Objective To investigate the effect of surfactant protein A (SP-A) on the production of MIP-2 and binding activity of NF-?B in rat tubular epithelial cells, and evaluate its possible role in renal inflammation. Methods Confluent cultures of NRK-52E cells (a renal tubular epithelial cell line of rat origin) were pretreated with various concentrations of SP-A(0 to 80 ?g/ml) and stimulated by lipopolysaccharide (LPS) (10 ?g/ml) with 2% serum. MIP-2 expression was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The effect of SP-A on NF-?B binding activity was assessed by electrophoretic mobility shift assay (EMSA). Results MIP-2 mRNA and protein was expressed and up-regulated in NRK-52E cells stimulated by LPS. The expression of MIP-2 was down-regulated by SP-A. NF-?B binding activity was inhibited by SP-A in a concentration-dependent manner. Conclusion SP-A binding activity and down-regulates the expression of MIP-2 in renal tubular epithelial cells, which may play an important role in the modulation of renal tissue inflammation.

Objective To evaluate the effect of ALD on podocyte apoptosis and the possible roles of Akt in ALD-induced apoptosis. Methods The cultured rat podocytes were incubated with increasing concentrations of ALD (10-9~10-5 mol/L) for variable time periods. Apoptosis was evaluated by cell nucleus staining and flow cytometry. RT-PCR was used to examine the expression of mineralocorticoid receptor (MR)and 11 Beta-hydroxysteroid dehydrogenase type 2 (11?-HSD2) mRNA in podocyte. Activation of Akt/PKB was evaluated by performing Akt kinase assay. Results ALD induced podocyte apoptosis in a dose- and time-dependent manner. The proapoptotic effect was attenuated by the presence of spironolactone (10-7mol/L). The expression of MR and 11P-HSD2 mRNA was demonstrated in the podocytes by RT-PCR. ALD also inhibited the activity of Akt in a dose-dependent manner, but the inhibitory effect was significantly ameliorated by the presence of spironolactone. The activity of Akt was negatively correlated with podocyte apoptosis. Conclusion ALD induces apoptosis in rat podocytes through the signaling mechanism by which Akt is inhibited.

Objective Fibroblast-like synoviocytes (FLS) play important roles in the pathogenesis of rheumatoid arthritis (RA).The present study was undertaken to investigate the mechanism of calreticulin (CRT) to promote FLS survival in RA.Methods FLS were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and osteoarthritis (OA) patients and cultured in vitro.The expression of Bcl-XL and Mcl-1 in FLS at mRNA and protein level was detected by quantitative-polymerase chain reaction (q-PCR),Western blotting and immunofluorescence respectively.RA and OA FLS were cultured with different concentrations of recombinant human CRT for 48-72 h,the expression of Bcl-XL and Mcl-1 was detected by q-PCR and Western blotting.The proliferation of RA FLS following CRT stimulation was determined by MTT assay.Results ① Compared with FLS from OA patients (1.00±0.39;1.00±0.46),the anti-apoptotic Bcl-XL and Mcl-l mRNA expression (14.51 ±2.20;12.82±1.80) was significantly higher in the FLS from RA patients (t=10.47,1 1.02;P＜0.01);Western blotting analysis also showed increased protein levels of Bcl-XL and Mcl-1 in RA FLS;Immunofluorescence results showed higher expression of Bcl-XL and Mcl-1 in RA at the single FLS level;② CRT up-regulated the expression of Bcl-XL and Mcl-1 in RA FLS:compared with the control group (0 ng/ml),CRT stimulation at 10 ng/ml and 50 ng/ml increased the levels of Bcl-XL mRNA (1.70±0.28 vs 1.00±0.20,q=4.58,P＜0.05;1.87±0.35 vs 1.00±0.20,q=5.69,P＜O.05) and Mcl-1 mRNA (1.85±0.36 vs 1.00±0.20,q=5.63,P＜0.05;1.72±0.26 vs 1.00±0.20,q=4.77,P＜0.05) in RA FLS,while no significant effects of CRT on Bcl-XL and Mcl-1 mRNA expression were observed in OA FLS (F=1.49,1.60;P＞0.05);Western blotting results showed elevated protein levels of both Bcl-XL and Mcl-1 in RA FLS after CRT treatment at a concentration dependent manner.However,neither Bcl-XL nor Mcl-1 expression was significantly changed in OA FLS.③ MTT assay showed that CRT had no significant effect on the proliferation of RA FLS (F=2.88,P＞ 0.05).Conclusion Our results indicate that CRT-mediated up-regulation of anti-apoptotic Bcl-XL and Mcl-1 may inhibit apoptosis and promote the survival of FLS from RA patients.

