Objective To study the best extraction method for Radix Salvia Miltiorrhiza effective component. Methods Several extraction methods for Radix Salvia Miltiorrhiza (water decoction, alcohol reflux, ultrasonic extraction and supercritical CO2 extraction) were compared, to optimize the extraction methods by determined of Danshensu and Tanshinone IIA of Radix Salvia Miltiorrhiza. Results Supercritical CO2 extraction had high content of effective component of Danshensu and Tanshinone IIA. Conclusion Supercritical CO2 extraction was the best extraction method for Radix Salvia Miltiorrhiza. This result would be an experimental proof for pre-processing the preparation of Radix Salvia Miltiorrhiza.

AIM:To observe the relation between astroglial activation state and ischemic tolerance in the gerbil hippocampus. METHOD: Bilateral occlusion of common carotid arteries and immunofluorescent methods in the gerbil hippocampal tissue. Slices were used. The morphological changes of the neurons were observed by light microscopy.RESULTS:Pretreatment with 2 minute bilateral carotid artery occlusion produced protective effects of CA 1 neurons. Glial fibrillary acidic protein (GFAP) staining showed weak positive cells in control group. Most of GFAP positive cells showed weak and middle positive cells after recirculation 1d and 2d following ischemic 3.5 min and preconditioning of the brain with sublethal ischemia respectively.CONCLUSION:Astroglial played an important role in cerebral ischemia. It is possible that state of astroglial activation related to neuronal survival in ischemic tolerance.

AIM To observe the effects of L NNA on glial fibrillary acidic protein (GFAP) production in the gerbil hippocampus after recirculation following ischemia. METHODS Both vessels occlusion and immunofluorescent methods in gerbil hippocampal tissue slices incubated in vitro were used. RESULTS Recirculation following ischemia led to a rise on GFAP concentration in the hippocampus. GFAP positive cells mainly distributed in rediatum layer and molecullar layer in the hippocampus, L NNA decreased GFAP synthesis. CONCLUSION L NNA is a strong inhibitor of nitric oxide synthase(NOS). It is possible that the role of endogenous NO mediated GFAP synthesis is inhibited by L NNA.

Objective To investigate the effect of miR-141 up-regulation through miR-141 mimic transfection on proliferation,apoptosis and cell cycle of hepatocellular carcinoma (HCC) cells.Methods Real-time polymerase chain reaction (qPCR) was employed to detect differential expression of miR-141 in normal hepatic epithelial cells (L02) and hepatocellular carcinoma cells (HepG2).Furthermore,miR-141 mimics were used to transfect into HepG2 to up-regulate the expression of niR-141 (experimental group).Blank group (CON) and negative control group (miR-NC) were used as control group.qPCR was used to detect expression of miR-141.CCK8 assay was used to detect the proliferation of HepG2 cells.Flow cytometry was used to detect the apoptosis and cell cycle of HepG2 cells.Results The results of qPCR showed that miR-141 was significantly down-regulated in HepG2 cells (0.64 ± 0.13) compared to L02 cells (1.00 ± 0.18),and the difference was significant (P ＜ 0.05).Relative expression of miR-141 was significantly increased after transfection (3.33 ± 0.66),compared to CON group (1.00 ± 0.17,P ＜ 0.05) and miR-NC group (1.08 ±0.14,P ＜0.05).CCK8 assay showed that the absorbance values of HepG2 of miR141 group at 48,72,96 h (0.67 ± 0.07,1.17 ± 0.05,1.36 ± 0.03) were all decreased significantly compared with those in CON group (0.81 ±0.02;1.42±0.03;1.73 ±0.05,all P＜0.05) and miR-NC group (0.78 ±0.01;1.38 ±0.02;1.69 ±0.01,all P ＜0.05).The results showed that miR-141 group [(11.81 ± 0.23)％] had significantly increased apoptosis rate compared to CON group [(4.18 ± 0.18) ％] and miR-NC group [(4.04 ± 0.08) ％],and the difference was statistically significant (P ＜ 0.05).miR-141 group had decreased percentage of S phase cells [(19.89 ± 2.78) ％] compared to CON group [(31.87 ± 1.00) ％,P ＜ 0.05] and miR-NC group [(30.49 ± 1.73) ％,P ＜ 0.05],while miR-141 group had significantly increased percentage of G1 phase cells [(74.74 ±2.03)％] compared to CON group (60.85 ± 1.69) ％ and miR-NC group (60.93 ± 1.95) ％,and the differences were all statistically significant (all P ＜ 0.05).Conclusions miR-141 was in low expression in HepG2 cells.Upregulation of miR-141 could specifically inhibit proliferation and promote apoptosis of HepG2 and change the distribution of cell cycle.

