Dietary Green Tea Extract and Antioxidants Improve Insulin Secretory Functions of Pancreatic β-Cells in Mild and Severe Experimental Rodent Model of Chronic Pancreatitis

Chronic pancreatitis (CP) is a progressive inflammatory disorder of the pancreas. It is predominantly idiopathic (with an unknown cause) in India and mostly due to alcohol in the West. Diabetes that occur secondary to chronic pancreatitis (T3c Diabetes) is often brittle, and is difficult to attain normoglycemia with conventional treatment requiring multiple doses of insulin. Mild and severe model of CP was induced in mice by repeated intraperitoneal injections of cerulein and L-arginine respectively with an intent to study islet dysfunction and develop therapeutic strategy in animal models of CP. Dietary intervention of epigallocatechin-3-gallate (EGCG) was tested in both the models of CP for its beneficial effects on insulin secretory functions. Pancreata collected upon euthanasia were used to study alterations in the morphology of pancreatic parenchyma and inflammation by staining with H&E and fibrotic changes by Masson’s trichrome and picrosirius staining. Insulin secretory functions of islets were evaluated to test the efficacy of the dietary intervention on β-cell functions. Intraperitoneal glucose tolerance test was performed to monitor the glucose homeostasis before and after the dietary intervention. Both the models resulted in CP with dispersed acini, inflammation and fibrosis. The loss of acini and extent of fibrosis was more in L-arginine model. 2-fold improvement in glucose-stimulated insulin secretory functions of islets was observed with 0.5% EGCG dietary intervention in cerulein model of CP and 1.6-fold in L-arginine model of CP. A further improvement in insulin secretion by 3.2-fold was observed with additional dietary supplements like N-acetyl cysteine, curcumin in combination with EGCG. Our results thus demonstrate and highlight the therapeutic potential of dietary green tea (EGCG) supplementation in reversing islet dysfunction and improving glucose homeostasis in experimental chronic pancreatitis in mice.

Chronic spinal cord compression associated with intervertebral disc degeneration in SPARC-null mice

Chronic spinal cord compression(CSCC)is induced by disc herniation and other reasons,leading to movement and sensation dysfunction,with a serious impact on quality of life.Spontaneous disc herniation rarely occurs in rodents,and therefore establishing a chronic spinal cord compression(CSCC)animal model is of crucial importance to explore the pathogenesis and treatment of CSCC.The absence of secreted protein,acidic,and rich in cysteine(SPARC)leads to spontaneous intervertebral disc degeneration in mice,which resembles human disc degeneration.In this study,we evaluated whether SPARC-null mice may serve as an animal model for CSCC.We performed rod rotation test,pain threshold test,gait analysis,and Basso Mouse Scale score.Our results showed that the motor function of SPARC-null mice was weakened,and magnetic resonance images revealed compression at different spinal cord levels,particularly in the lumbar segments.Immunofluorescence staining and western blot assay showed that the absence of SPARC induced apoptosis of neurons and oligodendrocytes,activation of microglia/macrophages with M1/M2 phenotype and astrocytes with A1/A2 phenotype;it also activated the expression of the NOD-like receptor protein 3 inflammasome and inhibited brain-derived neurotrophic factor/tyrosine kinase B signaling pathway.Notably,these findings are characteristics of CSCC.Therefore,we propose that SPARC-null mice may be an animal model for studying CSCC caused by disc herniation.

Classification and Nomenclature of Plant Metallothionein-like Proteins Based on Their Cysteine Arrangement Patterns

随着植物基因组研究的进展 ,在基因文库和蛋白文库登录的植物类金属硫蛋白基因已超过 5 0个 ,接近金属硫蛋白总数的 1/ 3,而且有不断上升的趋势。鉴于目前植物类金属硫蛋白命名与分类随意性太大 ,很有必要建立一个统一合理的命名与分类法。对植物类金属硫蛋白一级结构进行详细分析后 ,发现该蛋白两端富含半胱氨酸的区域内半胱氨酸的排列方式颇具规律性 ,进而提出了以半胱氨酸排列方式为基础的分类及命名法 ,并阐述了采用这种方法的理由及其可行性。

