To establish a stable method for determining the tannins in Rosa laevigata Michx. by casein method. The optimum conditions of determination were adopting 400mg caseins,adding 1.5% Na2CO3 and detemining after 30 min. The selected method has a good liner dependance. The average recovery of tannin acid was 100.5%,RSD=1.85%. The method is rapid and reliable for the determination of tannins in Rosa laevigata Michx. and determined the contents of the fruits in eight different disturbs and xi’an 's content has the highest,3.96%.

Objective: To study the method of identification and content determination of the important component in TongRenNiuHuangQingXin Tablet.Method:The major compounds in this drug were identifi ed by the thin layer chromatographic method and the contents of paeoniflorin were determined by HPLC.The HPLC procedure was used in the determination: the chromatographic column was Symmetry ShieldTM Rp18(3.9mm?150mm,5?m),the mobile phase was methanol-0.05mol/L potassium dihydrogen phosphate(25:75) with a flow velocity of 0.8ml/min,the detection wave length was 230nm,and the room temperature was 30℃.Results: Calculus Bovis Artifactus and Gensing were identified.The regression equation of paeoniflorin was in the range of 0.10-0.51?g.The average recovery of the procedure was 98.94%(RSD=2.05%).Conclusion: The identifi cation and determination methods established were specifi c and repeatable,which can be used to control the quality of TongRenNiuHuangQingXin Tablet.

Objective:To observe the effects of Shen Hong Bu Xue Granule(SHBXG) on the expression levels of erythropoietin(EPO)mRNA in kidney tissue and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA in bone marrow tissue of the mice with blood deficiency,and to investigate the protective effect of SHBXG on the blood deficiency mice and its mechanism.Methods:The mouse model of blood deficiency was established with acetylphenylhydrazine(APH) and cytoxan(CTX).A total of 60 mice were divided into blank control group,model group,Compound E-jiao Slurry group and low,middle,high doses of SHBXG groups(n=10).The serum,kidney and bone marrow tissues from all the mice were collected after 14 d consecutive administration.The levels of red blood cells(RBC),hemoglobin level(HGB),hematocrit(HCT),leukocytes(WBC),and platelet(PLT) in blood of the mice were detected by automatic blood analyzer;the levels of serum EPO and GM-CSF of the mice in various groups mice were detected by ELISA method;the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice were detected by RT-PCR method.Results:The results of automatic blood analyzer showed that the levels of RBC,HGB,HCT,and PLT of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group (P<0.05 or P<0.01);the levels of WBC of mice in Compound E-jiao Slurry group and high dose of SHBXG group were increased significantly compared with model group(P<0.05).The ELISA results showed that the levels of serum EPO and GM-CSF of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group(P<0.05).The PC-PCR results showed that the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice in Compound E-jiao Slurry group and middle and high doses of SHBXG groups were increased significantly compared with model group(P<0.05 or P<0.01).Conclusion:SHBXG could improve the blood deficiency symptom in the mice with blood deficiency,and its mechanism may be related to increasing the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice.

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, which provides a new therapeutic idea for myocardial injury.OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P <0.05) and its membrane potential decreased significantly compared with the blank control group (P < 0.05). Theapoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantlycompared with the 5-azacytidine group (P < 0.05, P < 0.01), and their membrane potential increased significantly (P <0.05, P < 0.01). The results of western blot showed that the expression level of Nkx2.5 increased significantly in thepilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while theexpression levels of GATA4 and ATF-2 in each group were low and there were no significant differences among groups(P > 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4,ATF-2 and MEF-2C needs to be further studied.

OBJECTIVE:To establish the HPLC fingerprint for Anemone raddeana. METHODS:HPLC was performed on the column of Phemomenex Gemini C18 with mobile phase of 0.1%phosphoric acid-acetonitrile(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 206 nm,the column temperature was 30℃,and the injection volume was 20μl. With the refer-ence of raddeanin A,13 batches of A. raddeana were analyzed,chromatographic fingerprint similarity evaluation system software was conducted for similarity analysis,and SPSS 13.0 was conducted for cluster analysis. RESULTS:There were 11 common peaks in the 13 batches of A. raddeana with similarity of higher than 0.90. According to the verification,the fingerprint and control fin-gerprint shows good consistency. The drugs in Huadian,Jiaohe, Tiangang,Shulan,Tonghua and Fusong of Jilin and Shangzhi of Heilongjiang were regarded as category 1,and in Harbin,Yabuli town and Yimianpo of Heilongjiang,Qingyuan of Liaoning,Ji-nan of Shandong were category 2. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of A. raddeana.

