甘露糖敏感性绿脓杆菌制剂对加热后肝癌细胞生物学特性的影响

目的 探讨甘露糖敏感性绿脓杆菌制剂（PA-MSHA）对水浴加热后有转移潜能的人肝癌MHCC97L细胞增殖和侵袭能力的影响及其机制。方法 42℃水浴加热前2 h加入1×10^11/L PA-MSHA,加热组不加药,对照组不加热、不加药,其他处理相同并在同一时间点观察。在不同的时间观察细胞增殖、生长曲线、克隆形成率、流式细胞术分析细胞周期（FCM）、侵袭、运动、酶联免疫吸附试验（ELISA）法检测血管内皮生长因子（VEGF）和基质金属蛋白酶（MMP）-2蛋白表达。结果PA-MSHA能够逆转加热对人肝癌MHCC97L细胞的增殖促进作用,和对照组比较,加热联合PA-MSHA能明显抑制人肝癌MHCC97L细胞的增殖（P〈0.01）,其最大抑制效应为加热后48 h（42.3%）,细胞倍增时间延长了1.68倍,加热联合PA-MSHA组其加热后48、96 h的细胞集落形成率均明显降低（P〈0.01）,FCM显示,加热联合PA-MSHA能逆转加热对细胞周期的影响,和对照组比较,其48 h的G1期细胞比例明显升高,S＋G2期细胞比例明显降低（P〈0.01）,其96 h的G1期和S＋G2期细胞比例差异无统计学意义（P〈0.05）。加热联合PA-MSHA组其加热后48 h和96 h相同数量的细胞穿过人工基底膜到达Transwell小室膜背面的细胞平均数（侵袭实验）和穿过Transwell小室膜到达背面的MHCC97L细胞平均数（运动实验）均明显低于加热组（P〈0.01）,ELISA法检测相同数量细胞的上清液发现,加热联合PA-MSHA组其加热后48、96 h细胞VEGF和MMP-2蛋白的分泌量和加热组比较差异无统计学意义（P〈0.05）。结论 PA-MSHA能抑制体外加热后肝癌细胞的增殖,其作用和抑制细胞进入DNA合成期和分裂期有关,进一步抑制其侵袭运动能力,其作用和MMP-2、VEGF表达无关。

基于对巯基苯硼酸的糖敏感水凝胶的制备

本文通过简单直接混合海藻酸钠、对巯基苯硼酸和Ca Cl2构建水凝胶。通过扫描电子显微镜观察到水凝胶具有明显的大孔多孔结构。溶胀实验证实随着对巯基苯硼酸浓度的增加,水凝胶的稳定性逐渐增强。此外,随着溶液中葡萄糖浓度的增加,凝胶降解速率逐渐加快。该多孔糖敏感水凝胶有望作为药物载体用于糖尿病的治疗。

铜绿假单胞菌制剂在恶性肿瘤中的辅助治疗作用

当前,恶性肿瘤已越来越趋向于多方面的综合治疗.带有甘露糖敏感性血凝菌毛的铜绿假单胞菌(pseudomonas aeruginosa-mannosesensitive hemagglutinin,PA-MSHA)制剂是一类用于免疫治疗的生物制剂,正被应用于恶性肿瘤的治疗过程中.本文重点介绍PA-MSHA制剂对恶性肿瘤辅助治疗作用的研究进展.

Glucose-responsive artificial promoter-mediated insulin gene transfer improves glucose control in diabetic mice

AIM:To investigate the effect of insulin gene therapy using a glucose-responsive synthetic promoter in type 2 diabetic obese mice.METHODS:We employed a recently developed novel insulin gene therapy strategy using a synthetic promoter that regulates insulin gene expression in the liver in response to blood glucose level changes.We intravenously administered a recombinant adenovirus expressing furin-cleavable rat insulin under the control of the synthetic promoter(rAd-SP-rINSfur) into diabetic Lepr db/db mice.A recombinant adenovirus expressing β-galactosidase under the cytomegalovirus promoter was used as a control(rAd-CMV-βgal).Blood glucose levels and body weights were monitored for 50 d.Glucose and insulin tolerance tests were performed.Immunohistochemical staining was performed to investigate islet morphology and insulin content.RESULTS:Administration of rAd-SP-rINSfur lowered blood glucose levels and normoglycemia was maintained for 50 d,whereas the rAd-CMV-βgal control virus-injected mice remained hyperglycemic.Glucose tolerance tests showed that rAd-SP-rINSfur-treated mice cleared exogenous glucose from the blood more efficiently than control virus-injected mice at 4 wk [area under the curve(AUC):21 508.80 ± 2248.18 vs 62 640.00 ± 5014.28,P < 0.01] and at 6 wk(AUC:29 956.60 ± 1757.33 vs 60 016.60 ± 3794.47,P < 0.01).In addition,insulin sensitivity was also significantly improved in mice treated with rAd-SP-rINSfur compared with rAd-CMV-βgal-treated mice(AUC:9150.17 ± 1007.78 vs 11 994.20 ± 474.40,P < 0.05).The islets from rAd-SP-rINSfur-injected mice appeared to be smaller and to contain a higher concentration of insulin than those from rAd-CMV-βgal-injected mice.CONCLUSION:Based on these results,we suggest that insulin gene therapy might be one therapeutic option for remission of type 2 diabetes.

