This study was designed to determine if the decrease in the expression of interleukin 2(IL 2) in spleen lymphocytes after trauma was associated with the changes in the transcription factor NFAT binding activity. Closed impact injury with fracture of both hind limbs in mice was adopted as the trauma model. Animals were sacrificed at the 12h and 1,4,7,10,14 days after injury. Spleen lymphocytes were isolated from traumatized mice and stimulated with concanavalin A, The culture supernatants were harvested and assayed for IL 2 activity. Total RNA was extracted from spleen lymphocytes and assayed for IL 2 mRNA. Nuclear protein was extracted,and DNA binding activity of NFAT was measured using an electrophoretic mobility shift assay(EMSA). The results showed that the DNA binding activity of NFAT gradually decreased , with trough value of NFAT binding activity ,accounting for 41% of the control, observed at 4d after injury. It was closely associated with the decline of IL 2 activity and IL 2 mRNA. These data suggest that the decline in the induction of IL 2 expression is,at least, partially due to the impairment in the activation of NFAT in traumatized mice. Taken together, the results are significant to clarify the mechanism of the decline in the induction of IL 2 expression after trauma.

AIM: To observe the curative effect of PDT combined with intravitreal injection of ranibizumab treatment for age-related macular degeneration with choroidal neovascularization ( CNV) .
METHODS:In accordance with the inclusion criteria, by indocyanine green choroidalangiography ( ICGA ) and optical coherence tomography ( OCT ) examination confirmed the diagnosis of macular CNV in 27 patients (27 eyes), treated with PDT 3 ~ 7d professional intravitreal injection of ranibizumab. At 1, 3, 6mo after treatment, the results of best corrected visual acuity (BCVA), FFA, ICGA, OCT examination and complications were observed.
RESULTS: The BCVA improved in 17 eyes ( 63%) , stable in 6 eyes ( 22%) , and decreased in 4 eyes ( 15%) . Before treatment, the average leakage area was 1 005. 69±105. 47μm, it were 875. 54 ± 103. 27μm, and 423. 37 ±79.68μm at 1 and 3mo after treatment, there were significant differences compared with before treatment ( P<0. 01). Average central macular thickness of retina before treatment was 485. 58±122. 59μm, and 398. 84±105. 32μm, 297. 74±89. 18μ m at 1 and 3mo after treatment, there were significant differences compared with before treatment( P<0. 01).
CONCLUSION: The method that PDT closed CNV combined with intravitreal injection of ranibizumab can effectively block angiogenesis recurrence, reduce the number of PDT treatment again and complications, improve the therapeutic effect.

ObjectiveTo investigate the distribution of high-intensity zone (HIZ) of lumbar intervertebral disc in patients with low back and/or leg pain,and analyze its related factors.MethodsSix hundred and twenty-eight patients with low back and/or leg pain were examined by MRI scan from June 2009 to August 2010.According to the diagnostic criteria of HIZ,the features of distribution of HIZ on age,segment and degree of intervertebral disc degeneration were analyzed retrospectively.ResultsAmong 3140 intervertebral discs of the 628 patients,172 cases (27.39％,172/628) and 206 discs (6.56％,206/3140)were involved with HIZ.There was no significant difference between men and women [26.38％(86/326)vs.28.48％(86/302)] (P=0.556).HIZ occurred more often [40.22％(72/179)] in those patients between 40 and 49 years of age.The incidence of HIZ at the segments from L1-2,L2-3,L3-4,L4-5,L5-S1 was 0.80％(5/628),2.07％(13/628),2.07％(13/628),14.01％(88/628) and 13.85％(87/628) respectively.In cases with and without HIZ,the incidence of intervertebral disc degeneration up to grade V was 49.03％(101/206) and 23.76％(697/2934) respectively (P ＜ 0.01 ).HIZ was correlated with age,degree of intervertebral disc degeneration and disc segment (r =-0.040,P=0.025 ;r =0.217,P＜ 0.01 ;r =0.179,P＜ 0.01 ).Conclusions HIZ is correlated with age,degree of intervertebral disc degeneration and disc segment.Intervertebral disc degeneration plays the most important role in the occurence of HIZ.HIZ_mainly occur in L4-5 and L5-S1 segment and in those between 40 and 49 years of age.

