AIM:To determine the optimal extraction of effective component from Swertia chirayita(Roxb.ex Flemi) Karsten in Tibet. METHODS: HPLC method was used to determine the yield rate and mango glucoside content,the preference extract methods,extraction solvents and extraction method of Swertia chirayita(Roxb.ex Flemi) Karsten were chose by parallel comparative method. RESULTS: 75% ethanol was used to extract medicine materials by Percolate,the flow rate was 4 mL/min/kg,collecting 6 times liquor after percolate. CONCLUSION: The extraction method is simple and convenient,furthermore the extract stability is good too.

Objective To modify the techniques for establishment of an abdominal working heart transplantation model in rats and to sum up the key factors to success.Methods A total of 180 12-week old Brown Norway rats ( donor) and Lewis rats ( recipient) were used in this study:50 BN rats and 50 Lewis rats for pilot experiment, and 40 BN rats and 50 Lewis rats for the formal experiment.The rat model of working heart heterotopic transplantation was adopted and estab-lished by Wiedemann’ s mode.We transplanted the heart from BN rats to Lewis rats and analyzed the survival rate, causes of death and histological changes of the heart ( HE staining) in this experiment.Results After exercise and modification, the survival rate was increased to 77.5%, and the mean total duration of operation was 71 ±11 min, and the mean ische-mic time of the donor hearts was 34 ±5 min.Histological examination ( HE staining) of the cardiac allograft showed a mild inflammatory cell infiltration in the graft at 24 h after transplantation, indicating that the model was reliable.Conclusions A variety of factors may affect the final operation success rate in the establishment of this heart transplantation model.A-mong them, the major affecting factors include: healthy animals, donor heart protection, rapid and effective vascular su-ture, and postoperative animal management.

Objective To investigate the efficiency of target gene transfection of the heart and liver after tail vein or intramyocardial injection of adenovirus vector (GFP-Ad).Methods GFP-AD was constructed at first.A total of 20 male 8-week old C57BL/6 mice were randomly and equally divided into tail vein injection of GFP-AD group and intramyocardial injection of GFP-AD group.The mRNA levels of GFP in the heart and liver tissues were detected by Q-PCR at different time points.Fluorescence microscopy was performed to visualize the expression of GFP fluorescence.Results Compared with the tail vein injection group, the GFP mRNA level in mouse heart tissue was apparently higher in the intramyocardial injection group.In both groups, the GFP mRNA levels in liver tissue were significantly increased compared with that in the heart tissue.In the tail vein injection group, the GFP mRNA level in liver tissue reached a peak on day 7;but in the intramyocardial injection group, the mRNA level of GFP in liver tissue reached apeak on day 3.We also observed the same trend of GFP fluorescence expression in the tail vein injection group compared with that in the intramyocardial injection group.Conclusions Intramyocardial injection of adenovirus vector is suitable to achieve a higher transfection efficiency in mouse heart tissue compared with the tail vein injection method.Although both injection methods are suitable for transfection of mouse liver, the tail vein injection method is preferential for it is simple and less invasive.

Objective To evaluate the anatomic localization and size of acute necrotic myocardium in the ischemic-reperfused rat hearts using 99m TC-Glucarate and microSPECT/CT.Methods The ischemic-reperfused ( IR) rat heart models were established by ligating left anterior descending coronary artery for 60 min.99mTC-Glucarate was intravenously injected into the rats 24 hours after IR operations .Images were acquired 30 min after administration of 99m TC-Glucarate using microSPECT/CT. Anatomic localization and size of acute necrotic myocardium were analyzed with microSPECT/CT imaging , and these results were compared to those determined by triphenyltetrazolium chloride ( TTC ) staining.Results The microSPECT/CT images showed hot spot accumulations of 99mTC-Glucarate in IR hearts (the heart-to-liver ratio was 1.90 ±0.33), not in controls (P <0.05).The anatomic localization of 99mTC-Glucarate-labeled necrotic myocardium were in correspondence with TTC staining results .The hot spot size was related significantly to necrotic myocardial size determined by TTC staining ( R2 =0.964 ) .Conclusions The localization and size of acute necrotic myocardium can be assessed by non-invasive microSPECT/CT imaging with99m Tc-Glucarate.

