?AlM: To explore the effects of the prescription of reinforcing kidney, nourishing blood, improving eyesight on the oxidative stress model of ARPE-19 cells induced by acrolein.
?METHODS:SD rats serum containing the prescription of reinforcing kidney, nourishing blood, improving eyesight and the content of distilled water in serum were prepared. The effects of the prescription and distilled water in serum at different concentration ( 2. 5%, 5%, 10%, 20% and 40%) on cell vitality was observed by cell counting kit ( CCK-8 ) assay. the logarithmic phase of ARPE-19 cells were pretreated by different concentrations (1. 25%, 2. 5%and 5%) of the prescription serum and distilled water in serum for 24h. Then it was treated with 75μmol/L acrolein for 24h. Cell vitality was observed by CCK-8 assay. The change of cell nucleus was detected by DAPl staining .
?RESULTS: 2. 5% and 5% serum had no effect on cell viability (P>0. 05), while 10%, 20%, 40% serum could inhibit cell viability (P<0. 05). CCK-8 results showed that 2. 5% and 5% the prescription of reinforcing kidney, nourishing blood, improving eyesight serum was better than distilled water in serum (P<0. 05).
?CONCLUSlON: The prescription of reinforcing kidney, nourishing blood, improving eyesight has the protective effect on ARPE-19 cell damage induced by acrolein.

Objective Present study aimed to observe the effects of Apoptin gene on killing retinoblastoma HXO-RB_(44) cells and illustrates its mechanisms. Methods Human retinoblastoma cells strain, HXO-RB_(44), was cultured and passaged in RPMI 1640 medium containing bovine serum. Apoptin gene was transfected into HXO-RB_(44) cells by liposome into HXO-RB_(44)/Apoptin, and pcDNA_3 was transfected in HXO-RB_(44)/peDNA_3 group. The expression of Apoptin mRNA was detected using Reverse Transcription Polymerase Chain Reaction (RT-PCR). The expression of protein of Apoptin and p53 were detected by SABC immunohistochemistry. The growth rate of HXO-RB_(44) cells was studied by constructing the growth curve and calculated as the formula: inhibitory rate = 1-cell number in experiment group/cell number in control group x 100%. Cellular apoptosis was determined by flow cytometry. Results The RT-PCR result showed the 450 kb specific band in UXO-RB_(44)/Apoptin group and absent amplification result in HXO-RB_(44) group and HXO-RB_(44)/pcDNA_3 group. The difference in SABC-positive cell number between HXO-RB_(44)/Apoptin group and control group was statistically significant (P ＜ 0. 05). The growth of HXO-RB_(44) cells was significantly inhibited in HXO-RB_(44)/Apoptin group compared with control group (P ＜ 0.05). Apoptosis cells increased significantly. The apoptosis rate was 38. 5% . Conclusion Apoptin gene could inhibit the growth of HXO-RB_(44) cells effectively. Up-regulation of expression of p53 gene might not be one of cell apoptosis mechanisms.

The aim of the present study was to assess whether Fourier transform infrared spectrometry (FTIR) micro-spectroscopy could produce distinct spectral information on protein of old myocardial infarction (OMI) and to set them as molecular markers to diagnose atypical OMI. Paraffin-embedded heart samples were derived from victims dying of OMI. In combination with histological stain, FTIR and infrared micro-spectroscopy, the characteristics of OMI were analyzed morphologically and molecularly. The most relevant bands identified were the amide A, B, I and, II showing crucial spectral differences between apparent normal region and OMI region, including the peak position blue shift and the increased intensity of OMI, moreover relative increase in alpha-helix and decrease in beta-sheet of protein secondary structures in OMI. Comparing to single spectral band, the I1650/I1550 ratio was increased and rationally used as a molecular marker for diagnosing OMI. These novel preliminary findings supported further exploration of FTIR molecular profiling in clinical or forensic study, and were in accordance with histopathology.