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

Objective To investigate the prognostic factors regarding overall survival in locally advanced stage of esophageal squamous cell carcinoma (ESCC) patients after receiving concurrent chemoradiotherapy (CRT).Methods Each of 80 patients with locally advanced ESCC was treated by doxtel 135-175 mg/m2 d1 and cisplatin 40 mg/m2 d2,3 combined with radiotherapy 60 Gy.ECOG (0-1),tumor locations,tumor sizes,TNM classifications as well as the levels of CA199,SCC-Ag were detected before concurrent treatments and 4 weeks after treatment.The relationship wasanalyzed between the prognosis of disease and CA199,SCC-Ag levels.Each index was studied by Cox statistical regression analysis and P value was determined.Results All patients had completed 4 cycles' treatments successfully.The major adverse effects included neutrophilic granuloaytopenia,calvities,nausea,emesis and diarrhea.The severe effect was Ⅲ degree neutrophilic granuloaytopenia for 2 cases (2.5 ％),all the side-affect had recovered after symptomatic treatments.The median survival time were 50 months and 72.5 months in the patients who were greater than 60 year-old and less than or equal to 60 year-old,respectively (P =0.004).Regarding to 5-year overall survival,the figures were 73.3 ％,69.4 ％,41.7 ％ and 0 in the patients with cT1-2N1M0,cT3N1M0,cT4NoM0 and cT4N1M0 classification,respectively (P =0.024),they were 42.9 ％,70.0 ％,and 60.9 ％ in the patients with upper thoracic esophagus,middle thoracic esophagus,and lower thoracic esophagus,respectively (P =0.971),they were 100 ％ in the patients with ECOG 0 and 54.1％ in the patients with ECOG 1 (P =0.044),they were 86.7 ％ in the patients with CA199≤37 kU/L,and 52.0 ％ in the patients with CA199 more than 37 kU/L (P =0.008),they were 95.5 ％ in the patients with SCC-Ag≤ 1.9 μg/L,and 53.4 ％ with ＞1.9 μg/L (P =0.012).Conclusion For ESCC patients treated with CRT,the age,TNM stage,ECOG,CA199 and SCC-Ag levels are independent forecasting factors regarding their overall survival.

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

OBJECTIVETo explore the risk factors of ventilator-associated pneumonia (VAP) in patients admitted in an intensive care unit (ICU) for pulmonary tuberculosis (TB).

METHODSThe clinical data of 143 patients admitted in the ICU at our center between January, 2014 and June, 2015 were reviewed. The patients with VAP and those without VAP were analyzed for risk factors of VAP in the setting of an ICU for pulmonary TB and compared for the duration of ventilation and hospital stay.

RESULTSThe patients with pulmonary TB showed a significantly higher incidence of VAP in the ICU than those without TB. Univariate analysis suggested that the occurrence of VAP was significantly correlated with the duration of mechanical ventilation, invasive examination, pulmonary tuberculosis, lung structure changes, use of multiple antibiotics, diabetes, tracheal incision, indwelling gastric tube, APACHE II score, and coma (P<0.05). Multivariate logistic regression analysis showed that pulmonary TB, duration of mechanical ventilation, APACHE II score, invasive operation, and use of multiple antibiotics were independent risk factors for VAP (P<0.05). The patients who developed VAP had a prolonged duration of mechanical ventilation and ICU stay (P<0.05).

CONCLUSIONPatients admitted in tuberculosis ICU are exposed to a high risk of VAP with a high mortality rate as the result of multiple interacting risk factors. Pulmonary TB, prolonged mechanical ventilation, an APACHE II score >15, invasive operation, and use of multiple antibiotics are all independent risk factors for VAP in tuberculosis ICU.

APACHE ; Anti-Bacterial Agents ; adverse effects ; Humans ; Incidence ; Intensive Care Units ; Length of Stay ; Pneumonia, Ventilator-Associated ; complications ; Respiration, Artificial ; adverse effects ; Risk Factors ; Time Factors ; Tuberculosis, Pulmonary ; complications

OBJECTIVETo study the effect of flavonoids from seed residues of Hippophae rhamnoides (FH) on the lipid metabolism and antioxidative activity in climacteric rats.

METHODMenopausal rats with aging were used in this experiment. The rats were fed with FH by gastrogavage for 13 weeks. The effect of drug on the lipid metabolism and the antioxidative activity were observed after the rats were killed.