AIM: To study the application of near infrared spectroscopy for quality control of in-process Wuji Baifeng Pill(Gallus Domesticus,Colla Cornus cervi,Carapax Trionycis,Otheca Mantidis,Radix et Rhizoma Ginseng,etc.). METHODS: Qualitative and quantitative methods for analysis of in-process product was developed by diffuse reflectance near infrared spectroscopy combined with chemometrics.Spectra of 95 samples of Tongren Wuji Baifeng Pill in-process and 19 raw material negative in-process products were collected.Similarity match model was built on the basis of spectra of 75 batches of qualified in-process products.Using this model,the similarity match values of other samples were calculated.Multivariate calibration models for paeonin and water content in in-process product were developed with partial least square algorithm. RESULTS: The results showed that the proposed similarity match model could reflect the quality characteristic of Tongren Wuji Baifeng Pills in-process and reveal the absence and presence of raw materials.The correlation coefficients of the calibration model were within 0.965 7 and 0.992 1 while the values of root mean square error of prediction were 0.005 5% and 0.45%,respectively. CONCLUSION: The experiment shows that the established method is accurate,fast and non-destructive,and can be applied to on-line detection.

Objective To investigate MRI features of heat stroke (HS) and 1H-MRS in monitoring clinical outcome.Methods Six cases(4 males,2 females) of HS patients,aged 43-75 years old were retrospectively studied.The patients were divided into 4 severe cases (2 cases with coma for 4 hours,2 patients with persistent coma) and 2 moderate cases according to initial Glasgow coma scale (GCS) at the time of hospital admission.All patients underwent T1WI,T2WI,FLAIR,DWI,SWI,multi voxel 1H-MRS scans and the ratios of NAA/Cr,Cho/Cr,and NAA/Cho were calculated.One case of severe coma died on the next day.Follow-up imaging and brain 1H-MRS were performed on other 5 patients on the 7th and 15th day after the first examination.The imaging results were compared with clinical outcome.Results MRI features of HS for severe coma patients were iso-intensity on T1WI,hyper-intensity on T2WI,FLAIR and DWI,heterogeneous signal intensity on ADC.The infratentorial lesions distributed in the cerebellum,dentate nucleus and cerebellar arm (3 cases),and the supratentorial lesions distributed in the parahippocampal gyrus (2 cases),parietal lobe (2 cases),and temporal lobe (1 cases).The lesions could be multiple,and the cerebellum and the parahippocampal gyrus were most common involved.SWI showed focal hemorrhage in 2 cases,which located in the brainstem and parietal lobe.No obvious MRI abnormality was found in 2 cases of moderate coma patients.The ratios of NAA/Cr,Cho/Cr,and NAA/Cho for severe coma patients were 0.67 ±0.09,0.94 ±0.16 and 0.70±0.12,respectively,which for moderate coma patients were 0.87±0.12,0.95±0.13,and 0.94±0.08,respectively.The NAA/Cr and NAA/Cho ratios decreased and Cho/Cr mild decreased in the severe coma patients when compared with the moderate coma patients.One patient with persistent coma had expanded lesions on follow-up study.NAA/Cr,NAA/Cho and Cho/Cr were 0.63± 0.07,0.67 ±0.10,and 0.96 ±0.05,respectively.Two cases had smaller lesions on follow-up study with NAA/Cr and NAA/Cho ratio increased,which were 1.02±0.13 and 0.96±0.11,respectively.GCS was turned to 14.NAA/Cr and NAA/Cho for moderate coma patients were 1.17±0.10 and 1.21 ±0.07,respectively.Conclusions Severe HS patients present with cerebral edema and hemorrhage.1H-MRS can be an important index for monitoring the severity of HS.The longer the duration of coma,the worse prognosis may happen.

To observe the effects of emodin ( Emo) on acute fatty liver in mice induced by DL-ethionine ( DL-Eth) or tetracyclin ( Tetra) and its potential mechanism, ICR mice of acute fatty liver model induced by DL-Eth were orally administered with Emo or positive control, ursodeoxycholic acid ( UDCA) for 7 days. On day 7, except that the control and Emo groups were treated with vehicle control, animals were orally administered with DL-Eth to induce acute fatty liver model. ICR mice of acute fatty liver model induced by Tetra were orally administered with Emo or positive control, Dong Bao Gan Tai ( DB) or total flavonoid C-glycosides from Abrus mollis extract ( AME) for 7 days. From day 4, except that the control group was treated with vehicle control, animals were injec-ted with Tetra intraperitoneally for 4 days to induce acute fatty liver model. Liver histopathological analyses were determined by HE staining. Serum aspartate transaminase ( AST) , alanine transaminase ( ALT) , serum triglyceride ( TG) , hepatic TG and hepatic total cholesteol ( TC) were detected. The expression of phosphorylated AMP-activa-ted kinase ( p-AMPK) , phosphorylated acetyl-CoA carboxylase ( p-ACC) , SREBP1 and fatty acid synthase ( FAS) were determined by Western blot. The expression of fatty acid translocase ( CD36 ) , peroxisome proliferator activa-ted receptor alpha ( PPARα) and microsomal triglyceride transfer protein ( MTTP ) in liver were determined by RT-PCR. Compared with model groups, Emo could improve hepatocyte swelling and hepatic steatosis induced by DL-Eth or Tetra. Serum AST, ALT, serum TG, hepatic TG and hepatic TC were decreased by Emo significantly. DL-Eth-induced increase of fatty acid synthetase-associated protein was down-regulated by Emo. Fatty acid uptake was down-regulated by Emo; fatty acid oxidation and secretion were increased by Emo. Emo might be effective in preventing acute fatty liver in mice induced by DL-Eth or Tetra.