Objective:To study the effects of SiO2 nanoparticles on the organs of mice in vivo after intratracheal instillation, and to provide the basis for safety evaluation of SiO2 nanoparticles. Methods:Forty female BALB/c mice aged 6-8 weeks were randomly divided into control group (saline), low dose of SiO2 group (7 mg·kg-1), middle dose of SiO2 group (21 mg·kg-1), and high dose of SiO2 group (35 mg·kg-1).1 and 15 d after five times of non-exposed intratracheal instilation infection (once every 3 d), the mice were sacrificed and the left lungs,the right kidneys, livers, hearts and spleens were collected and embedded in paraffin.The morphology of tissue sections was observed under light microscope after hematoxylin-eosin (HE) staining.The eyeball blood was obtained and the biochemical indicators of liver and kidndy functions were detected.Results:Compared with control group, there were alveolar interval thickening, inflammatory cell infiltration, and a small amount of small arterial thrombosis in the lungs;granulomatous inflammatory cell infiltration and a small amount of focal necrosis of liver cells in the livers;red pulp enlargement, hyperemia, and more visibly scattered megakaryocytes in the spleens in SiO2 nanoparticles groups in a dose-dependent manner, especially in middle and high doses of SiO2 groups.After 15 d of injection, the damages alleviated with the prolongation of time.There was some inflammatory cell infiltration in the kidney tissue of the mice in SiO2 nanoparticle groups.The biochemical indicator detection results showed that alanine aminotransferase(ALT) and aspartate transaminase(AST) levels in SiO2 nanoparticles groups varied, suggesting the liver cell damages were at different degrees;the changes of urea nitrogen(BUN) and creatinine(Cr) levels in SiO2 nanoparticle groups remindered the kidney function alteration, but there were no obvious dose-and time-dependent effects.Conclusion:Intratracheal instillation of SiO2 nanoparticles can influence the major organs of the mice and mainly displays in the inflammation and injuries in the lung, liver, and spleen.

Objective To investigate whether there is amyloid fibers in the mature neutrophils and to reveal the molecular mechanism of the formation of AD amyloidosis. Method Thirty cases of AD patients and 30 healthy control were enrolled. The white blood cell, the red blood cell,hemoglobin and neutrophil absolute value were determined by Sysmex X5-500i hematology analyzer. The neutrophils and amyloid variable specific probe affinity were measured, The starch like variable fluorescence intensity in plasma was dtected by the sulfur T (Thioflavin T, ThT) method. Results The affinity test results showed that the amyloidosis fluorescent probe (ThT) can be combined with tissue amyloid fibers specifically. The neutrophil amyloid fibers staining also showed a positive reaction. Compared with the AD group, no significant differences were found in white blood cell, red blood cell, hemoglobin and neutrophil absolute value level in the healthy control group The serum amyloid variable fluorescence intensity in the AD group was significantly higher than that in the control group (P < 0.05). Conclusion The amyloid fibers was found in the mature neutrophils, and the level of plasma amyloid fibers was significantly increased in the AD patients.

The objective of this study is to develop a sensitive and reliable high-performance liquid chromatography mass spectrometry (LC-MS) method for simultaneous determination of artemisinin, arteannuin B, artemisic acid, and scopoletin, and study the pharmacokinetics of the four constituents in mouse serum after oral administration of the four components to mice. The analytical column used was Agilent Zorbax SB-C18 (2.1 mm x 150 mm, 5 mm). The mobile phase was acetonitrile: 0.5% acetic acid (60: 40) and the flow rate was 0.3 mL x min(-1). The temperature of the column was 40.0 degrees C. In this condition, we established an analysis method to simultaneously determine the four components. A sensitive and specific liquid chromatography-mass spectrometric (LC-MS) method was developed and validated for the determination of artemisin in derivatives in mice plasma. The method we established has a linear range of 5-3 000 μg x L(-1) with a good sensitivity and specificity for all of the four components. This method is simple, rapid, accurate and suitable for the determination of the content of the four compounds.
Animals ; Artemisinins ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Dose-Response Relationship, Drug ; Male ; Mice ; Reproducibility of Results ; Scopoletin ; blood ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; methods

The physiological mechanism underlying the acupoint sensitization was evaluated systemically by using the method of electric physiology at spinal cord, medulla, and thalamus levels; the dynamic change of acupoint from the relative "silence" to the relative "activation" function was explained through the study on the dynamic process of acupoint sensitization; the biological process of the therapeutic effect of acupoint stimulation was illuminated through the research of the central mechanism underlining the dose effect relationship between the sensitive acupoint and the related brain area, thus scientific evidence for the functional link between the acupoint and internal organs as well as the nature of the acupoint were provided.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Moxibustion ; Nociceptors ; physiology ; Sensation ; Viscera ; innervation ; physiology ; Visceral Afferents ; physiology

OBJECTIVETo evaluate the clinical effects of single posterior debridement, bone grafting, internal fixation and local chemotherapy in treating thoracolumbar spinal tuberculosis.

