoscopy, fundus photography, fluorescein angiography, indocyanine green angiography, and spectral-domain optical coherence tomography were compared between the two diseases.ResultsFive of 35 patients (10 of 70 eyes, 14.3%) with VKH disease were initially misdiagnosed as CSC patients, and six of 25 patients (12 of 50 eyes, 24%) with bilateral CSC were initially misdiagnosed as patients with VKH disease. Pigment epithelial detachment in CSC and optic disc hyperemia in VKH disease show the highest positive predictive values of 100% for each disease.ConclusionsOptic disc hyperemia in VKH disease and pigment epithelial detachment in bilateral CSC are the most specific clinical manifestations of each disease at initial patient presentation.     

Recent advances in the treatment of hilar cholangiocarcinoma: portal vein embolization

The clinical application of portal vein embolization (PVE) has contributed to improving the postoperative outcome of hilar cholangiocarcinoma. The enlarged nonembolized lobe after PVE protects the patient from postoperative hepatic failure, due to the increased functional reserve, and shortens the hospital stay. Although numerous reports have shown beneficial effects of PVE on postoperative outcome after extended hepatectomy, no randomized controlled study has been performed so far. It is urgent to establish a “gold standard” of PVE, because the indications, approach to the portal vein, types of embolic materials, and methods used to evaluate the function of the future liver remnant are variable among institutions. The indications and procedures of PVE for hilar cholangiocarcinoma may be different from those for hepatocellular carcinoma or colorectal metastasis, because, in many patients with hilar cholangiocarcinoma, biliary cancer is associated with biliary obstruction and cholangitis. This review article summarizes the contribution of PVE to the outcome of postoperative management in patients with hilar cholangiocarcinoma needing extended hepatectomy. We also describe our PVE procedure, which has been established from our experience of more than 240 cases of biliary cancer. Furthermore, the drawbacks of PVE, which may reduce the pool of candidates for surgery, are also discussed.     

Sweet potato leaf extract inhibits the simulated in vitro gastrointestinal digestion of native starch

Several studies have reported the therapeutic use of caffeoylquinic acid (CQA) derivatives in the management of hyperglycemia. This study used a simulated in vitro gastrointestinal digestion model to assess the inhibitory effects of CQA derivatives-rich sweet potato leaf extract (SPLE) and a commercially produced green coffee bean extract (GCBE), each with total polyphenols contents of 452 mg g⁻¹ and 278 mg g⁻¹, respectively, against starch digestion. The changes in the amounts of total polyphenols and total CQA derivatives during in vitro gastrointestinal digestion were also examined. The results indicated that both extracts contained substantial levels of CQA derivatives (136 mg g⁻¹ and 83.5 mg g⁻¹ of extract for SPLE and GCBE, respectively). The amounts of total polyphenols and total CQA derivatives in 20 mg of SPLE and GCBE samples decreased from 9.04 mg to 0.58 mg and from 5.56 mg to 0.58 mg, and from 2.72 mg to 0.16 mg and from 1.67 mg to 0.10 mg, respectively, following in vitro gastrointestinal digestion and subsequent dialysis. When SPLE and GCBE were accompanied with starch for in vitro digestion test, they both exhibited inhibitory effect against starch digestion during simulated intestinal digestion, with estimated half maximal inhibitory concentration (IC₅₀) values of 4.91 mg and 6.06 mg polyphenols, respectively. The amount of glucose permeated through dialysis membrane also decreased significantly in comparison with the extract-negative control. Thus, both SPLE and GCBE were capable of modulating the release of glucose from starch digestion in simulated intestinal tract. The observed inhibitory effects against glucose release were presumably due in part to the presence of CQA derivatives in the tested extracts. The SPLE had higher inhibitory effect against in vitro starch digestion than the commercially prepared reference GCBE. Therefore, the SPLE might be used to manage hyperglycemia over the long term.     

Regulation of cell growth and expression of 7B2, PC2, and PC1/3 by TGFbeta 1 and sodium butyrate in a human pituitary cell line (HP75)

Recent studies have shown that 7B2 and the neuroendocrine-specific proconvertase PC2 have important roles in pituitary cell proliferation and hormone secretion. Studies from our laboratory have also shown that TGFb1 regulates anterior pituitary cell proliferation and hormone secretion. To study the regulation of 7B2 in human pituitary tumors, we used a cell line derived from a human pituitary adenoma (HP75) that has been shown to express 7B2, PC1, PC2, and TGFβ receptors to analyze the effects of TGFβ1 and the histone deacetylase inhibitor (HDACI) sodium butyrate (NaB) treatment on 7B2 mRNA expression along with the neuroendocrine-specific proconvertases 1/3 (PC1) and PC2 mRNA and protein expression. RNA was quantified by real-time PCR and proteins were detected by immunohistochemistry and Western blotting. Treatment of cells with 1 mM NaB or 1 nM TGFβ1 for 4 d decreased cell proliferation with a concomitant increase in the cell cycle protein p21. Real-time PCR analysis showed a significant increase in 7B2 mRNA after NaB and TGFβ1 treatment. PC2 mRNA was down regulated by NaB while PC1 mRNA was unchanged. TGFβ1 stimulated PC1, but not PC2 mRNA levels. Changes in PC1 and PC2 protein were similar to changes in the mRNAs, but the differences were not significant. These results indicated that NaB and TGFβ1 inhibit pituitary cell proliferation and regulate the expression of 7B2, PC1, and PC2 in a cell culture model of pituitary tumors. Our results also indicate that inhibition of pituitary cell proliferation is associated with increased expression of 7B2 mRNA.     

Clinical staining of the ocular surface: Mechanisms and interpretations

In this article we review the mechanism of ocular surface staining. Water-soluble dyes are excluded from the normal epithelium by tight junctions, the plasma membranes and the surface glycocalyx. Shed cells can take up dye. A proportion of normal corneas show sparse, scattered time-dependent, punctate fluorescein uptake, which, we hypothesise, is due to a graded loss of the glycocalyx barrier, permitting transcellular entry into pre-shed cells. In pathological staining, there is little evidence of ‘micropooling’ at sites of shedding and the term ‘punctate erosion’ may be a misnomer. It is more likely that the initial event involves transcellular dye entry and, in addition, diffusion across defective tight junctions. Different dye-staining characteristics probably reflect differences in molecular size and other physical properties of each dye, coupled with differences in visibility under the conditions of illumination used. This is most relevant to the rapid epithelial spread of fluorescein from sites of punctate staining, compared to the apparent confinement of dyes to staining cells with dyes such as lissamine green and rose bengal. We assume that fluorescein, with its lower molecular weight, spreads initially by a paracellular route and then by transcellular diffusion. Solution-Induced Corneal Staining (SICS), related to the use of certain contact lens care solutions, may have a different basis, involving the non-pathological uptake of cationic preservatives, such as biguanides, into epithelial membranes and secondary binding of the fluorescein anion. It is transient and may not imply corneal toxicity. Understanding the mechanism of staining is relevant to the standardisation of grading, to monitoring disease and to the conduct of clinical trials.