It is very important to establish and execute the all -sided experimental animal quality monitoring in order to guarantee human health , animal health and welfare as well as the authenticity , validity, and repeatability of the experimental research results.Setting corresponding sentinel animals in the experiment can effectively monitor the quality of experimental animals.This article gives a general review of the selection of sentinel rats in the microbiological quality monitoring of the experiment animals , contact form and time between the sentinel rats and the rats being monitored , the placement of sentinel rats, and the number of rat cages being monitored by each cage of sentinel rats , as well as the test quantity, test frequency and the project.

Aim To investigate the expression of AB-CB5 and MDR1 in the cell line KG1 a and samples from acute myeloid leukemia ( AML) and their effects on multidrug resistance. Methods The expression of ABCB5 and P-gp ( the expressed product of MDR1 ) in KG1 a cells were detected by flow cytometry as well as Western blot analysis; KG1 a cells were transfected with the specific siRNA of ABCB5 using lipo2000 to reduce the expression of ABCB5; intracellular rhoda-mine123 was measured by flow cytometry;cell viability was detected by MTT; the expressions of ABCB5 and MDR1 in samples from AML were detected by real time PCR. Results ABCB5 and P-gp were overexpressed in KG1 a;the specific siRNA of ABCB5 transiently in-hibited the expression of ABCB5 in KG1 a; the siAB-CB5-KG1 a cells increased the intracellular rhodamine 123 and have been more sensitive to adriamycin com-pared with the parent KG1a. ABCB5 gene expression in samples from AML was higher than healthy people. Further, the expression of ABCB5 in 38 relapse or re-fractory AML significantly exceeded the 33 drug sensi-tive. And we found a significant positive correlation between ABCB5 expression and MDR1 gene expression in the 38 patients with relapse or refractory AML. Conclusion ABCB5 , as well as P-gp contributes to mediate multidrug resistance of AML, which provides a novel target for the therapy of relapse or refractory AML.

Objective To investigate the biomechanical and pathological changes in vivo in the taut bands (TB) of biceps femoris in rats after repeated low-frequency electrical stimulation.Methods Twenty-eight Wistar rats were randomly divided into a control group,an electrical intensity-dependent fatigue group,which were subject to electric intensity-dependent fatigue test,and an electrical frequency-dependent fatigue group,which were subject to electrical frequency-dependent fatigue test.After fatigue tests,the taut band of the biceps femoris and the non-taut band of the contralateral biceps femoris were harvested for pathological observation.The maximum contraction force (MCF),electrical intensity-and frequency-dependent fatigue characteristics and any pathological changes in the TBs were assessed and compared to the non-taut band region of the other biceps femoris.Results The MCF at the 15th and 20th stimulation (1.42 ± 0.28 g and 0.93 ± 0.54 g respectively) were significantly lower than that at the 1 st and 5th stimulation of the TBs.High stimulation intensity (HSI) at the 15th and 20th stimulation (3.76 ± 0.71 V and 3.44 ± 0.97 V) were also significantly lower than at the 1st TB stimulation.At the 10th,15th and 20th stimulation of the TBs,MCF and HSI were both significantly lower than in the bands which were not tight.In the frequency-dependent fatigue stimulation tests,the frequency which generated the MCF of the TBs was significantly lower than in the bands which were not tight,while the MCF of the TBs was significantly higher than that of non-TBs.After either intensity or frequency fatigue testing,more severe edema,uneven cytoplasmic death and degeneration of muscle fibers were observed in sections from TBs than from the bands which were not tight.Conclusions Taut muscle bands are significantly less fatigue-resistant than normal muscle fibers.Taut bands may contribute to the fatigue of myofascial pain syndromes.