METHODSFrom February 2009 to September 2012,11 patients with thoracolumbar spinal tuberculosis were treated by single posterior debridement, bone grafting, internal fixation and local chemotherapy. There were 7 males and 4 females, aged from 27 to 65 years old with an average of 53.7 years. The courses of disease was from 3 months to 2 years with the mean of 9 months. According to ASIA standard of spinal cord injury, 3 cases were grade C and 8 cases D. After treatment, clinical effects were evaluated by ASIA grade, visual analogue score (VAS) and Oswestry Disability Index (ODI); kyphosis Cobb angle change was observed by X-rays.

RESULTSEleven patients were followed up from 12 to 29 months with an average of 18 months. ASIA grade of spinal cord injury, 3 patients with grade C improved to grade D in 2 cases and grade E in 1 case 8 patients with grade D improved to grade E in 7 cases and unchanged in 1 case. VAS decreased from preoperative 6.10 ± 1.30 to 1.70 ± 0.80 at 3 d after operation (P < 0.05). ODI improved from preoperative (68.36 ± 10.41)% to (14.55 ± 8.99)% (P < 0.05) at 3 d after operation. Kyphotic Cobb angle was corrected from preoperative (22.64 ± 4.84)° to (4.27 ± 1.49)° (P < 0.05) on the 3rd day after operation, and angle loss was mild at final follow-up, there was no significant difference between postoperative at 3 d and final follow-up.

CONCLUSIONSingle posterior debridement, bone grafting, internal fixation and local chemotherapy for the treatment of thoracolumbar spinal tuberculosis can effectively remove the lesion, improve nerve function and correct deformity, has advantage of single incision, little trauma, and low recurrence rate. But it still need long-term and systemic treatment with anti-TB drugs.

Adult ; Aged ; Bone Transplantation ; Debridement ; Female ; Humans ; Internal Fixators ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; therapy

OBJECTIVETo investigate the effects of complement inhibiting component of Ephedra sinica on immunological inflammation following acute spinal cord injury (SCI) in rats.

METHODSThe complement inhibiting component of Ephedra sinica was isolated by multiple precipitation steps and thin layer chromatography, and then the activity was analyzed. Fifty healthy SD rats were selected and randomly divided into the control group and the experimental group, 25 in each group. Induction of SCI was performed following a modified Allen's weight-drop method. The complement inhibiting component from Ephedra sinica (15 mg/kg) dissolving in 5 mL normal saline was immediately administered by gastrogavage after SCI, once daily. Equal volume of normal saline was administered to rats in the control group by gastrogavage. Hematoxylin and eosin (H&E) staining and C3 immunohistochemical staining were performed in SCI tissue at 12 h, day 1, 3, 7, and 14 after SCI. C3 positive expressions and myeloperoxidase (MPO) activity were assessed. Intercellular adhesion molecule-1 (ICAM-1) mRNA expression level was evaluated by Real-time PCR technique.

RESULTSC3 positive expression, MPO activity, and ICAM-1 mRNA level were significantly weaker in the Ephedra sinica group than in the control group at all time points (12 h, day 1, day 3, day 7, and day 14 after SCI) (P < 0.01, P < 0.05).

CONCLUSIONSThere existed complement system activation following acute SCI. The complement inhibiting component of Ephedra sinica significantly reduced immunological inflammation after SCI, and played an important role in secondary SCI.

Animals ; Complement Activation ; drug effects ; immunology ; Complement Inactivating Agents ; pharmacology ; Ephedra sinica ; chemistry ; Inflammation ; immunology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; immunology ; metabolism ; pathology

This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism. Two hundred SD rats were randomly divided into four groups: normal group, sham-operated group, model group and EA+TMS group, and each group was then divided into five sub-groups in terms of the different time to start treatment post operation: 6, 12, 24, 48 and 72 h. Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R). After treatment for 14 d, the Morris water maze test was applied to examine the spatial learning and memory abilities of rats. In infarcted area, the expression of caspase-3 was immunohistochemically detected, and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA. The results showed that in EA+TMS group compared with model group at the same treatment time windows, the escape latency was substantially shortened, the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05). In the EA+TMS sub-groups, the escape latency was shortest, the expression level of caspase-3 lowest, and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h. It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3. The best time window is 24 h after perfusion treatment to ischemia.