Effects of glucocorticoids on the growth and chemosensitivity of carcinoma cells are heterogeneous and require high concentration of functional glucocorticoid receptors

AIM： To determine how glucocorticoids （GCs） may affect the growth and chemosensitivity of common carcinoma cells. METHODS： The effect of dexamethasone （DEX） on growth and chemosensitivity was assessed in 14 carcinoma cell lines. The function of GC receptors （GR） was assessed by MMTV reporter assay. Overexpression of GR was done by in vitro transfection and expression of a GR-expressing vector. Immunohistochemical stain of tissues and ceils were done by PA1-511A, an anti-GR monodonal antibody. RESULTS： DEX inhibited cell growth of four （MCF-7, MCF- 7/MXR1, MCF-7/TPT300, and HeLa）, increased cisplatin cytoxicity of one （SiHa）, and decreased dsplatin cytotoxicity of two （H460 and Hep3B） cell lines. The GR content of the seven cell lines affected by DEX was significantly higher than those of the seven cell lines unaffected by DEX （5.2±2.5×10^4 sites/cell vs1.3±1.4×10^4 sites/cell, P= 0.005）. Only two DEX-unresponsive cell lines {NPC-TW01 and NPC- TW04） oontained high GR amounts in the range （1.9-8.1×10^4 sites/cell） of the seven DEX-responsive cell lines. The GR function of NPC-TW01 and NPC-TW04, however, was foundto be impaired. The importance of high cellular amount of GR in mediating DEX susceptibility of the cells was further exemplified by GR dose-dependent drug resistance to cisplatin of AGS, a cell line with low GR content and was unaffected by DEX before transfection of GR-expressing vector. Immunohistochemical studies of human cancer tissues showed that 5 of the 45 （11.1%） breast cancer and 43 of the 85 （50.6%） non-small cell lung cancer had high GR contents at the ranges of the GC-responsive carcinoma cell lines. CONCLUSION： The growth and chemosensitivity of human carcinomas with high GR contents may be affected by GC. However, in light of the heterogeneous and even contradictive effects of GC on these cells, routine examination of GR contents of human carcinoma tissues may not be clinically useful until other markers that help predict the ultimate effect of GC on individual patients are identified.

EFFECT OF CHRONIC ACE INHIBITIONONGLUCOSE TOLERANCE AND INSULIN SENSITIVITY IN HYPERTENSIVE TYPE 2 DIABETIC PATIENTS

We studied 14 moderately overweight Typo 2 diabetic patients with essential hypertension in stable metabolic control after a run-in period , and again after 3 months of antihypertensive treatment with the angiotensin-converting enzyme (ACF) inhibitor captopril. Glucose tolerance was tested with a 75g oral glucose load (OGTT) and insulin sensitivity was measured by the insulin suppression test (IST) while dietary and drug treatment of the hyperglycemia was maintained constant. In the whole group. mean blood pressure (MBP) fell progressively over 3months from a baseline value of 123± 3 mmHg (1 mmHg= 0. 133 kpa) to a final value of 115± 2 mmHg(P<0. 005). After treatment, fasting plasma glucose, insulin, free fatty acid (FFA). potassium, and glycosylated hemoglobin concentrations were unchanged from baseline. There were no significant differences in glucose tolerance and insulin sensitivity between pre- and post-trearment values. Neither endogenous (oral glucose) nor exogenous (IST) insulin caused any change in plasma potassium concentration. This resistance to the hypokalemic action of insulin was not affected by captopril.