RESULTSerum total cholesterol was decreased significantly in rats fed with FH, T-AOC and SOD in serum and liver were significantly higher than those in rats fed with water, and at the same time MDA was lower than that in rats fed with water.

CONCLUSIONFH can improve the climacteric rats' lipid metabolism, and enhance the antioxidation in climacteric rats.

Animals ; Antioxidants ; pharmacology ; Cholesterol ; blood ; Climacteric ; blood ; Female ; Flavonoids ; isolation & purification ; pharmacology ; Hippophae ; chemistry ; Lipid Metabolism ; drug effects ; Liver ; metabolism ; Malondialdehyde ; blood ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry ; Superoxide Dismutase ; blood ; metabolism ; Triglycerides ; blood

Airway remodeling is an important pathological feature of asthma and the basis of severe asthma. Proliferation of airway smooth muscle cells (ASMCs) is a major contributor to airway remodeling. As an important Ca(2+) channel, transient receptor potential vanilloid 1 (TRPV1) plays the key role in the cell pathological and physiological processes. This study investigated the expression and activity of TRPV1 channel, and further clarified the effect of TRPV1 channel on the ASMCs proliferation and apoptosis in order to provide the scientific basis to treat asthmatic airway remodeling in clinical practice. Immunofluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of TRPV1 in rat ASMCs. Intracellular Ca(2+) was detected using the single cell confocal fluorescence microscopy measurement loaded with Fluo-4/AM. The cell cycles were observed by flow cytometry. MTT assay and Hoechst 33258 staining were used to detect the proliferation and apoptosis of ASMCs in rats respectively. The data showed that: (1) TRPV1 channel was present in rat ASMCs. (2) TRPV1 channel agonist, capsaicin, increased the Ca(2+) influx in a concentration-dependent manner (EC50=284.3±58 nmol/L). TRPV1 channel antagonist, capsazepine, inhibited Ca(2+) influx in rat ASMCs. (3) Capsaicin significantly increased the percentage of S+G2M ASMCs and the absorbance of MTT assay. Capsazepine had the opposite effect. (4) Capsaicin significantly inhibited the apoptosis, whereas capsazepine had the opposite effect. These results suggest that TRPV1 is present and mediates Ca(2+) influx in rat ASMCs. TRPV1 activity stimulates proliferation of ASMCs in rats.

BACKGROUNDRetinal light injury can lead to degeneration of the photoreceptor cell layer. It has been hypothesized that the mechanism for this process is the photochemical damage. Ginkgo balboa extract (Ginkgo biloba extract EGB761) EGB761 is a free radical scavenger. The purpose of this study was to investigate the possible effect of orally administered EGB761 on retinal light damage of mouse photoreceptor cells.

METHODSKunming mice were randomly chosen for the following groups containing 20 animals in each: control group, light damage group, saline control group, and drug treatment group. The drug treatment group and saline control group were given daily gavage of EGB761 (150 mg×kg(-1)×d(-1)) one week before light exposure. At 7, 14, and 30 days after light exposure, animals were sacrificed and eyes were examined by light microscopy, electron microscopy, and retinal histopathology using in situ detection of apoptotic cells.

RESULTSIn the light damage group after 7 days there was visible edema, and the outer nuclear layer appeared withered with deeply stained dead cells, leaving only a thin nuclear layer of 7 - 8 cells. After 14 days, the photoreceptor cell layer disappeared, leaving only the outer nuclear layer of 1 - 3 cells with an average thickness of (37.988 ± 1.207) µm. The average thickness of the retina was (126.32 ± 2.31) µm. In the drug treatment group, the photoreceptor cell layer and outer nuclear layer damage were significantly lower than the saline group (t = 21.993, P < 0.001), demonstrating that EGB761, especially at 14 days after light exposure, can reduce retinal light damage in mice.

CONCLUSIONOral administration of EGB761 can partially inhibit apoptosis of photoreceptor cells, resulting in increased photoreceptor cell survival.

Animals ; Eye Injuries ; etiology ; prevention & control ; Light ; adverse effects ; Male ; Mice ; Microscopy, Electron ; Photoreceptor Cells ; drug effects ; radiation effects ; ultrastructure ; Plant Extracts ; therapeutic use ; Rats ; Retina ; drug effects ; radiation effects ; ultrastructure