Application of L-Cys-Capped CdS Nanoparticles in Determination of Nucleic Acid by Resonance Light-Scattering Technique

Aim To determine nucleic acid （DNA） using Nanometer-sized L-cysteine-capped CdS particles by resonance light scattering （RLS） method. Methods The nano-particles synthesized by a colloidal aqueous method were water-soluble, stable, and highly luminescent. The RLS of L-Cys-CdS particles were greatly quenched by DNA in Tris-HCl solutions. The intensity of RLS at 344 nm was proportional to the concentration of DNA. Results The linearity range of the calibration curve was 0. 01 - 1.0 μg·mL^-1 for calf thymus DNA and 0. 04 - 1.5 μg· mL^-1 for salmon sperm DNA. The detection limits （3 δ） were 8 ng·mL^-1 for calf thymus DNA and 10 ng·mL^-1 for salmon sperm DNA. Conclusion This method is simple, sensitive, and capable of avoiding the use of toxic dyes.

小鼠CSRP2基因的克隆、融合表达及蛋白的分离纯化

目的:获得小鼠CSRP2(cysteine rich protein 2)基因片段,高效表达、纯化CSRP2。方法:用RT-PCR技术从小鼠心脏组织中提取总DNA并扩增出相应大小的CSRP2 DNA片段,将其与pGEM-T-easy载体连接后测序,将测序正确的CSRP2从BamHⅠ和HindⅢ酶切位点克隆入原核表达载体pRSET-A,将连接产物转化大肠杆菌BL21,挑出阳性克隆,IPTG诱导表达重组的6×His融合蛋白,对重组融合蛋白通过镍柱进行纯化。结果与结论:PCR获得的CSRP2序列与GenBank报道的一致(为675 bp);重组载体在大肠杆菌BL21中以包涵体形式高效表达;融合蛋白经SDS-PAGE和Western blot分析,在相对分子质量约24×103处有特异的蛋白条带,目的蛋白表达量占菌体总蛋白量的25%;重组蛋白经镍柱纯化后,得到了高纯度的CSRP2融合蛋白。

半胱氨酸蛋白酶激动剂、X连锁凋亡抑制蛋白、半胱氨酸天冬氨酸蛋白酶3在鼻腔鼻窦鳞状细胞癌中的表达

现代肿瘤学认为肿瘤的发病与细胞凋亡数量的减少密切相关,第二个线粒体衍生的半胱氨酸蛋白酶激动剂（second mitochondria-derived activator of caspase,Smac）、X连锁凋亡抑制蛋白（X-linked inhibitor of apoptosis protein,XIAP）、半胱氨酸天冬氨酸蛋白酶3（cysteine containing aspartate specific protease,caspase-3）在细胞凋亡过程中起着重要的调控及效应作用。本研究采用免疫组化方法观察三者在鼻腔鼻窦鳞状细胞癌（nasal squamous cellcarcinoma,NSCC）及不同病理分期中的表达情况。

理想肌松药的历史与现状

自从箭毒引入临床麻醉，外科手术学得到迅猛发展，但其残留肌松作用等副作用增加了患者风险，因此人们一直在寻求和研制理想的肌松药。近年正在研制的gantacurium和AV002有可能成为理想肌松药。而sugammadex和cysteine则符合理想拮抗剂的条件。

Inhibition of cysteine protease papain by metal ions and polysulfide complexes,especially mercuric ion

Aim Cysteine proteases are closely associated with many human and non-human pathological processes and are potential targets for metal ions especially Hg^2＋ and the related species. In the present work, on the basis of to the general study on the effects of some metal ions on the activity of papain, a well-known representative of cysteine protease family, the inhibitory effects of Hg^2＋ and polysulfide complexes were studied. Results All the metal ions tested （Hg^2＋, Cu^2＋, Ag^＋, Au^3＋, Zn^2＋, Cd^2＋, Fe^3＋, Mn^2＋, Pb^2＋, Yb^3＋） inhibit the activity of papain anda good correlation between the inhibitory potency and softness-and-hardness was observed. Among the metals, Hg^2＋ was shown to be a potent inhibitor of papain with a Kiof 2 × 10^-7 mol·L^-1 among. Excessive amounts of glutathione and cysteine could reactivate the enzyme activity of papain deactivated by Hg^2＋. These evidences supported that Hg^2＋ might bind to the catalytic site of papain. Interestingly, Hg （Ⅱ） polysulfide complexes were for the first time found to inhibit papain with a Ki of 7 × 10^-6 mol·L^-1, whose potency is close to a well known mercury compound, thimerosal （Ki=2.7 × 10^-6）. In addition, Hg （Ⅱ） polysulfide complexes exhibit good permeability （ 1.9 × 10-5 cm· s^-1） to caco-2 monolayer. Conclusion These results suggested that mercury polysulfide complexes might be potential bioactive species in the interaction with cysteine proteases and other- SH-content proteins, providing a new clue to understand the mechanism of the toxicological and pharmacological actions of cinnabar and other insoluble mercury compounds.

