Objective To investigate the cytotoxicity and radiosensitization of 5-nitroindazole-3-formyliminodiacetic acid on HeLa cells.Methods HeLa cells in exponential growth phase were incubated in culture media with different doses and the survival rate was determined by MTT assay.The survival rate of cells receiving radiation combined with different doses of medicine was compared with that of the control.Results The cytotoxicity of S-nitroindazole-3-formyliminodiacetic acid on HeLa cells was very low.The drug had hypoxia radiosensitizing effect on HeLa cells.At doses of 0,6,12,24,48 and 96 μg/ml under hypoxia ,the survival rate were 0.91 ,0.87,0.84,0.81 ,0.76 and 0.60,respectively.At the dosage of 48 and 96 μg/ml,the survival rate were 0.85 and 0.73 under oxygenous).Conclusions 5-Nitroindazole-3-formyliminodiacetic acid has low cytotoxicity and rediosensitizing effect on HeLa cells.

Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.

Objectives To investigate the changes of cystatin C in the serum of patients with acute kidney injury ( AKI) or end-stage renal disease ( ESRD), and study its significance in the early diagnosis of AKI and its correlation with the degree of renal function injury. Method The cases in Xiangya hospital were enrolled in this study according to the RIFLE criteria, including 20 cases of slight acute kidney injury, 30 cases of medium-severe acute kidney injury, 48 cases of victims of the 5. 12 wenchuan earthquake, 32 cases of end-stage renal disease and 20 healthy patients. The microparticle-enhanced immunoturbidimetry method was used to detect serum cystatin C, and the colorimetric method was used to detect urine N-acetyl-beta-D-glucosaminidase (NAGase). The enzymic method was used to detect serum creatinine. The correlation between serum cystatin C and serum creatinine was analyzed, and the sensitivity and specificity of serum cystatin C were evaluated with the receiver operator characteristic curve (ROC curve). Results Compared with healthy control group, the serum cystatin C increased obviously in acute kidney injury group and ESRD group( P <0. 05 and P <0. 01). The serum cystatin C was positively correlated with serum creati-nine( P <0.01). The serum cystatin C in the 5. 12 wenchuan earthquake injured group was also higher than that in healthy control group ( P <0.05), an the serum cystatin C had an AUC - ROC of 0.931 ( P <0. 01). Conclusion Compared to the conventional biomarkers, the earlier emergence of serum cystatin C can contribute better to early clinical diagnosis of AKI. The serum cystatin C is positively correlated with renal function, and reflect changes in renal function accurately.

Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.

OBJECTIVE:To compare the performance of several pattern recognition methods in the identification of volatile oils of traditional Chinese medicine(TCM)by infrared spectroscopy. METHODS:The volatile oils of several Lonicera and Citrus TCM were determined by infrared spectroscopy. All samples of infrared spectrum were classified by hierarchical clustering,K-mean clustering,artificial neural networks,and support vector machine. RESULTS:The results of hierarchical clustering and K-mean clus-tering were ineffective. Methods of artificial neural networks and support vector machine achieved correct classification rate of 100%. CONCLUSIONS:Artificial neural networks and support vector machine can be combined with infrared spectroscopy to cre-ate chemometric fingerprinting for the identification of volatile oils of TCM.

Objective To set up a method for identification of Lonicerae japonicae flos volatile oils using Fourier-transform infrared spectroscopy. Methods The volatile oils of Lonicerae japonicae flos and Lonicerae flos was extracted by steam distillation combined with continuous liquid-liquid extraction with hexane. An oil film was prepared for Fourier-transform infrared spectroscopy scanning by dropping the volatile oils solution on the KBr disc and evaporating the solvent. The obtained infrared spectrum was treated by baseline removing and median filter smoothing. The spectral data within 1800-850 cm-1 was selected as the characteristic spectrum for hierarchical cluster analysis. And the volatile oils of Lonicerae japonicae flos and Lonicerae flos were discriminated by the result of hierarchical cluster analysis. Results Enough volatile oils were extracted for obtaining Fourier-transform infrared spectrum from small amount of Lonicerae japonicae flos. The method developed in the study was able to discriminate Lonicerae japonicae flos volatile oils from Lonicerae flos volatile oils. Conclusion The method can be used for identification of Lonicerae japonicae flos volatile oils.