Objective To investigate the effects of hyperbaric Oxygen (HBO)exposure on Caspase-3 and Bcl-2 mRNA ex-pressions in both internal carotid artery (ICA)and basal artery (BA)in rabbits.Methods Twenty-four healthy adult New Zealand rabbits were randomly divided into 2 groups:HBO group and the control group,with each group consisted of 12 animals.The rab-bits in the HBO group were exposed to HBO at 2.2 ATA for 60 minutes each day for 3 successive days.The rabbits in the control group were normally fed without any treatment.Real-time PCR was used to detect Caspase-3 and Bcl-2 mRNA expressions in both ICA and BA in 2 groups.Results HBO significantly decreased the Caspase-3 mRNA expression [(0.038 ±0.006 )vs .(1.000 ± 0.225)]and increased the Bcl-2 mRNA expression [(1.877 ±0.1 69)vs .(1.000 ±0.364)].In ICA,HBO similarly decreased the Caspase-3 mRNA expression [(0.41 9±0.091)vs .(1.000 ±0.1 75)]and increased the Bcl-2 mRNA expression [(1.269 ±0.270) vs .(1.000±0.1 1 7)]in BA.All the differences mentioned above were of statistical significance (P <0.01).Conclusion HBO ex-erts an inhibition effect on apoptosis in cerebral vascular endothelial cells.The mechanism may be related to inhibiting the expres-sion of Caspase-3 mRNA and promoting the expression of Bcl-2 mRNA.

Objective To investigate the relationship between aquaporin-1, -2, -3, -4 mRNA (AQP1-4) and renal parenchyma thickness in congenital hydronephrotic kidney in children. Methods The expressions of aquaporin 1, -2, -3, and -4 mRNA in hydronephrotic kidney of 37 children (aged 60.3±48.8 months) were evaluated with congenital hydronephrosis and control kidney of 6 children (aged 62.7±17.1 months) by using semi-quantitative reverse transcriptase polymerase chain reaction technique. Hydronephrotic kidney parenchyma thickness was measured by B-Ultrasound preoperative-ly and verified at operation. The relations of aquaporin 1, -2, -3, and -4 mRNA to the hydronephrotic kidney parenchyma thickness were analyzed by correlation analysis. Results The aquaporin 1 ,-2,-3, and -4/beta-actin ratio in the hydronephrotic kidney and normal kidney were 0.39±0.22 vs 0.90± 0.10, 0.42±0.20 vs 0.92±0.09, 0.525±0.22 vs 0.98±0.12, 0.30±0.18 vs 0.74±0.21 respec-tively, and the differences were significant (P<0.01). Hydronephrotic kidney parenchyma thickness measured by D-Ultrasound was 5.01±2.38 mm, which was identical with those measured at opera-tion. Significant correlation was found between the levels of aquaporin 1,-2,-3, and -4 mRNA and hydronephrotic kidney parenchyma thickness (r=0.773, 0.772, 0.557, 0.625, respectively; P< 0.01). Conclusions Significant correlation exists between decreased expressions of aquaporin 1 ,-2, -3, and -4 mRNA and atrophic change of renal parenchyma. This result may provide evidence to ex-plain the mechanism why the thinner renal parenchyma thickness, the weaker renal concentration and dilution function.

Objective To compare the effect of various manipulation of Tuina on surface electromyogram (sEMG) in hemiplegics after stroke. Methods From January to May, 2016, 20 inpatients with hemiplegia after stroke accepted Tuina on bilateral rectus femoris by the same therapist, with the techniques of rolling, patting, rubbing, shaking, kneading and pressing, one minute a manipulation and interval one minute. Integrated electromyography (iEMG), root mean square (RMS) and median frequency (MF) of sEMG were compared, both in rest and during Tuina. Results There was no significant difference of iEMG, RMS and MF between affected and unaffected sides in rest (t<1.147, P>0.05). iEMG and RMS were the most under patting (F>21.376, P<0.001), and MF was the highest under pressing (F>11.772, P<0.001). iEMG, RMS and MF were not very different under other manipulation (P>0.05). iEMG and RMS were less in the affected side than in the unaffected side under patting (P<0.05). Conclusion Various manipulation of Tuina may be different in neuromuscular stimulation, that patting may stimulate more muscles and motor units.