Objective To explore the clinical effects of injecting methotrexate with hysteroscopic tubal can-nulation combined with Huayu shapei prescription in the treatment of salpingocyesis.Methods 82 unruptured salpin-gocyesis patients were randomly divided into the two groups.41 patients in the observation group were treated by hyst-eroscopic tubal cannulation injection of methotrexate combined with Huayu shapei prescription,41 patients in the con-trol group were treated by methotrexate intramuscular injection with oral mifepristone.The clinical efficacy of the two groups were observed,the level of serumβHCG required for tubal pregnancy,the mass absorption and after treatment of tubal patency.Results Compared with the control group,the difference was significant of the decline of serum βHCG in the treatment group (t=2.67,P<0.05).Compared with the control group,the treatment effectiveness and the absorption of the mass in fallopian tube in the treatment group,the difference was significant (t =3.57,P<0.05 ).Tubal patency detected treated group was superior compared with the control group,the difference was signifi-cant(χ2 =100.23,P<0.05).Conclusion Compared with traditional treatments,injecting methotrexate with hyste-roscopic tubal cannulation combined with Huayu shapei prescription in the treatment of salpingocyesis can effectively reduce the HCG levels,promote the absorption and softening of the ectopic pregnancy mass,to achieve the purpose of killing embryos eliminate micro,it will help to improve patients with clinical symptoms and signs,to maximize the recovery of the patient's reproductive function and physical health.

OBJECTIVE：To observe effect of L-NAME on apoptosis in the gerbil hippocampus after recirculation following ischemia.METHOD：Bilateral occlusion of common carotid arteries and TUNEL stain methods in gerbitil hippocampal tissue slice were used.RESULTS：L-NAME inhibited apoptosis in the CA1 subfield.CONCLUSION:Endogenous NO mediated apoptosis is inhibited by the L-NAME.

BACKGROUND:A variety of embryonic stem cells induction and differentiation systems have been established so far, while the research that promotes embryonic stem cells to differentiate into hematopoietic stem cells is stil at an initial stage, and the induction efficiency needs to be improved.
OBJECTIVE:To active the Wnt/β-catenin signal pathway in mouse embryonic stem cells with exogenous win3a as an inducer, and then to observe whether the activation of this pathway wil promote the directional differentiation of embryonic stem cells into hematopoietic progenitor cells.
METHODS:The ES-E14TG2a mouse stem cells were cultured with the exogenous wnt3a (100 μg/L) for 21 days, the content ofβ-catenin was tested by cellimmunofluorescence and western blot, and expression of Wnt downstream target gene was detected by quantitative reverse transcription PCR to determine the activation of Wnt/β-catenin signal pathway. Single-layer adherent culture method was used to induce the directional differentiate of above-mentioned cells into hematopoietic stem cells, and detection of hematopoietic development associated surface marker CD34+/Sca-1+was achieved by flow cytometry;meanwhile, the expression of hematopoietic associated gene was measured by quantitative reverse transcription PCR.
RESULTS AND CONCLUSION:We found thatβ-catenin accumulated in ES-E14TG2a mouse stem cells after cultured with wnt3a (100 μg/L) for 21 days;the expressions of Wnt downstream target genes such as Pitx2, Frizzled, Sox17 and Oct4 showed the different degrees in increase, meaning the activation of Wnt/β-catenin signal pathway. Furthermore, during the time that we used single-layer adherent culture method to induce hematopoietic stem cells, the CD34+/Sca-1+cells accounted for 20.2%of total cells at day 14, and control cells only accounted for 11.9%. Again, expression quantity of hematopoietic associated gene BMP4, FLK2 and CD34 increased while Smad5 was suppressed significantly. Our data suggest that sustaining action by wnt3a wil active Wnt/β-catenin signal pathway, and also promote the directional differentiation of ES-E14TG2a mouse stem cells into hematopoietic progenitor cells.

