Objective To investigate the expression pattern of carbonic anhydrase 1 (CA1) in different clinical stages of color ectal cancers and the correlation between CA1 expression and pathologic characteristics of colorectal cancer.Methods The expression of CA1 in genetic level was detected by real-time quantitative polymerase chain reaction (PCR) and the expression of CA1 in protein level was detected by Western blot and immunohistochemical staining.Results The expression pattern of CA1 in the protein level was very similar to the genetic level.It had a very low expression level in carcinoma tissue,especially at stages Ⅲ and Ⅳ (P ＜0.01),CA1 had significantly lower expression level in the colorectal patients with metastasis (P ＜0.01).CA1 was gradually decreased from well-differentiation tissue to poorly-differentiation tissue with a significant difference between well-differentiated adenocarcinoma and poorly-differentiated adenocarcinoma (P ＜ 0.01).Kaplan-Meier survival analysis showed that the expression of CA1 was extremely related to colorectal carcinoma (CRC) prognosis.The 5-year survival rate was only 8％ in the CA1 negative expression group,and the 5-year survival rate of the patients with CA1 positive expression was 92.86％ (P ＜ 0.01).Conclusions CA1 had the similar expression patterns at genetic and protein levels.In the colorectal cancer tissue,the expression of CA1 was downregulated and even missing.CA1 had significantly lower expression level in the advanced colorectal cancers with metastasis and poorly differentiated colorectal cancer tissues.The patients with lower CA1 expression level had the worse prognosis.

Objective To investigate the effect of α-crystallin on the proliferation of rat retinal microglia after optic nerve injury. Methods The effect of α-crystallin on number and proliferation of microglia were analyzed by MTT assay.After the rat model with optic nerve injury was established,α-crystallin was iniected into vitreous cavity and the microglia cell number were counted and compared by retinal fiat counting and immunofluorescence labeling in different groups. Results The proliferation and activation of microglia cells could be stimulated by LPS at 10-6g/L to 10-2g/L.α-crystallin at 10-4g/L and 10-6g/L could inhibit proliferation and activation of microglia cells.Compared to BSA iniection group,α-crystallin could inhibit more significantly the number of microglia cells 1-3 weeks after injury (P＜0.05). Conclusions α-crystallin can inhibit proliferation and activation of retinal microglia and alleviate overphagocytosis and secondary damage of retinal microglia to retinal ganglion cells(RGCs),which may be another mechanism that α-crystallin contributes to indirect protection of RGCs.

ObjectiveTo study the state of oxidative injury induced by sodium arsenite(NaAsO2) in SV-40-immortalized normal uroepithelial (SV-HUC-1 ) cells.Methods SV-HUC-1 cells were exposed to different concentrations of NaAsO2[0(control),1,2,4,8,10 μmol/L] for 24 h,intracellular reactive oxygen species (ROS) was determined by flow cytometry,and the content ofintracellular nitrotyrosine(NT) and the 8-Hydroxydeoxyguanosine (8-OHdG) levels of cell culture medium were detected by enzyme linked immunosorbent assay (ELISA).Results After 24 h treatment,ROS levels(81.76 ± 4.91,95.23 ± 2.17,126.61 ± 17.95,126.74 ± 27.77,114.18 ± 9.65) of SV-HUC-1 cells in the 1,2,4,8,10 μmol/L NaAsO2 exposure groups were significantly higher than those of the control group (69.84 ± 1.28,P ＜ 0.05 or ＜ 0.01 ),ROS levels and exposure dose were positively correlated significantly(r =0.818,P＜ 0.01); the content of NT in the 10 μmol/L NaAsO2 exposure group[(919.66 ± 206.33) μg/L] was significantly higher than that in the control group[ (238.19 ± 38.28)μg/L,P ＜ 0.01 ],NT content and dye concentrations of arsenic also had dose-response relationship (r =0.617,P ＜ 0.01); after 24 h the cells were treated with arsenic,no significant difference of 8-OHdG content in the culture medium was observed(F =2.127,P ＞ 0.05 ).ConclusionNaAsO2 can cause SV-HUC-1 cell oxidative damage.