Modeling the Cysteine Rich Domain of Plant Metallothionein-like Protein

With the progress of plant genome research, more than 50 plant metallothionein_like (MT_L) genes have been found, but only several MT_L proteins have been detected and no experimental structural information for MT_L proteins has been reported so far. Since detailed knowledge of the protein tertiary structure is required to understand its biological function, a method is needed to determine the structure of these proteins. In this study, the structural data of known mammal MT was used to determine the interatomic distance constraints of the CXC and CXXC motifs and the metal_sulfur chelating cluster. Then several possible MT conformations were predicted using a distance geometry algorithm. The statistical analysis was used to select those with much lower target function values and lower conformation energies as the predicted tertiary structural models of the cysteine_rich (CR) domains of these proteins. A suitable prediction method for modeling the CR domain of the plant MT_L protein was constructed. The accurately predicted result for the known structure of an MT protein from blue crab suggests that this method is practicable. The tertiary structures of CR domains of rape MT_L protein LSC54 was then modeled with this method.

Novel Strategy for Synthesis of High Quality CdTe Nanocrystals in Aqueous Solution

A novel aqueous route for the synthesis of high-quality CdTe nanocrystals（NCs） is presented in this article. With both glutathione（GSH） and cysteine[n（GSH）：n（cysteine）=1：3] as stabilizers, high-quality CdTe NCs with controllable photoluminescence（PL） wavelength from 500 to 630 nm can be prepared within 4 h. As-prepared CdTe NCs show higher photoluminescence quantum yields（PLQY） compared with CdTe NCs prepared via other aqueous methods. When the fluorescent emission peak appeared in orange-red window, the PLQY reaches 70% or above at room temperature without any post-preparative treatment.

Correlation of matrix metalloproteinase suppressor genes RECK,VEGF,and CD105 with angiogenesis and biological behavior in esophageal squamous cell carcinoma

AIM: To explore the expression of reversion inducing cysteine-rich protein with Kazal motifs (RECK), vascular endothelial growth factor (VEGF) and endoglin (CD105) protein and its correlation with occurrence, development, invasion and metastasis in esophageal squamous cell carcinoma (ESCC). METHODS: Streptavidin-peroxidase (SP) immunohisto- chemistry was used to detect expression of RECK and VEGF in 62 cases of ESCC, 31 cases of adjacent atypical hyperplastic epithelium and 62 cases of normal esophageal epithelium. CD105 Mb was used to assess microvessel density (MVD). RESULTS: The expression of RECK was closely correlated with histological grade, infiltrative depth and lymphatic metastasis in ESCC (P < 0.05). The expression of RECK decreased during cancer development: normal esophageal epithelium (85.5%, 53/62), adjacent atypical hyperplastic epithelium (71.0%, 22/31), and carcinoma (59.7%, 37/62). There was a significant difference among the groups (P < 0.05). The expression of VEGF protein was closely correlated with infiltrative depth and lymphatic metastasis in ESCC (P < 0.05). The expression of VEGF protein increased during cancer development: normal esophageal epithelium (29.0%, 18/62), adjacent atypical hyperplastic epithelium (54.8%, 17/31), and carcinoma (67.7%, 42/62). There was a significant difference among the groups (P < 0.05). MVDCD105 increased in accordance with histological grade, butthere was no significant difference (grade Ⅰ, 36.92 ± 10.85; grade Ⅱ, 37.65 ± 9.50; and grade Ⅲ, 38.06 ± 12.19). The MVDCD105 was closely correlated with infiltration and lymphatic metastasis in ESCC (P < 0.05). The expression of RECK was inversely correlated with the expression of VEGF and CD105. CONCLUSION: RECK, VEGF and CD105 play important roles in the infiltration, metastasis and carcinogenesis in esophageal carcinoma. Angiogenesis in ESCC may be promoted by over-expression of CD105.