Objective To evaluate the clinical value of susceptibility?weighted imaging(SWI) in detecting intramedullary hemorrhage of traumatic acute spinal cord injury. Methods From October 2012 to December 2014, 37 TSCI (traumatic spinal cord injuries) patients undergone the MRI scans including routine MRI and SWI were enrolled. Further according to ASIA classification standard, all patients were evaluated as ASIA A (n=4), ASIA B (n=4), ASIA C (n=19) and ASIA D (n=10). Referring to axial T2WI images at the same slice, the manifestations of hemorrhage in amplitude image, phase image and SWI were evaluated. At the slice with maximal size of hemorrhage area and its neighboring slices, the hemorrhage regions were manually drawn; and the total area was automatically calculated. The number of hemorrhage lesions was defined as the number of hemorrhage lesions at single slice × slice number. One?way ANOVA was used to compare the differences among different grading ASIA in terms of hemorrhage area and number. Meanwhile, the relations between hemorrhage area and ASIA grade; hemorrhage number and ASIA grade were evaluated by Spearman rank correlation. Results The hemorrhage was detected by SWI in 15 patients, including 4 ASIA A, 4 ASIA B, 7 ASIA C. Hemorrhage represents as isointense in T1WI and a slightly low signal intensity or isointense in the center companied by high intensity at circus in T2WI. In magnitude image and SWI hemorrhage appears as low signal intensity and low signal intensity in the center companied by high intensity at circus in phase image. In detecting the hemorrhage, SWI (98 lesions were detected) was 5.4 times of T2WI (19 lesion were detected). As for the number of hemorrhage, significant differences were found among ASIA grading A, B and C (grading A:22.5 ± 1.3, grading B:19.5 ± 1.3, grading C:4.0 ± 1.1;F=38.720, P<0.01); Further the pairwise comparison showed statistical significance (P<0.05); besides, the number of hemorrhage lesions closely related with the ASIA grade (r=0.864, P<0.01). For the hemorrhage area, the calculated results of grading ASIA A, B and C were (23.5 ± 0.6), (21.8 ± 1.9), (3.9 ± 0.7) mm2, respectively; there were significant differences among the different ASIA grades (F=29.987, P<0.001);furthermore the hemorrhage area also showed closely relation with the ASIA grade (r=0.778, P<0.01). Conclusions SWI is more sensitive in detecting the hemorrhage in traumatic acute spinal cord injury. The more number and area of bleeding area suggest the more severe of the damage level.

Objective To introduce the application of computer aided technology in the treatment of pelvic malunion and observe the effect of this technique on the operation.Methods Data of 9 consecutive patients with pelvic malunion who were enrolled in our hospital from January 2009 to December 2012 were retrospectively observed.There were 4 patients who had undergone surgery with computer aided technology,including 3 men and 1 woman with an average age of 34 years (computer aided technology,CAT) group.According to the Tile classification,4 cases were all type C.There were 1 case with open pelvic fractures,2 associated with the craniocerebral injury,2 associated with the thoracic injury and 2 associated with the abdomen trauma.There were 5 patients who had undergone surgery without computer aided technology,including 3 men and 2 women with an average age of 32.6 years (conventional group).According to the Tile classification,4 cases were all type C.There were 1 case with open pelvic fractures,2 associated with the craniocerebral injury,3 associated with the thoracic injury and 1 associated with the abdomen trauma.The Majeed score,the visual analogue score (VAS),operation time,blood loss,blood transfusion,intraoperative fluoroscopy,degree of improvement in lower limb length,iatrogenic injury and the complication were all recorded respectively.Results For the CAT group,the operation time was 195-230 min,the blood loss was 800-1 6 00 ml,the blood transfusion was 6-16 U,intraoperative fluoroscopy was 6-11 times and the degree of improvement in lower limb length was 2-3 cm.The Majeed score in the final follow-up was 78-90 points,including 3 excellent cases and 1 good.The VAS after surgery was 0-4 points.For the conventional group,the operation time was 210-330 min,the blood loss was 600-4 500 ml,the blood transfusion was 6-28 U,intraoperative fluoroscopy was 7-18 times and the degree of improvement in lower limb length was 1-3 cm.The Majeed score in the final follow-up was 79-89 points,including 3 excellent cases and 2 good.The VAS after surgery was 1-4 points.Conclusion Surgeons can make full preoperative planning by the computer aided technology before the operation.This technology which reduces the operation time can make the operation more accurately,effectively and safely.