Objective To analyze the risk factors and correlation between clinical indicators and the four main pathological lesions of IgA ne?phropathy in the Oxford classification:mesangial hypercellularity(M0/1),endocapillary proliferation(E0/1),segmental sclerosis or adhesion(S0/1), and tubular atrophy/interstitial fibrosis(T0/1/2). Methods Clinical and pathological data were collected from 514 patients with biopsy?proven IgA nephropathy admitted in our hospital from February 17,2006 to October 11,2011. These patients were all above 18 years old. Cases with sec?ondary causes of mesangial IgA deposition were excluded,such as Henoch?chonlein purpura,ankylosing spondylitis and psoriasis et al. The inde?pendent risk factors affecting the pathological classification were analyzed by Spearman rank correlation analysis and two?category and multi?classi?fication logistic regression using SPSS 17.0 statistical software. Results In 514 IgAN patients,the ratio of males to females was 1.06:1. The aver?age age was 35.70±11.99 years,and the average disease duration was 18.31±30.42 months. M0E0S0T0 was the major pathologic classification of isolated hematuria. Chronic kidney disease(CKD)stage,24 hours proteinuria,albuminuria,urine transferrin and IgG levels were positively corre? lated with M lesion;serum albumin,C3 and PLT showed a negative correlation with M lesion. Twenty four hours proteinuria and blood platelet count were the independent risk factors for M lesion. As shown by stratified analysis ,the proportion of M1 in cases with 24 hours proteinuria≥3.5 g/d is much higher than that in cases with non?nephrotic range proteinuria. Age,systolic blood pressure,uRBC,24 hours proteinuria,albuminuria urine transferrin and IgG levels were positively correlated with E lesion,Duration,serum albumin showed a negative correlation with E lesion. Age and duration of nephritis were independent risk factors for E lesion. 73.3%of patients that above 60 years old showed endothelial proliferation. CKD stage,24 hours proteinuria were positively correlated with S lesion. Age,CKD stage,systolic blood pressure,diastolic blood pressure,C4,TC, LDL?C,CRP,Fib,UA,Cys?C and 24 hours proteinuria,urineβ2?microglobulin,albumin,transferrin and IgG levels were positively associated with T lesion;hemoglobin,serum albumin,serum IgG showed a negative correlation with T lesion. Infection history,high CRP levels,DBP more than 90 mmHg,hypoalbuminemia,high low density lipoproteinemia,and anemia were independent risk factors for T lesion. Conclusion Twenty four hours proteinuria,blood platelet count,age,duration of nephritis,hypoalbuminemia,anemia,hyperlipidemia,DBP≥90 mmHg and high CRP lev?els were risk factors for the Oxford classification of IgA nephropathy. Renal biopsy should be carried out in time to make clear the pathological clas?sification and individual treatment,so as to improve the prognosis.

OBJECTIVETo investigate the protective effect of Shenfu Injection against ischemia-reperfusion (I/R) injury due to pancreas transplantation in rats, and explore its possible mechanism.

METHODSSix normal SD rats with sham operation were taken as the normal control group, 24 steptozozin-induced diabetic SD rats were randomly divided into 4 groups, with 6 in each group. Except I/R group, the rats in the other groups were intravenous injected with Shenfu Injection (SF,10 mg/kg), Hongshen Injection (HS, 9 mg/kg) and Fuzi Injection (FZ 1 mg/kg) respectively at the day and 30 minutes before pancreas transplantation performed in the SF group, HS group and FZ group, respectively. At the same time, rats in the normal control group and in the I/R group were intravenously injected the same volume of normal saline. The blood glucose was detected before and after reperfusion, and 2 hours later after reperfusion, the contents of serum nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), the concentrations of malondialdehyde (MDA) , superoxide dismutase (SOD) , and myeloperoxidase (MPO) in the transplanted pancreas tissues were detected. The cell apoptosis of the transplanted pancreas tissue was determined by TUNEL, and the bcl-2 and Bax protein expression was determined by Western blot.

RESULTSAfter reperfusion, the levels of blood glucose and TNF-alpha decreased and the concentration of NO increased in the SF group, HS group and FZ group, compared with those in the I/R group. The activity of SOD, bcl-2 expression and the ratio of bcl-2 and Bax were higher, while the content of MDA, the activity of MPO, apoptotic indexes, and Bax expression were lower in the SF group, HS group and FZ group than those in the I/R group.

CONCLUSIONShenfu Injection can protect L/R injury due to pancreas transplantation in rats, the possible mechanism may be related to promoting activity of SOD, increasing synthesis of endogenous NO, decreasing the excretion of TNF-alpha, alleviating conglutination and aggregation of polymorphonuclear neutrophils (PMNs) in pancreas, as well as up-regulating Bcl-2 gene expression and down-regulating the Bax gene expression.