Synergistic effects of elevated homocysteine level and abnormal blood lipids on the onset of stroke

Hyperhomocysteinemia and abnormal blood lipids are independent risk factors for stroke. However, whether both factors exert a synergistic effect in the onset of stroke remains unclear. The present study is a retrospective analysJs of 2 089 cases of stroke and 2 089 control cases of simple in- tervertebral disk protrusion using a paired multivariate logistic regression method. Adjusting for known confounding variables including the patients＇ age, gender, smoking status, alcohol con- sumption status, patient and family medical history, and clinical biochemical indices, elevated ho- mocysteine level was related to the onset of stroke. Patients with elevated homocysteJne levels and abnormal blood lipids showed a 40.9 % increase in the risk for stroke compared to patients with normal homocysteine levels and blood lipids （odds ratio 1.409; 95% confidence interval 1.127-1.761）. These results indicate that elevated homocysteine and abnormal blood lipids exert synergistic effects in the onset of stroke. Patients with elevated homocysteine levels and abnormal blood lipids are predisposed to stroke.

Quality of healing of gastric ulcers: Natural products beyond acid suppression

Gastric ulcer is a chronic disease featured with unexpected complications, including bleeding, stenosis and perforation, as well as a high incidence of recurrence. Clinical treatments for gastric ulcer have allowed the rapid development of potent anti-ulcer drugs during the last several decades. Gastric ulcer healing is successful with conventional treatments including H2-receptor antagonists, and proton pump inhibitors(PPIs) have been essential for ulcer healing and prevention of complications. Additionally, Helicobacter pylori eradication therapy is effective in reducing ulcer recurrence and leads to physiological changes in the gastric mucosa which affect the ulcer healing process. However, in spite of these advancements, some patients have suf-fered from recurrence or intractability in spite of continuous anti-ulcer therapy. A new concept of the quality of ulcer healing(QOUH) was initiated that considers the reconstruction of the mucosal structure and its function for preventing ulcer recurrence. Although several gastroprotection provided these achievements of the QOUH, which PPI or other acid suppressants did not accomplish, we found that gastroprotection that originated from natural products, such as a newer formulation from either Artemisia or S-allyl cysteine from garlic, were very effective in the QOUH, as well as improving clinical symptoms with fewer side effects. In this review, we will introduce the importance of the QOUH in ulcer healing and the achievements from natural products.

头孢米诺钠(美士灵)的基础与临床研究进展

头孢米诺（cefminox）其钠盐供注射用,商品名为美士灵（meicelin）,系明治制果中央研究所从放线菌发酵液中发现的,其药效学、药动学及临床应用进展简介如下: 头孢米诺钠的分子式为C16H20N7NaO7S3·7H2O,分子量为667.67。它是7β位侧链末端有D—氨基酸（D-cysteine）的头孢菌素类抗生素。 抗菌谱:本品对革兰阳性、阴性菌均显示抗菌作用,尤其对链球菌（肠球菌除外）,大肠杆菌,肺炎杆菌,变形杆菌。

Cytoprotective effects of amifostine,ascorbic acid and N-acetylcysteine against methotrexate-induced hepatotoxicity in rats

AIM: To investigate the potential role of oxidative stress and the possible therapeutic effects of N-acetyl cysteine (NAC), amifostine (AMF) and ascorbic acid (ASC) in methotrexate (MTX)-induced hepatotoxicity.

Selectively colorimetric detection of cysteine with triangular silver nanoprisms

Triangular silver nanoprisms were prepared and applied to make colorimetric detection of cysteine based on our findings that cysteine could lead to the blue shift of the dipole plasmon resonance absorption, but other 19 kinds of natural amino acids could not. Cysteine with a concentration 160 nmol/L can result in a color change that can be discerned with naked eyes.