Objective To explore the imaging features of bone marrow edema(BME)and soft tissue edema(STE)caused by bone tumors and tumor-like lesions.Methods Ninety nine patients with bone tumors and tumor-like lesions which were proved by surgical pathology were reviewed.The patients were divided into benign and malignant groups.Evaluation parameters included the size and signal intensity of BME and STE,the features of enhancement,the bone sclerosis and its relation with BME,and joint effusion,et al.The data of two groups were analyzed by X2 test.Results There were 40 patients in benign group and 59 patients belonged to malignant group.BME and STE demonstrated low signal on T1-weighted images and high signal on fat-suppressed T2-weighted images.Some BMEs demonstrated low signal on T2weighted images,which corresponded to sclerosis on X-ray film and(or)CT.Both BME and STE demonstrated uniform enhancement.There were statistically significant differences between benign and malignant groups including the frequency of BME.sclerosis.the median of the size of the BME and STE (P<0.05),which were 57.5%(23/40),25.0%(10/40),2.7 cm,1.3 cm,and 32.2%(19/59).3.4%(2/59),1.6 cm,1.7 cm respectively.No significant differences between the frequency of STE,joint effusion in 2 groups(P>0.05).Conclusions Both benign bone tumors and tumor-like lesions and malignant bone tumors can be accompanied by BME and STE.The size of BME in the benign bone tumors and tumor-like lesions is bigger than those in malignant ones,and the size of STE in malignant bone tumors is bigger than benign ones.

Objective To evaluate the surgical treatment of splitting right hepatic duct with hepatolithiasis and stenosis. Methods The clinical data of 38 patients with splitting right hepatic duct and hepatolithiasis treated by operation were analyzed retrospectively. Results All the patients underwent operation. operative procedures were as follows: (1) in situ cholangioplasty of splitting right hepatic duct in 7 cases;(2) fenestration of splitting right hepatic with adjacent hepatic duct in 9 cases; (3) bilioplasty of splitting right hepatic duct with adjacent bile duct in 8 cases; (4) hepatic lobectomy or segmentectomy of splitting right hepatic duct in 14 cases. Postoperative complications developed in 6 cases, which were cured conservatively. There was no perioperative mortality. All patients were followed up for 5～16 years(averaged 9.2 years). Excellent rate was 78.9%,and residual stones were found in 26.3% of the patients . Conclusions Accurate localization and appropriate operation may get satisfactory result in treating patients with splitting right hepatic duct with hepatolithiasis and stenosis.

Objective To evaluate the procedure of surgical treatment of intrahepatic biliary calculi with blood vessel variation in hepatic hilus. Method The clinical data of 57 patients with intrahepatic biliary calculi and blood vessel variation in hepatic hilus treated by biliary operation were retrospectively analyzed.Results The simply biliary operation was performed on 13 cases ; variant blood vessels were cut off and ligated in 14 cases;The bile duct and blood vessel across conversion operation were performed on 22 cases;and the intrahepatic cholangiojejunostomy was carried out in 8 cases. There were no severe postoperative complications and perioperative mortality.51 patients(89.6%) were followed up for 4 to 15 years with the exellent results in 43(84.3%)cases. There were residual stones in 7 cases,of them,5 patients had intermittent abdominal pain in the right upper quadrant; reoperation was performed on another 2 cases because of recurrent cholangitis. One died of variceal bleeding 1.5 years after operation. Conclusions In order to accomplish the complex operations of biliary tract and avoid ischemic damage of liver, proper operation should be chosen for patients with intrahepatic biliary calculi with blood vessel variation in hepatic hilus.