Animals ; Cell Aggregation ; drug effects ; Diabetes Mellitus, Experimental ; enzymology ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Injections ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Pancreas ; drug effects ; metabolism ; Pancreas Transplantation ; adverse effects ; Protective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection

LC-MS/MS method was used to simultaneously determine anti-oxidative active catechins EGCG, ECG, EGC and EC in plasma of rats treated with tea polyphenols (TP). The integrated plasma concentration (C') of TP was calculated by means of self-defined weighing coefficient based on percent AUC of individual components, thereby assessing integrated pharmacokinetic (PK) parameters of TP via log C'-T curve. The anti-free radical effects of TP were estimated using inhibitory rate of drug-containing serum collected at different times from rats against in vitro lipid peroxidation of mouse liver homogenate. The obtained E-T curves were used to calculate anti-free radical pharmacodynamic (PD) parameters of TP. E-logC and E-log C' plots and linear regression were carried out in order to obtain the correlation coefficient (R2). The results indicated that the log C'-T curves of TP, which could be best described by three-compartment model, corresponded to elimination rule of iv administration of drugs. The integrated PK parameters showed that TP was distributed in body rapidly and widely, and eliminated from deep compartment slowly. From comparison of R2 values and consistence of C'-T course and E-T course, it was evident that TP integrated PK behaviors correlated much better with its PD behaviors than individual active components, and thus demonstrated that integrated PK parameters could characterize to maximal extent holistic disposition of Chinese herbal drugs and reflect residence properties of holistic effective substances in biological body.

Objective To explore the effects of combination of traditional Chinese medicine (TCM)and western medicine on a variety of allergen induced severe perennial seasonal allergic rhinitis patients who were treated invalidly with antihistamines and intranasal flixonase for more than two years.Methods 16 patients with moderate to severe allergic rhinitis who were uncontrolled with antihistamines and intranasal flixonase for one week were treated by combination with TCM of Xin-Yi-Qing-fei San or Gyokubeifu-san mixed formula for two weeks.Nasal symptoms,its effects on the quality of daily life,and telephone follow-up half a year later were evaluated.Mini Rhinoconjunctivitis Quality of Life Questionnaire (Mini-RQLQ) and Nocturnal Rhinoconjunctivitis Quality of Life Questionnaire (NRQLQ) were used to evaluate the quality of life.Results After 2 weeks combination-treatment,the nasal symptoms of congestion,rhinorrhea,sneezing,nasal itching,and nasal obstruction were significantly improved compared with combination-before [(52.0± 11.1)％,(69.1±5.3)％,(68.3±12.3)％,(55.5±10.0)％to (10.9±2.4)％,(8.9±3.2)％,(12.5±4.3)％,(13.2±5.3)％,respectively].The before and after treatment of nasal function of nasal minimum cross-sectional area (MCA),nasal cavity volume (V5) and nasal airway resistance (R75 and R150) respectively were (0.58±0.12)cm2,(0.38 ± 0.23) Pa/cm3 · s-1,(0.44 ± 0.32) Pa/cm3 · s-1,(3.50 ± 2.33) cm2 to (0.48 ± 0.23) cm2,(0.31 ±0.33) Pa/cm3 · s-1,(0.31 ±0.37)Pa/cm3 · s-1,(2.24± 1.03) cm2 (P＜0.01).In addition,findings of patients with nasal symptoms by half year follow-up were significantly better than those of before.Conclusion The interventionof TCM can effectively control the perennial and seasonal severe symptoms of patients with AR,and improve the quality of life of patients.

OBJECTIVE: To investigate the effect of capsaicin on the binding rates of simvastatin with rats and healthy subjects plasma. METHODS: The plasma protein binding rates of simvastatin in the presence or absence of capsaicin were determined by ultrafiltration combined with liquid chromatography-mass spectrometry (LC-MS/MS). RESULTS: The binding rates of simvastatin (0.2, 0.5, 1 μg·mL-1) were (97.73±0.24)%, (96.75±2.58)%, and (96.23±0.81)% in rat plasma and (97.05±1.74)%, (97.68±0.65)%, and (97.20±1.00)% in human plasma, respectively. In the presence of capsaicin (1 μg·mL-1), the plasma protein binding rates of simvastatin decreased to (93.97±0.96)%, (92.76±1.89)%, (92.05±0.58)% and (91.71±2.51)%, (91.73±1.88)%, (91.41±2.48)%, while the unbound drug concentration increased by 1.66, 1.23, 1.11 fold and 1.81, 2.56, 2.07 fold, respectively. CONCLUSION: Capsaicin could significantly decrease the plasma protein binding rate of simvastatin and increase its free drug concentration (P<0.01). When the clinical use of simvastatin, attention should be paid to the effect of drugs/ food containing capsaicin on its plasma protein binding rate.

BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.