OBJECTIVETo introduce the framework of evidence-based practice with a case of castration-resistant prostate cancer (CRPC) as an example.

METHODSA clinical question was formulated according the clinical scenario. A systematic search was conducted for the published literature in the databases of PubMed, EMBASE, Cochrane Library, Clinical Trial Registries, and Web of Knowledge up to Dec 2014. The identified literature was reviewed for quality appraisal before the evidence was applied to clinical practice.

RESULTSThe treatment was effective and the patient achieved disease remission.

CONCLUSIONEvidence-based practice should be integrated with clinical scenario, current evidence, and patients' willingness, and follow a systematic framework.

Evidence-Based Medicine ; Humans ; Male ; Orchiectomy ; Prostatic Neoplasms, Castration-Resistant ; therapy

Objective To understand the heterochro-Matin-associated protein 1(HP1-γ) expression during the carcinogenesis and progresston of esophageal carcmoma,and preliminarily investigate the supertortty of using laser scanning cytometer to analyze immunohistochemistry results compared to traditional scoring methods.Methods A tissue microarray containing different grades of esophageal carcinoma was selected and the HP1-γ expression was detected by immunohistochemistry.The results of immunohistochemistry were analyzed quantitatively by using laser scanning cytometer.The correlation of results analyzed by using laser scanning cytometer and traditional scoring methods was analyzed by chi square test.Results The HP1-γ was primarily expressed in the nucleus.The positive rate of HP1-γ in normal esophagus,moderate-severe atypical hyperplasia，in situ carcinoma and squamous cancer was 37.5% (3/8),100%(21/21),100%(7/7) and 23.7% (9/38),respectively,with the difference being statistically significant among normal esophagus，oderate-severe atypical hyperplasia plus in situ carcinoma and squamous cancer(P<0.01).There was a high correlation between the results analyzed by laser scanning cytometer and those by traditional scoring methods under a light microscope(P<0.01).Conclusion HP1-γ may play a resistant role in the transformation from normal esophageal cells to malignant cells.Compared to the traditionally artificial scoring methods,there are many advantages such as high resolution,high objectivity and accuracy when using laser scanning cytometer to analyze the immunohisto chemistry results.

Adult ; Carcinoma, Renal Cell ; secondary ; Female ; Humans ; Kidney Neoplasms ; pathology ; Magnetic Resonance Imaging ; Vaginal Neoplasms ; secondary

OBJECTIVETo discuss the hemostasis of the different polarities of Platycladi Cacumen Carbonisatum (PCC) on the blood heat and hemorrhage syndrome rat model induced by dry yeast.

METHODThe SD rats were divided into seven groups. Yunnan Baiyao was taken as the positive control drug. The rats in the control group and model group were fed with CMC-Na for 7 days, and the rats in other groups were fed with corresponding drugs simultaneously. On day 7, the blood heat and hemorrhage syndrome rat model was established. Indexes including the whole blood viscosity, plasma viscosity, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen content (FIB), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), blood platelet count (PLT), thrombocytocrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) and the rate of platelet aggregation induced by ADP were detected. Additionally, the pathological examinations of lungs among each group were compared.