OBJECTIVE: To investigate the pathogens and their resistance in continuous ambulatory peritoneal dialysis (CAPD) related peritonitis. METHODS: A total of 78 cases with CAPD related peritonitis from Xiangya Hospital between January 2007 and January 2011 were reviewed. Pathogens, resistance and outcomes of the 78 cases CAPD related peritonitis were analyzed retrospectively. RESULTS: Among them, 53 cases cultured positive (66.67%), 3 of which were combined infection and 2 strains were cultured. A total of 55 strains were cultured, including 32 gram-positive strains (58.18%), 18 gram-negative strains (32.72%) and 5 fungi (9.09%). The most common pathogens were coagulase negative staphylococcus, staphylococcus aureus and Escherichia coli. Drug sensitivity test of the gram-positive strains showed that the three with lowest antibiotic resistance were linezolid (0), teicoplanin (3.13%) and vancomycin (4.0%). Drug sensitivity test of the gram-negative bacteria showed that antibiotics with the lowest resistance were amikacin and imipenem, followed by piperacillin/tazobactam and cepoperazon/sulbactam. Cephazolin had the highest resistance rate of 83.33%. Clinical outcomes: 63 cases cured (80.77%); 11 cases transferred to hemodialysis (14.1%); 4 cases died (5.13%), including 2 cases fungus infections, 1 gram-negative bacteria infection and 1 combined infection. CONCLUSION The most common pathogens causing peritoneal dialysis associated peritonitis is gram positive bacteria. In the empirical treatment, in addition to traditional treatment of Cefazolin combined with aminoglycosides, cefazolin combined with piperacillin/tazobactam or cepoperazon/sulbactam is preferable for CAPD associated peritonitis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; drug effects ; isolation & purification ; Child ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Peritoneal Dialysis, Continuous Ambulatory ; adverse effects ; Peritonitis ; etiology ; microbiology ; Young Adult

OBJECTIVE To investigate the effect of injection of β2-adrenergic receptor agonist clenbuterol into the infralimbic cortex(IL) on drug-seeking behavior triggered by conditioned cues. METHODS Adult male SD rats were trained to self-administer heroin under a FR1 schedule for consecutive 14 d,followed by 2-h extinction training. Cue-induced heroin seeking was measured for 2 h. Clenbuterol was microinjected bilaterally into the IL(8 ng/side)of rats 15 min prior to reinstatement test. Meanwhile,locomotor activity was detected 15 min after clenbuterol or artifial cerebrospinal fluid(mod?el group) was microinjected bilaterally into IL. Western blotting was used to detect the expression of phosphorylated cyclic AMP response element-binding protein(p-CREB)in the prelimbic cortex(PL), IL,nucleus accumbens core (NACc) and shell (NACsh) of rats immediately after reinstatement test. RESULTS After heroin administration training for 14 consecutive days,these animals exhibited reliable heroin self-administration,indicated by the increase in active nose poke responses and infusions. The rats that had received infusion of clenbuterol into the IL had significantly lower active pokes (8 ± 3)than those in model group(45±10)in cue-induced reinstatement(P<0.01),but there was no significant differ?ence between clenbuterol group and vehicle group in the locomotor activity. The expression of p-CREB in either IL or NACsh was significantly decreased in clenbuterol group compared with model group(P<0.01,P<0.05),but significantly increased in NACc(P<0.01). CONCLUSION Microinjection of clenb?uterol into the IL can attenuate the cue-induced reinstatement of heroin-seeking behavior in rats. The underlying mechanism might be related to the regulation of p-CREB expression in the NACc and NACsh.

OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) extract and losartan (Los) on the expression of Klotho (Kl), P53, P21, and apoptosis in renal tubular epithelial cell NRK-52E induced by angiotensin II (Ang II), and to elucidate its therapeutical mechanism in Ang II induced renal tubular epithelial cell apoptosis. METHODS: NRK-52E cells were incubated with CS with or without Ang II for 24 hours. Experimental groups were divided according to the increasing concentrations of CS:0 (serving as controls), 5, 10, 20, 40, and 80 mg/L. The optimal concentration of CS was selected and cells were divided into 5 groups: controls, Ang II (1*10(-8) mol/L), Ang II (1*10(-8) mol/L)+CS (40 mg/L), Ang II (1*10(-8) mol/L)+Los (1*10(-5) mol/L), and Ang II (1*10(-8) mol/L)+CS (40 mg/L)+Los (1*10(-5) mol/L). After 24 hours, cell proliferation was evaluated by MTT assay. The mRNA and protein expression of Kl, P53 and P21 were measured by RT-PCR. Activity of caspase-3 was evaluated by caspase-3 activity assay Kit. Cell apoptosis was determined by Annexin V-FITC/PI double staining and flow cytometry. RESULTS: Certain concentrations of CS promoted the proliferation of NRK-52E cells and increased cells proliferation inhibited by Ang II (P<0.01 or P<0.05 ). Ang II significantly down-regulated the mRNA and protein expression of Kl, and up-regulated the levels of P53 and P21. Caspase-3 activity and apoptotic rates were decreased, too (all P values<0.01). CS or/and Los significantly increased the expression of Kl mRNA and protein down-regulated by Ang II, decreased P53 mRNA and protein expression, P21 mRNA and protein expression,and inhibited caspase-3 activity and apoptotic rates(all P values<0.05). No cooperative effects were observed in the two drugs (P>0.05). CONCLUSION CS can increase the expression of Kl down-regulated by Ang II, decrease P53 and P21 expression and caspase-3 activity, and reduce Ang II induced NRK-52E cell apoptosis, which may be part of its mechanism of the protective effects on hypertensive renal damage.
Angiotensin II ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Cordyceps ; chemistry ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; metabolism ; Glucuronidase ; genetics ; metabolism ; Humans ; Kidney Tubules ; cytology ; metabolism ; Losartan ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

ObjectiveTo study the effect of total flavone of Litchi Semen （TFL） on proliferation， apoptosis， migration， and invasion of hepatoma cells HepG2. MethodMethyl thiazolyl tetrazolium colorimetric （MTT） assay was used to detect the effect of different-dose TFL and cisplatin on the proliferation of HepG2 cells. TdT-mediated dUTP nick-end labeling （TUNEL） assay was used to detect the effects of low， medium， and high-dose （70， 140， 210 mg·L^-1） of TFL and cisplatin （60 mg·L^-1） on the apoptosis of HepG2 cells， thus selecting the optimal dose of TFL for the follow-up experiment. HepG2 cells were divided into a blank group， a TFL group （140 mg·L^-1）， a TFL+XL019 group （140 mg·L^-1 TFL+0.5 μmol·L^-1 XL019）， and a TFL+TPI-1 group （140 mg·L^-1 TFL+1 μmol·L^-1 TPI-1）. The effect of TFL on migration and invasion of HepG2 cells were examined by wound healing test and Transwell invasion assay， and the effect of TFL on the expression of epithelial-mesenchymal transition （EMT） marker in HepG2 cells were examined by cell immunofluorescence assay. Western blot was used to detect the expression of key proteins in Janus kinase 2（JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway after the intervention by TFL. ResultMTT assay showed that the proliferation of HepG2 cells was significantly inhibited by TFL and cisplatin at 24 and 48 h as compared with blank group （P<0.01）， and the half maximal inhibitory concentration （IC₅₀） of TFL on HepG2 cells was （136.7±2.40） mg·L^-1 at 24 h and （106.8±1.11） mg·L^-1 at 48 h. The IC₅₀ of cisplatin on HepG2 cells was （58.48±2.04） mg·L^-1 at 24 h and （5.15±0.56） mg·L^-1 at 48 h. The results of TUNEL assay showed that TFL induced apoptosis of HepG2 cells. The optimal dose of TFL was 140 mg·L^-1. The results of wound healing test showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the migration of HepG2 cells （P<0.05， P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 Group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The Transwell invasion assay showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the invasion of HepG2 cells （P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The results of immunofluorescence showed the intervention of TFL up-regulated the expression of E-cadherin， and down-regulated the expression of Vimentin in HepG2 cells， which was stronger in the TFL+XL019 group and weaker in the TFL+TPI-1 group. The results of Western blot showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group did not affect the expression of JAK2 or STAT3 protein， but significantly decreased the expression levels of phosphorylatied （p）-JAK2 and p-STAT3 （P<0.05， P<0.01）. As compared with the TFL group， the expression levels of p-JAK2 and p-STAT3 in the TFL+XL019 group were significantly decreased （P<0.01）， while those in the TFL+TPI-1 group were significantly increased （P<0.01）. Compared with the blank group， the TFL group significantly increased the expression level of Src-homology domain 2 containing protein tyrosine phosphatase-1（SHP-1） with sh2 domain （P<0.01）. ConclusionTFL has the effects of inhibiting the proliferation， promoting apoptosis of HepG2 cells， and reversing the EMT process of HepG2 cells to reduce the migration and invasion， which are presumably related to the activation of SHP-1 by TFL to block JAK/STAT3 signaling pathway.