Measurement of β-1,3 Glucanase Activity in Permeabilized Discs of Leaves of Healthy and Scald-Diseased Plants

Leaf discs of five cultivars of sugarcane exhibiting different degree of susceptibility to leaf scald were used to measure β-1,3 glucanase activity before and after experimental infection with Xanthomonas albilineans. Leaf discs were permeabilized with iso-propanol to facilitate the uptake of the enzyme substrate by intact tissues and to improve the enzyme assay. Bacterial infection significantly enhances β-1,3 glucanase activity of sensitive cultivars whereas significantly decreased that of the resistant one. Low concentrations of salicylate increase the hydrolase activity whereas jasmonic acid do not act as an elicitor of the enzyme and β-1,3 glucanase, such as laminarin, significantly inhibits the production of β-1,3 glucanase. Thus, the enzyme must be considered as a sensitivity factor induced by the pathogen.

Antihyperglycemic, insulinsensitizing and antioxidant activities of the active fractions from Anoectochilus chapaensis

Anoectochilus chapaensis Gagnep. （Orchidaceae）, an indigenous and valuable Chinese folk medicine, has been widely used in China to treat diabetes. However, few reports are available about its constituents and activity. The present experiment was conducted to investigate the active fractions from A. chapaensis in diabetic rat model induced by high-fat diet plus streptozotocin. The total EtOH extract from the whole herbs ofA. chapaensis, half of which was partitioned in sequence with petrol ether （PE）, ethyl acetate （EtOAc）, n-BuOH and H2O, thus yielding four fractions, all of them were orally administered with an identical dose amount to 4 g/kg dried crude herbs once a day for consecutive two weeks to further investigate the antihyperglycemic activity. The EtOAc fraction caused a significant fall in the non-fasting blood glucose level of diabetic rats from （402.66±82.26） to （226.26±62.10） mg/dl, which may be attributed to ameliorating insulin resistance, modulating the activity of enzymatic antioxidants, reducing the content of NO, etc. Much more intact β cells in the islets of Langerhans in EtOAc fraction-treated groups than the negative control were observed, which greatly supported the morphological and functional elucidation. The OGTT evidenced that EtOAc fraction could promote the endurance capacity of acute glucose increase in diabetic rats. The EtOAc fraction of A. chapaensis contains some hypoglycemic and antioxidant principles with the potential to be developed further for the treatment of diabetes specifically associated with an insulin resistance state.

Treatment of cervical cancer by si RNA-loaded chitosan-coated calcium phosphate nanoparticles

In the present study, a chitosan-coated calcium phosphate nanoparticle（CS/CaP/siRNA NP） was developed to deliver si RNA for treatment of cervical cancer. The CS/CaP/siRNA NPs were prepared by the nano-precipitation method. The resulted NPs had a uniform spherical morphology with a size of ～194 nm and a zeta potential of ～＋27 mV. In vitro experiments demonstrated that these NPs could efficiently deliver si EGFR into Hela cells and significantly down-regulate the EGFR expression level, which was probably associated with enhanced cell adhesion of chitosan, leading to extended residence time of cell internalization. Then the internalized CS/CaP/siRNA NPs exhibited pH-responsive disassembly of NPs, resulting in the enhanced release of si RNA and rapid lysosomal escape into cytoplasm. Moreover, in vivo anticancer results showed that the CS/CaP/siRNA NPs had significant inhibitory effects on tumor growth after intratumoral injection in Hela tumor xenografted nude mice, accompanying with no obvious changes of body weight during the whole experimental period. All these results indicated that the CS/CaP/siRNA NPs would have great potential to deliver si RNA for the treatment of cervical cancer via mucosal administration.

New approach for chiral separation: from polysaccharide-based materials to chirality-responsive polymers

Chiral separation that is closely related to daily life is a meaningful research. Polysaccharide-(e.g., cellulose, amylose derivatives) based chiral packing materials afford powerful chiral stationary phases(CSPs) toward a broad range of racemic compounds. However, considering the explosive growth of specific chiral drugs, the separation efficiencies of these CSPs need further improvement, which calls for new approaches and strategies. Smart polymers can change their physical or chemical properties dynamically and reversibly according to the external stimuli(e.g., thermo-, pH, solvent, ion, light, critical parameters for chromatographic separation) exerted on them, subsequently resulting in tunable changes in the macroscopic properties of materials. In addition to their excellent controllability, the introduction of chiral characteristics into the backbones or side-chains of smart polymers provides a promising route to realize reversibly conformational transition in response to the chiral analytes. This dramatic transition may significantly improve the performance of materials in chiral separation through modulating the enantioselective interactions between materials and analytes. With the help of chirality-responsive polymers, intelligent and switchable CSPs could be developed and applied in column-liquid chromatography. In these systems, the elution order or enantioselectivity of chiral drugs can be precisely modulated, which will help to solve many challenging problems that involve complicated enantiomers. In this paper we introduce some typical examples of smart polymers that serve as the basis for a discussion of emerging developments of CPSs, and then briefly outline the recent CSPs based on natural and certain synthetic polymers.

A correlation study of the expression of resistin and glycometabolism in muscle tissue after traumatic brain injury in rats

Objective： To investigate the expression pattern of resistin （RSTN） in skeletal muscle tissue and its influence on glycometabolism in rats with traumatic brain injury （TBI）. Methods： Seventy-eight SD rats were randomly divided into traumatic group （n=36）, RSTN group （n=36） and sham operation group （n=6）. Fluid percussion TBI model was developed in traumatic and RSTN groups and the latter received additional 1 mg RSTN antibody treatment for each rat. At respectively 12 h, 24 h, 72 h, 1 w, 2 w, and 4 w after operation, venous blood was collected and the right hind leg skeletal muscle tissue was sampled. We used real-time PCR to determine mRNA expression of RSTN in skeletal muscles, western blot to determine RSTN protein expression and ELISA to assess serum insulin as well as fasting blood glucose （FBG） levels. Calculation of the quantitative insulin sensitivity check index （Q value） was also conducted. The above mentioned indicators and their correction were statistically analyzed. Results： Compared with sham operation group, the RSTN expression in the skeletal muscle as well as serum insulin and FBG levels revealed significant elevation （P〈0.05）, and reduced Q value （P〈0.05） in traumatic group. Single factor linear correlation analysis showed a significant negative correlation between RSTN expression and Q values （P〈0.001） in traumatic group. Conclusion： The expression of RSTN has been greatly increased in the muscular tissue of TBI rats and it was closely related to the index of glycometabolism. RSTN may play an important role in the process of insulin resistance after TBI.

Glucose metabolic abnormality is associated with defective mineral homeostasis in skeletal disorder mouse model

Bone was reported as a crucial organ for regulating glucose homeostasis. In this study, we found that Phex mutant mice(PUG), a model of human X-linked hypophosphatemic rickets(XLH), displayed metabolic abnormality in addition to abnormal phosphate homeostasis, skeletal deformity and growth retardation. Glucose tolerance was elevated with enhanced insulin sensitivity in PUG, though circulating insulin level decreased. Interestingly, bone mineral density defects and glucose metabolic abnormality were both rescued by adding phosphorus- and calcium-enriched supplements in daily diet. Serum insulin level, glucose tolerance and insulin sensitivity showed no differences between PUG and wild-type mice with rescued osteocalcin(OCN) following treatment. Our study suggested that OCN is a potential mediator between mineral homeostasis and glucose metabolism. This investigation brings a new perspective on glucose metabolism regulation through skeleton triggered mineral homeostasis and provides new clues in clinical therapeutics of potential metabolic disorders in XLH patients.

Effects of electroacupuncture and Chinese kidney-nourishing medicine on polycystic ovary syndrome in obese patients

OBJECTIVE： To explore the effect of electroacu- puncture and Chinese kidney-nourishing medicine on insulin （INS）, adiponectin （APN）, leptin （LEP）, and glucolipid metabolism of obese patients with polycystic ovary syndrome （PCOS）. METHODS： Sixty-seven obese PCOS patients were randomly divided into two groups. Thirty-three pa- tients in the acupuncture-medicine group were treated three times a week with electroacupunc- ture at the Tianshu （ST 25）, Zhongwan （CV ] 2）, Qi- hai （CV 6）, Sanyinjiao （SP 6）, Geshu （BL 17）, and Cil-iao （BL 32） acupoints. They drug, Tiankui capsule, for 3 treatment. Point-taking and also took the Chinese months as a course of treatment in the elec- troacupuncture group of 34 patients was the same as those in the acupuncture-medicine group. We observed and compared the changes in the obesi- ty-related indexes of body weight （BW）, body mass index （BMI）, and waist-hip ratio （WHR）, as well as fasting plasma glucose （FPG）, fasting insulin （FINS）, APN, and LER RESULTS： BW, BMI, WHR, and FINS decreased and insulin sensitivity index （ISI） and APN were higher in the acupuncture-medicine group than in the electroacupuncture group （P〈0.01）. There was no obvious difference in LEP between the two groups （P〉0.05）. CONCLUSION： Acupuncture combined with medi- cine is better than just electroacupuncture for obese PCOS patients by improving obesity-related indexes, insulin sensitivity, and APN level. This indi- cates that acupuncture-medicine therapy is worth clinical popularization.