RESULTCompared with the control group, the RBC, HGB and HCT of rats in the model group increased significantly, with distinct increase in high, middle and low whole blood viscosity and plasma viscosity of rats in the model group; TT and APTT were notably prolonged, while PT was notably shortened, with significant increase in FIB content; PLT, PCT, MPV and PDW remarkably increased; Additionally, the rate of platelet aggregation induced by ADP significantly decreased. After ig administration of the ethyl acetate extract of PCC, the low whole blood viscosity and plasma viscosity remarkably decreased; TT and APTT were significantly shortened, with notable reduction in PDW and in FIB content Additionally, the rate of platelet aggregation induced by ADP significantly increased. The injury of lungs was also improved in ethyl acetate extract group. The rate of platelet aggregation induced by ADP of n-butanol extract group notablly increased. Plasma viscosity of water extract group remarkably decreased, with TT being significantly shortened. But the effects of n-butanol extract or water extract were weaker than that of ethyl acetate extract. And the effect of petroleum ether extract was the weakest.

CONCLUSIONEthyl acetate extract is the active part of PCC, showing the effect of hemostasis by reducing the low whole blood and plasma viscosity, improving coagulation function mainly by acting on the endogenous coagulation, and ameliorating the function of platelet aggregation.

Animals ; Blood Coagulation ; drug effects ; Blood Viscosity ; drug effects ; Cupressaceae ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Filtration ; Hemostatics ; isolation & purification ; pharmacology ; Male ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Thrombin Time

This article was aimed to study effect of D-ribose on the high-energy phosphate metabolism of skeletal muscle tissues of tired mice. The model was made by burden swimming. And then, the mice were divided into four groups, which were the model group, D-ribose group, caffeine group, and D-ribose with caffeine group). Intragastric administrations of drugs were given to all mice in four groups, three times per day. And all mice continued to swim for three days. The time of swimming was recorded. Gastrocnemius of mice were removed after swimming or 3 days later to measure the concentration of ATP, ADP, AMP and IMP with the HPLC. The results showed that compared with the control group, the time of burden s wimming was significantly prolonged for mice in the D-ribose group and the D-ribose with caffeine group. After three-day recovery, the concentration of ATP, AMP and IMP of gastrocnemius in the D-ribose group and the D-ribose with caffeine group mice was significantly increased. There was no significant difference in the caffeine group mice. It was concluded that D-ribose is involved in the high-energy phosphate metabolism of skeletal muscle tissues of tired mice . D-ribose promotes the recovery of ATP concentration in the gastrocnemius of tired mice, and prolongs the time of burden swimming. Therefore, it has a certain anti-fatigue effect .

BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy.
OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro.
METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal
interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours.
RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (P<0.05), and the inhibition effect was more significant at 72
hours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ,
feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after
trasfection (P<0.05), and the inhibition effect was more significant at 72 hours after transfection when compared with that at 48 hours after transfection, which was same with the effect on mRNA level. RNA interference in vitro can
significantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.

Objective To evaluate the changes of urodynamics in female patients with diabetic cystopathy.Methods Fifty six female patients with diabetic cystopathy were enrolled in the study , including 31 cases with diabetic course <15 years ( groupⅠ) and 25 cases with diabetic course ≥15 years ( group Ⅱ) .Urodynamic examination was performed in all patients and the urodynamic parameters were compared between two groups.Results The average residual urine volume , the volume of first bladder sensation and the max bladder capacity in groups ⅠandⅡwere (35 ±16)ml and (65 ±24) ml,(220 ± 76)ml and (330 ±88) ml, (380 ±92) ml and (580 ±122) ml, respectively; 3 out 31 and 16 out of 25 patients in groups Ⅰ and Ⅱ showed low compliance .The above indexes between two groups were statistically significant ( P<0.01).Conclusion Urodynamics can indicate the severity of functional damage of urinary bladder in patients with diabetic cystopathy .