Aberrant methylation of secreted protein acidic and rich in cysteine gene and its significance in gastric cancer

BACKGROUND Aberrant methylation in DNA regulatory regions could downregulate tumor suppressor genes without changing the sequences.However,our knowledge of secreted protein acidic and rich in cysteine(SPARC)and its aberrant methylation in gastric cancer(GC)is still inadequate.In the present research,we performed fundamental research to clarify the precise function of methylation on SPARC and its significance in GC.AIM To investigate promoter methylation and the effects of the SPARC gene in GC cells and tissues and to evaluate its clinical significance.METHODS Plasmids that overexpressed the SPARC gene were transfected into human GC BGC-823 cells;non-transfected cells were used as a control group(NC group).Quantitative real-time polymerase chain reaction and western blotting(WB)were then used to detect the expression of SPARC.Methylation-specific polymerase chain reaction was executed to analyze the gene promoter methylation status.Cell viability was measured by the cell counting kit-8 assay.The migration and invasion ability of cells were detected by scratch assays and transwell chamber assays,respectively.Cell cycle events and apoptosis were observed with a flow cytometer.RESULTS The expression of SPARC mRNA in GC tissues and cells was significantly lower and showed differing degrees of hypermethylation,respectively,than that in normal adjacent tissues and control cells.Treatment with 5-Aza-2’-deoxycytidine(5-Aza-Cdr)was able to restore the expression of SPARC and reverse promoter hypermethylation.Overexpression of the SPARC gene significantly inhibited proliferation,migration,and invasion of GC cells,while also causing cell cycle arrest and apoptosis;the NC group exhibited the opposite effects.CONCLUSION This study demonstrated that SPARC could function as a tumor suppressor and might be silenced by promoter hypermethylation.Furthermore,in GC cells,SPARC inhibited migration,invasion,and proliferation,caused cell cycle arrest at the G0/G1 phase,and promoted apoptosis.

Shattering the castle walls: Anti-stromal therapy for pancreatic cancer

Despite the availability of potent chemotherapy regimens, such as 5-fluorouracil, folinic acid, irinotecan, and oxaliplatin(FOLFIRINOX) and nab-paclitaxel plus gemcitabine, treatment outcomes in metastatic pancreatic cancer(PC) remain unsatisfactory. The presence of an abundant fibrous stroma in PC is considered a crucial factor for its unfavorable condition. Apparently, stroma acts as a physical barrier to restrict intratumoral cytotoxic drug penetration and creates a hypoxic environment that reduces the efficacy of radiotherapy. In addition, stroma plays a vital supportive role in the development and progression of PC, which has prompted researchers to assess the potential benefits of agents targeting several cellular(e.g., stellate cells) and acellular(e.g., hyaluronan) elements of the stroma. This study aims to briefly review the primary structural properties of PC stroma and its interaction with cancer cells and summarize the current status of antistromal therapies in the management of metastatic PC.

The Electrochemical Behavior of Au/Au NPs/PNA/ZnS e-QD/ACA Electrode Towards Cy SH Oxidation

This work describes the electrochemical behavior of azodicarboxamide(ACA) film immobilized on the surface of penicillamine(PNA)/Zn Se-quantum dot(Zn Se-QD) gold nanoparticle(Au NPs) Au electrode. Electrocatalytic activity of modified electrode toward the oxidation of cysteine(Cy SH) was investigated. The surface structure and composition of the sensor were characterized by scanning electron microscopy(SEM). Oxidation of Cy SH on the surface of modified electrode was investigated with cyclic voltammetry, electrochemical impedance spectroscopy(EIS),hydrodynamic voltammetry and chronoamperometry methods. The results show that the PNA/Zn Se-QD/ACA film displays excellent electrochemical catalytic activities towards Cy SH oxidation. The modified electrode shows reproducible behavior and high level of stability during the electrochemical experiments. Also it has short response time, low detection limit, high sensitivity and low operation potential, which can be used as an amperometric sensor for monitoring of Cy SH. The proposed modified electrode was successfully used for determination of Cy SH in real sample such as human serum.

JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by Z-VAD.fmk and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells.