Electroacupuncture of scalp acupoint alleviates cerebral ischemic inflammatory injury by down-regulating RORγt and promoting balance of IL-17A+Th17/FOXP3+Treg in MCAO rats. / 电头针调控RORγt促进IL-17A+Th17/FOXP3+Treg细胞平衡缓解MCAOå¤§é¼ è„‘ç¼ºè¡€ç‚Žæ€§æŸä¼¤çš„ç ”ç©¶.

OBJECTIVES: To observe the effect of electroacupuncture (EA) of scalp acupoint (Dingnieqian-xiexian, MS6) on expression of retinoid-related orphan receptor γT (ROR γ t), interleukin (IL)-17A, IL-10, transfor-ming growth factor-ß1 (TGF-ß1), IL-6, IL-21, and IL-17A+ Thelper cells(Th) 17 and forkhead transcription factor P3 (FOXP3)+ regulatory T cells (Treg) differentiation of ischemic cortex in ischemic stroke rats, so as to explore its molecular mechanisms underlying relief of inflammatory injury of ischemic stroke. METHODS: A total of 120 male SD rats were randomly assigned to sham operation, model, EA, inhibitor, agonist and EA+agonist groups, with 15 rats in each group. The ischemic stroke model was established by occlusion of the left middle cerebral artery according to Longa's methods. For rats of the EA group and EA+agonist group, EA (2 Hz/100 Hz, 1 mA) was applied to bilateral MS6 for 30 min, once daily for 7 days. Rats of the inhibitor group received intraperitoneal injection of solution of SR1001 (RORγt inhibitor) (2.5 mg/mL, 10 mg/kg), once daily for 7 days. Rats of the agonist and EA+agonist groups received intraperitoneal injection of solution of SR1078 (RORγt agonist) (5 mg/mL, 5 mg/kg) before EA, once daily for 7 days. Rats of the sham operation and model groups were grabbed and fixed in the same way with the other groups. The Zea-longa's score, modified neurological severity score (mNSS) and the neurobehavioral score were assessed before and after the intervention. At the end of experiments, the ischemic cortex tissue was collected. The 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining was used to detect the volume of cerebral infarction. The expression of RORγt mRNA was detected by real-time quantitative PCR；the protein expression levels of RORγt, IL-17A, IL-10 and TGF-ß1 were detected by Western blot；the immunoactivity of IL-6 and IL-21 were detected by immunohistochemistry；the fluorescence areas of IL-17A+Th17 and FOXP3+Treg cells were measured by immunofluorescence and their ratio was calculated in the tissue of ischemic cortex. RESULTS: Relevant to the sham operation group, the model group had a significant increase in the Zea-Longa's score, mNSS score, neurobehavioral score, cerebral infarct volume, expression levels of RORγt mRNA and protein, IL-17A protein, IL-6 and IL-21 immunoactivity, IL-17A+Th17 immunofluorescence intensity, and the ratio of IL-17A+Th17/FOXP3+Treg (P<0.01), and an obvious decrease in the expression levels of TGF-ß1 and IL-10 proteins and FOXP3+Treg immunofluorescence intensity (P<0.01). In contrast to the model group, both EA and inhibitor groups had a significant decrease in the Zea-Longa's score, mNSS score, neurobehavioral score, cerebral infarct volume, expression levels of RORγt mRNA and protein, IL-17A protein, IL-6 and IL-21 immunoactivity, IL-17A+Th17 immunofluorescence intensity, and the ratio of IL-17A+Th17/FOXP3+Treg (P<0.01, P<0.05), and a marked increase in the expression levels of TGF-ß1 and IL-10 proteins and FOXP3+Treg immunofluorescence intensity (P<0.05, P<0.01), while the above indicators of the agonist group were all reversed (P<0.01, P<0.05). Comparison between the agonist and EA+agonist groups showed that the Zea-Longa's score, mNSS score, neurobehavioral score, cerebral infarct volume, expression levels of RORγt mRNA and protein, IL-17A protein, IL-6 and IL-21 immunoactivity, IL-17A+Th17 immunofluorescence intensity, and the ratio of IL-17A+Th17/FOXP3+Treg were significantly lower (P<0.01, P<0.05), and the expression of TGF-ß1 and IL-10 proteins and FOXP3+Treg immunofluorescence intensity were obviously higher (P<0.01, P<0.05) in the EA+agonist group than in the agonist group, suggesting that EA intervention can effectively weaken the effects of RORγt agonist. CONCLUSIONS: EA of scalp acupoint MS6 can effectively improve the neurological function, behavior reaction and reduce cerebral infarct volume in ischemic stroke rats, which may be associated with its functions in down-regulating the expression of RORγt and promoting the balance of IL-17A+Th17/FOXP3+Treg to alleviate inflammatory injury after ischemic stroke.

Suppression of cAMP/PKA/CREB signaling ameliorates retinal injury in diabetic retinopathy.

The blood-retinal barrier (BRB), homeostasis, neuronal integrity, and metabolic processes are all directly influenced by Müller cells, the most important retinal glial cells. We isolated primary Müller cells from Sprague-Dawley (SD) neonatal rats and treated them with glucose at varying doses. CCK-8 was used to quantify cellular viability, and a TUNEL assay was performed to detect cell apoptosis. ELISA, immunofluorescence, and western blotting were used to assess cAMP/PKA/CREB signaling, Kir4.1, AQP4, GFAP, and VEGF levels, respectively. H&E staining was used to examine histopathological alterations in diabetic retinopathy (DR)-affected retinal tissue in rats. As glucose concentration increases, gliosis of Müller cells became apparent, as evidenced by a decline in cell activity, an increase in apoptosis, downregulation of Kir4.1 level, and overexpression of GFAP, AQP4, and VEGF. Treatments with low, intermediate, and high glucose levels led to aberrant activation of cAMP/PKA/CREB signaling. Interestingly, blocking cAMP and PKA reduced high glucose-induced Müller cell damage and gliosis by a significant amount. Further in vivo results suggested that cAMP or PKA inhibition significantly improved edema, bleeding, and retinal disorders. Our findings showed that high glucose exacerbated Müller cell damage and gliosis via a mechanism involving cAMP/PKA/CREB signaling.

lncRNA FGD5-AS1 acts as a ceRNA to regulate lipopolysaccharide-induced injury via the miR-223-3p-3p/GAS5 axis in cardiomyocytes.

Long noncoding RNAs (lncRNAs) are abnormally expressed in numerous diseases, and they are closely associated with cardiac diseases. However, the role of lncRNAs in lipopolysaccharide (LPS)-induced cardiotoxicity as well as the potential mechanism remain largely unclear. In the present study, IncRNA microarray assays were performed to analyze differential lncRNA expression in LPS-treated cardiomyocytes, and lncRNA FGD5-AS1 was one of the downregulated lncRNAs. H9C2 cells were treated with LPS, and the expression of lncRNA FGD5-AS1 was markedly downregulated. LncRNA FGD5 overexpression decreased the LPS-induced cardiomyocyte apoptosis and inflammation. Bioinformatics analysis and a luciferase reporter assay indicated that lncRNA FGD5-AS1 directly binds to miR-223-3p. A miR-222-3p mimic partially reversed the inhibitory effect of lncRNA FGD5-AS1 on the LPS-induced H9C2 cell apoptosis and inflammatory response. Moreover, miR-223-3p directly targeted growth arrest-specific transcript 5 (GAS5). LncRNA FGD5-AS1 regulated LPS-induced H9C2 cell inflammation and apoptosis via the miR-223-3p/GAS5 axis.

Ferroptosis-A Novel Mechanism With Multifaceted Actions on Stroke.

As a neurological disease with high morbidity, disability, and mortality, the pathological mechanism underlying stroke involves complex processes such as neuroinflammation, oxidative stress, apoptosis, autophagy, and excitotoxicity; but the related research on these molecular mechanisms has not been effectively applied in clinical practice. As a form of iron-dependent regulated cell death, ferroptosis was first discovered in the pathological process of cancer, but recent studies have shown that ferroptosis is closely related to the onset and development of stroke. Therefore, a deeper understanding of the relationship between ferroptosis and stroke may lead to more effective treatment strategies. Herein, we reviewed the mechanism(s) underlying the onset of ferroptosis in stroke, the potential role of ferroptosis in stroke, and the crosstalk between ferroptosis and other pathological mechanisms. This will further deepen our understanding of ferroptosis and provide new approaches to the treatment of stroke.

Ethnic differences in the incidence of pterygium in a multi-ethnic Asian population: the Singapore Epidemiology of Eye Diseases Study.

We evaluated the 6-year incidence and risk factors of pterygium in a multi-ethnic Asian population. Participants who attended the baseline visit of the Singapore Epidemiology of Eye Diseases Study (year 2004-2011) and returned six years later, were included in this study. Pterygium was diagnosed based on anterior segment photographs. Incident pterygium was defined as presence of pterygium at 6-year follow-up in either eye, among individuals without pterygium at baseline. Multivariable logistic regression models were used to determine factors associated with incident pterygium, adjusting for baseline age, gender, ethnicity, body mass index, occupation type, educational level, income status, smoking, alcohol consumption, presence of hypertension, diabetes and hyperlipidemia. The overall age-adjusted 6-year incidence of pterygium was 1.2% (95% confidence interval [CI] 1.0-1.6%); with Chinese (1.9%; 95% CI 1.4%-2.5%) having the highest incidence rate followed by Malays (1.4%; 95% CI 0.9%-2.1%) and Indians (0.3%; 95% CI 0.3-0.7%). In multivariable analysis, Chinese (compared with Indians; odds ratio [OR] = 4.21; 95% CI 2.12-9.35) and Malays (OR 3.22; 95% CI 1.52-7.45), male (OR 2.13; 95% CI 1.26-3.63), outdoor occupation (OR 2.33; 95% CI 1.16-4.38), and smoking (OR 0.41; 95% CI 0.16-0.87) were significantly associated with incident pterygium. Findings from this multi-ethnic Asian population provide useful information in identifying at-risk individuals for pterygium.

Melatonin inhibits macrophage infiltration and promotes plaque stabilization by upregulating anti-inflammatory HGF/c-Met system in the atherosclerotic rabbit: USPIO-enhanced MRI assessment.

Macrophage plays critical roles in the pathogenesis of atherosclerosis (AS), and is an attractive target for detecting and treating vulnerable plaque. Our previous study showed that melatonin (MLT) ameliorated AS by suppressing the pro-inflammatory Toll-like receptor 4/nuclear factor kappa B system in high-fat-fed rabbit. However, it is unknown whether the anti-atherosclerotic properties of MLT are associated with the upregulation of anti-inflammatory hepatocyte growth factor (HGF)/mesenchymal-epithelial transition factor (c-Met) system. In present study, we examined whether MLT could inhibit macrophage infiltration and promote plaque stabilization by upregulating HGF/c-Met system with ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance imaging (MRI) assessment in AS rabbit. Rabbits in this study were randomly divided into three groups and treated with a standard diet, high-fat diet, and high-fat diet plus 10 mg/kg/day MLT for 12 weeks, respectively. MLT treatment significantly reversed spotty signal void in 3D-TOF MRI, standard signal intensity reduction in T2WI MRI and aortic luminal area reduction in 2D-TOF MRI of the atherosclerotic abdominal aorta 72 h after USPIO injection. It also decreased serum interleukin-6 (IL-6), intima/media thickness ratio of the abdominal aorta, CD68 and iron-positive areas in the aortic intima, and increased serum IL-10, HGF and c-Met protein expression and the accumulation of vascular smooth muscle cell and collagen fiber in the aortic intima of AS rabbit. Our data demonstrated that MLT significantly decreased plaque macrophage infiltration and promoted plaque stability in AS rabbit assessed by USPIO-enhanced MRI. Remarkably, it was very first revealed that upregulation of anti-inflammatory HGF/c-Met system might contribute to the atheroprotective mechanisms of MLT.

Normative patterns and factors associated with presbyopia progression in a multiethnic Asian population: the Singapore Epidemiology of Eye Diseases Study.

BACKGROUND/AIM: To investigate normative patterns and factors associated with presbyopia progression in a multiethnic Asian population. METHODS: Malay, Indian and Chinese participants aged 40-80 years who had baseline and 6-year follow-up examinations with subjective refraction data were recruited from the Singapore Epidemiology of Eye Diseases Study. Presbyopia progression was defined as an increase in near addition power of ≥+0.50 dioptre (D) from baseline to follow-up visit. Modified Poisson regression analyses were used to determine baseline factors associated with presbyopia progression. RESULTS: From the eligible 3974 eyes, 2608 eyes were included for final analysis after excluding eyes with a history of cataract surgery (929 eyes) and best-corrected distance visual acuity worse than 20/40 (342 eyes). Overall the mean near addition power change over 6 years was +0.25 D; Malays showed greater change (+0.37 D) compared with Indians (+0.23 D) and Chinese (+0.16 D). After adjusting for baseline age, gender, body mass index, hypertension, cataract, refractive error and daily hours of reading and writing, Malays were more likely to have presbyopia progression compared with Chinese (RR (relative risk)=1.67; 95% CI 1.43 to 1.95; p<0.001) and Indians (RR=1.45; 95% CI 1.25 to 1.68; p<0.001). Individuals aged 60-69 years (RR=0.77; p=0.006) and ≥70 years (RR=0.51; p<0.001) were less likely to progress in presbyopia compared with those aged 40-49. CONCLUSION: In this Asian population, the near addition power change over 6 years was lower than the current near addition prescription guidelines (+0.25 D vs +0.60 D). Our findings may help update near addition prescription guidelines that can be more tailored to Asians.

Association between polymorphism rs11200638 in the HTRA1 gene and the response to anti-VEGF treatment of exudative AMD: a meta-analysis.

BACKGROUND: Anti-angiogenesis treatments are the most commonly used treatments for the vision loss caused by exudative age-related macular degeneration (AMD), in which the anti-vascular endothelial growth factor (VEGF) drugs with ranibizumab and bevacizumab are current standard treatments. However, the outcome of anti-VEGF therapeutics is not uniform in all patients. METHODS: We performed a literature-based meta-analysis including, five published studies relevant to HTRA1 and response to anti-VEGF treatment (bevacizumab or ranibizumab). Summary odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using fixed- and random-effects models. Sensitivity analysis and meta-regression were also performed. Q-statistic test and Egger's test was used to evaluate heterogeneity and publication bias respectively. RESULTS: Overall, no association between the rs11200638 polymorphism in HTRA1 gene and the anti-VEGF treatment response was found in the genotype GG versus AA (OR = 1.06; 95% CI: 0.77 to 1.48; P = 0.98), genotype GA versus AA (OR = 1.11; 95% CI: 0.83 to 1.47; P = 0.93), genotype GG + GA versus AA (OR = 1.22; 95% CI: 0.94 to 1.57; P = 0.09), and allele G versus A (OR = 0.92; 95% CI: 0.78 to 1.08; P = 0.14). In the subgroup analysis by ethnicity Caucasian population, and a significant association was still not observed in all genetic models. Sensitivity analysis indicated the robustness of our findings, and no publication bias was observed in our meta-analysis. CONCLUSIONS: This study shows that there was no association between the polymorphism rs11200638 in HTRA1 gene and response to anti-VEGF treatment of exudative AMD. However, more studies are needed to further prove the conclusion of present study, especially well-designed and high quality randomised controlled trials or intervention studies.

Internal limiting membrane peeling or not: a systematic review and meta-analysis of idiopathic macular pucker surgery.

PURPOSE: To determine whether internal limiting membrane (ILM) peeling improves anatomical and functional outcomes in idiopathic macular pucker (IMP)/epiretinal membrane (ERM) surgery in this systematic review and meta-analysis. METHODS: We searched the PubMed, Medline, Web of Science, Cochrane, Ovid MEDLINE, ClinicalTrials.gov and CNKI databases for studies published before 15 September 2016. The eligibility criteria included studies comparing ILM peeling versus no-peeling for IMP surgery. RESULTS: Thirteen articles (10 retrospective cohort studies, 1 prospective cohort study and 2 randomised controlled trials (RCTs)) were included in the review. Primary outcomes: no differences were observed in the best-corrected visual acuity (BCVA) or central macular thickness (CMT) at 12 months; however, lower ERM recurrence (OR, 0.13; 95% CI 0.04 to 0.41; p=0.0004) and reoperation rates (OR, 0.10; 95% CI 0.02 to 0.49; p=0.004) that favoured ILM peeling were observed at the final follow-up. SECONDARY OUTCOMES: no difference was observed in BCVA at 3, 6 months, the final follow-up or in CMT at 3, 6 months, the final follow-up. Significantly increased CMT, which favoured ILM peeling, was observed at the final follow-up (p=0.002) in the RCTs. CONCLUSIONS: ILM peeling yielded greater anatomical success, but no improvement in functional outcomes as the treatment of choice for patients undergoing IMP surgery.

[Preparation, characterization of paclitaxel-loaded Pluronic P105 polymeric micelles and in vitro reversal of multidrug resistant tumor].

Drug delivery system (DDS) is a novel approach to overcome multidrug resistance (MDR) in tumors nowadays. This work was designed to investigate a new micellar delivery system for in vitro reversal of resistant ovarian tumor cells, based on a nonionic triblock copolymer Pluronic P105 and paclitaxel (PTX). The PTX-loaded polymeric micelles (P105/PTX) were prepared by thin film-hydration methods. Based on the results of single factor experiments, the P105/PTX micelle formulation was optimized by employing the central composite design-response surface methodology. The physico-chemical properties of the P105/PTX micelles were characterized, including micelle size, drug loading coefficient, in vitro release behavior, etc. The cytotoxicity of the P105/PTX micelles was assessed against human ovarian tumor cell line, SKOV-3/PTX, by a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT) assay. In order to understand the possible mechanism of Pluronic effects in resistant tumor cells, cellular uptake study of micellar PTX or Rhodamine-123 (R-123) was also carried out. The results showed that the micelle size was about 24 nm with drug loading coefficient of 1.1% and PTX concentration of 700 microg x mL(-1). The cumulative release amount of PTX from the P105/PTX micelles was only 45.4% in 6 h (P < 0.05) and 79.6% in 24 h, whereas Taxol injection in 6 h released 95.2% PTX. The IC50 values of the P105/PTX micelles and Taxol injection against SKOV-3/PTX were 1.14 and 5.11 microg x mL(-1), and resistance reversion index (RRI) was 9.65 and 2.15, respectively. The micellar PTX or R-123 exhibited a significant increase in cellular uptake in resistant SKOV-3/PTX cells compared with free PTX or R-123. These results indicated that PTX could effectively be solubilized by Pluronic P105 block copolymers via thin film-hydration process and formulation optimization, producing nano-scale polymeric micelles with sustained release property in vitro. The P105/PTX micelles were effectively able to reverse resistance to PTX in SKOV-3/PTX tumor cells compared with Taxol injection or free PTX solution, and the enhanced cytotoxicity in the resistant SKOV-3/PTX cell was related to the improved cellular uptake of PTX by Pluronic P105 copolymers.

[Effect of carbon dioxide pneumoperitoneum-laparoscopic surgery on tumor seeding and metastases in endometrial cancer].

OBJECTIVE: To explore the influence of carbon dioxide pneumoperitoneum-laparoscopic surgery on tumor cell seeding and metastases in endometrial cancer. METHODS: Twenty patients with endometrial cancer who underwent laparoscopic surgery and 10 patients with endometrial cancer who underwent laparotomic surgery were enrolled. Each patient was in preoperative clinical StageIand the uterus size in each patient was less than 12 weeks of pregnancy. Carbon dioxide pneumoperitoneum was established and maintained with CO2 insufflation at 4 approximately 6 L/min and intraperitoneal pressure of 13 mmHg with an automatic pneumoperitoneum machine. Cytologic examination of peritoneal fluid(at the beginning and end of the operation), CO2 filtrated gas and the lavage fluid of instruments during the laparoscopic surgery were performed. The protein expressions of E-cadherin,beta-catenin,P-selectin,matrix metalloproteinase-2(MMP-2),vascular endothelial growth factor (VEGF),and CD44v6 in tumor tissues before and after the operation were detected by DAKO Envision. RESULTS: There were no case of positive washing cytology in the peritoneal fluid,CO2 filtrated gas, and the lavage fluid of instruments during the laparoscopic surgery. The expressions of E-cadherin and beta-catenin proteins were obviously abnormal in endometrial cancer. The abnormal expressions of E-cadherin and beta-catenin protein between the pre- and post-operations were not significantly different in both the laparoscopic group and the laparotomic group(P>0.05).The changes of abnormal expressions of E-cadherin and beta-catenin protein were no statistical difference between the two groups(P>0.05). The positive protein expressions of P-selectin,MMP-2,VEGF,and CD44v6 were not significantly different between the pre- and post-operations both in the laparoscopic group and the laparotomic group(P>0.05),and there was also no significant difference between the laparoscopic group and the laparotomic group(P>0.05).The follow-up period in the laparoscopic group was 7 approximately 19 (14.25+/-3.65) months and 7 approximately 19 (13.10+/-4.23) months in the laparotomic group. One patient got infection in the urinary system in the laparoscopic group and one patient had lower extremity venous thrombosis in the laparoscopic group.No recurrence was detected in both groups. CONCLUSION: Laparoscopic surgery for endometrial cancer has no effect on protein expressions of E-cadherin,beta-catenin,P-selectin,MMP-2,VEGF,and CD44v6 in tumor tissues. No evidence has been found that CO2 pneumoperitoneum-laparoscopic surgery may favor endometrial cancer cell seeding and metastases.

Biodistribution in mice and severity of damage in rat lungs following pulmonary delivery of 9-nitrocamptothecin liposomes.

This study was designed to investigate in vitro release, in vivo tissue distribution and the damage to the lungs of 9-nitrocamptothecin (9-NC) liposomes. In vitro release of 9-NC from liposomes was carried out in phosphate buffer saline solution (PBS) pH 7.4. The tissue distribution of 9-NC liposomes and 9-NC solution was determined after pulmonary delivery to mice. The tissue distribution of 9-NC liposomes after intravenous administration was also studied. The changes of pulmonary edema index and histology of lungs in rats were investigated to evaluate the severity of the damage after pulmonary delivery. The results showed that 9-NC was continuously released from liposomes in PBS pH 7.4 for 24h at 37 degrees C. After pulmonary delivery, the mean residence time (MRT) of 9-NC liposomes in the lungs was 3.4 times as long as that of 9-NC solution and the total AUC0-t of all tissues in mice of the liposomes was 2.2-fold higher than that of the solution, indicating that the liposomes had sustained-release characteristics. Following intravenous administration and pulmonary delivery, the targeting efficiency (Te) to the lung of 9-NC liposomes was 0.14 and 2.02, respectively, which showed that intratracheal instillation can deliver the drug mainly to the lung and decrease the accumulation of the drug in other tissues at different concentrations. The pulmonary edema index and the histological changes of the lungs in 9-NC liposome group were significantly different from those in 9-NC solution group. The lung damage by liposomes was less severe than that by solution. Pulmonary delivery of 9-NC liposomes could directly deliver the drug to the lung and make the drug accumulate in the lung with sustained-release characteristics, changing the disposition behavior in vivo, decreasing the toxic and side effects on other tissues and reduce the severity of damage to lungs following intratracheal instillation.

[Screening of Panax notoginsenoside water in oil microemulsion formulations and their evaluation in vitro and in vivo].

Water in oil (W/O) microemulsion formulation was developed to enhance intestinal absorption of ginsenoside Rb1 (Rb1) of panax notoginseng (PNS). Effects of W/O microemulsions on pharmacokinetics after intraduodenal administration, membrane fluidity and membrane transport of Rb, were studied in rats, liposomes and parallel artificial membrane permeability assay (PAMPA), respectively. Soybean phospholipids/ethanol (SP/EtOH) was selected as surfactant/cosurfactant, together with PNS 400 mg x mL(-1) solution and various kinds of oils, to prepare 11 W/O microemulsions. Most of the microemulsions can enhance Rb1 intestinal absorption significantly. Besides surfactant/cosurfactant, oil also had an effect on the enhanced absorption and the order of enhancement was as follows: glyceryl laurate approximately = isopropyl myristate > isopropyl palmitate > 2-ethylhexanol palmitate. The effection of absorption enhancement by the long chain glyceride ( > C14) is lower than that by the medium chain glyceride (C8 - C14). Most of W/O microemulsions were found to enhance the membrane fluidity of liposomes to different extents. In PAMPA analysis, efficient permeability coefficient (Pe) of diluted-microemulsion (D-ME) is mostly higher than that of PNS solution, which indicated the components of microemulision can facilitate the membrane permeability of the drug. Meanwhile, linearity correlation between Pe and ratio of relative bioavailability (Fr) was acquired for undiluted microemulison (ME). Therefore, W/O microemulsions can enhance intestinal absorption of Rbr, and this effect may be attiributed to its enhancement on membrane fluidity to a certain degree. PAMPA analysis could be brought into not only the investigation of membrane transport of crude drug, but also conditioned preformulation research (e.g. absorption enhancer etc.).

Pharmacokinetics and biodistribution of polymeric micelles of paclitaxel with Pluronic P123.

AIM: To investigate the preparation, in vitro release, in vivo pharmacokinetics and tissue distribution of a novel polymeric micellar formulation of paclitaxel (PTX) with Pluronic P123. METHODS: The polymeric micelles of paclitaxel with Pluronic P123 were prepared by a solid dispersion method. The characteristics of micelles including particle size distribution, morphology and in vitro release of PTX from micelles were carried out. PTX-loaded micellar solutions were administered through the tail vein to healthy Sprague-Dawley rats and Kunming strain mice to assess the pharmacokinetics and tissue distribution of PTX, respectively. Taxol, the commercially available intravenous formulation of PTX, was also administered as control. RESULTS: By using a dynamic light scattering sizer and a transmission electron microscopy, it was shown that the PTX-loaded micelles had a mean size of approximately 25 nm with narrow size distribution and a spherical shape. PTX was continuously released from Pluronic P123 micelles in release medium containing 1 mol/L sodium salicylate for 24 h at 37 centigrade degree. In the pharmacokinetic assessment, t(1/2beta) and AUC of micelle formulation were 2.3 and 2.9-fold higher than that of Taxol injection. And the PTX-loaded micelles increased the uptake of PTX in the plasma, ovary and uterus, lung, and kidney, but decreased uptake in the liver and brain in the biodistribution study. CONCLUSION: Polymeric micelles using Pluronic P123 can effectively solubilize PTX, prolong blood circulation time and modify the biodistribution of PTX.

[Effect of progesterone on the secretion of matrix metalloproteinase-2 and matrix metalloproteinase-9 in human ectopic endometrial stromal cells].

OBJECTIVE: To determine the effect of progesterone on the secretion of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in ectopic endometrial stromal cells. METHODS: Ectopic endometrial stromal cells were obtained from 17 patients with endometriosis. Endometrial stromal cells were obtained from 12 patients with endometriosis and 14 cases of controls. Ectopic endometrial stromal cells of 15 cases were treated with progesterone. Culture supernatants of these stromal cells were analyzed for MMP-2 and MMP-9 by zymography. RESULTS: Endometriotic stromal cells released significantly higher levels of MMP-2 and MMP-9 than endometrial stromal cells from women with and without endometriosis. Progesterone at 10(-9) mol/L caused endometriotic stromal cells a significant reduction MMP-2 and MMP-9 levels. When progesterone concentration was increased from 10(-9) mol/L to 10(-7) mol/L, the release of MMP-9 was almost completely inhibited, wherease that of MMP-2 was not completely inhibited. CONCLUSION: Progesterone may inhibit the secretion of MMP-2 and MMP-9 in ectopic endometrial stromal cells, especially MMP-9.

[Expression of cylooxygenase-2 in endometriosis].

OBJECTIVE: To investigate the expression of cyclooxygense-2 (COX-2) in eutopic and ectopic endometrium in ovarian endometriosis. METHODS: Thirty patients with ovrian endometriosis, 10 with ovarian chocolate cysts and 27 normal controls were enrolled it determine the expression of COX-2 immunohistochemically in eutopic or ectopic endometrium or healthy endometrial tissues. RESULTS: The immunoreactivities of COX-2 were found in epithelial cells and stromal cells in eutopic endometrium. The expression of COX-2 in the epithelial cells in the secretory phase was higher than that in the proliferative phase in the control group and ovarian endometriosis group (P <0. 05). But the expression of COX-2 in stromal cells in the control group and ovarian endometriosis group showed no cyclic changes throughout the menstrual cycle (P > 0. 05). The expression of COX-2 in eutopic and ectopic endometrium in the ovarian endometriosis group was higher than that in the control group (P <0. 05) , hut we did not find significant difference between the eutopic and ectopic endometrium in the ovarian endometriosis group (P > 0. 05). CONCLUSION: The increased COX-2 expression in eutopic and ectopic endometrium in ovarian endometriosis may he related to its pathology.

[9-nitrocamptothecin nanostructured lipid carrier system: in vitro releasing characteristics, uptake by cells, and tissue distribution in vivo].

AIM: To study the release and cell uptake characteristics of 9-nitrocamptothecin (9-NC) nanostructured lipid carrier system (NLC) in vitro and its tissue distribution characteristics in vivo. METHODS: Mouse peritoneal macrophages were used to investigate the uptake of nanoparticles by cells in vitro. The tissue distribution of 9-nitrocamptothecin solution and stealth nanostructured lipid carrier system (S-NLC) was determined after intravenous administration to mice at a single dose of 1.5 mg kg(-1). The release and crystalloid characteristics were also investigated. RESULTS: X-ray diffraction spectrum showed that 9-NC probably was amorphous in S-NLC. The liquid lipid did not change the characteristics of the solid matrix in nanoparticles. The in vitro release and cell uptake characteristics of stealth and non-stealth 9-NC-NLC were investigated, separately. The results showed that the stealth 9-NC-NLC had sustained-release characteristics and could resist the absorption effect of the additional plasmas to a certain extent. In addition, the cell uptake percentage of stealth 9-NC-NLC was much lower than that of the non-stealth ones. The tissues distribution results showed that 9-NC in the S-NLC was mainly found in the lung, liver, pancreas and ovary/uterus, while the quantity of 9-NC was much lower in heart and kidney. The AUQ(0-t), of S-NLC in blood, ovary/uterus, pancreas, liver and lung were higher than that of 9-nitrocamptothecin solution. The weight-average drug targeting efficiency (Te*) of S-NLC in liver and lung were significantly higher than that of 9-nitrocamptothecin solution. The mean residence times (MRT) of S-NLC was 44 h, while that of 9-nitrocamptothecin solution was 8 h. Therefore, S-NLC showed obvious targeting effects on liver and lung. CONCLUSION: S-NLC with PEG flexible chains has sustained-release characteristics and can prolong its circulation in blood and have good targeting efficiency on liver and lung.

[The in vitro kinetics of uptake, transport and efflux of 9-nitrocamptothecin in Caco-2 cell model].

AIM: To study the kinetics of uptake, transepithelial transport and efflux of 9-nitrocamptothecin (9-NC). METHODS: A human intestinal epithelial cell model Caco-2 cell in vitro cultured had been applied to study the kinetics of uptake, transport and efflux kinetics of 9-NC at small intestine. The effects of time, pH, temperature and P-glycoprotein inhibitors on the uptake of 9-NC were investigated. The determination of 9-NC was performed by HPLC. RESULTS: The uptake and absorption of 9-NC were passive diffusion as the dominating process. The uptake of 9-NC is positively correlated to uptake time, and negatively correlated to pH and temperature. The inhibitors, cyclosporine A and verapamil, significantly enhanced the uptake amount of 9-NC (P < 0.05). P(app) of Basolateral to Apical was much more than that of Apical to Basolateral (2.6-6.9 fold). The efflux of 9-NC was fitted to apparent two-order process. The m0 [(148.0 +/- 2.2) pmol x cm(-2)] and the efflux rate (41.1 pmol x cm2 min(-1)) on Apical side were higher than the m0 [(121 +/- 7) pmol x cm(-2)] (P < 0.05) and the efflux rate (29.2 pmol x cm2 x min(-1)) on Basolateral side (P < 0.01). CONCLUSION: The uptake and absorption of 9-NC were passive diffusion as the dominating process. P-glycoprotein had strong efflux effects on the uptake and transepithelial transport of 9-NC.

[Expression of vascular endothelial growth factor in the peritoneal fluid and eutopic endometrium of patients with endometriosis].

OBJECTIVE: To investigate the significance of vascular endothelial growth factor (VEGF) on the etiology of endometriosis. METHODS: The level of VEGF in the peritoneal fluid from women with endometriosis (n = 19) and non-endometriosis (n = 21) was studied by ELISA. The expression of VEGF of eutopic endometrium collected from women with endometriosis (n = 15) and non-endometriosis (n = 12) was determined by relative quantity RT-PCR. RESULTS: The level of VEGF in the peritoneal fluid in the endometriosis was higher than that of the controls (P < 0.05). Cyclic variation in VEGF concentration was seen in the peritoneal fluid from patients with endometriosis, and the VEGF concentration in the proliferative phase was significantly higher than that in the secretory phase (P < 0.05). No cyclic variation in VEGF was seen in the control group (P > 0.05). The expression of VEGF in the eutopic endometrium with endometriosis was significantly higher than that of the controls (P < 0.001). There was no significant difference between the proliferative and secretory phase in endometriosis (P > 0.05). CONCLUSION: The increase of VEGF in the peritoneal fluid and eutopic endometrium with endometriosis may be important to the occurrence and development of endometriosis.