AIM: To study the suitable conditions of the extraction-separation process of the indirubin in Folium Isatidis with NIR. METHODS: The indirubin was chromatographed on an aluminium oxide column and chloroform-ethyl acetate as eluting agent,eluate was recrystalized by methanol-acetone. RESULTS: The indirubin extracted by this method was of high purity (97.78%).The sample purity was similar to reference substance one((98.01%)).The spectra of NMR and IR are the same. CONCLUSION: This method is proved to be fast,safe,lower cost,simple and has good results.

Objective To investigate the pathway of lymphatic drainage of proteins from cerebral parenchyma in subarachnoid hemorrhage rat models. Methods Healthy adult male Wistar rats were divided into Saline group, Evans blue-labeled albumin (EBA) group, and SAH + EBA group. SAH models were produced by double injection of autologous arterial blood into cisterna magna. Using a modified microinjection method, EBA was injected into left candate-putamen of the EBA group and EBA + SAH group rats. In Saline control group, saline was injected. After injection, at 12 hours, 1 day, 2 days, 3 days and 5 days, the animals were sacrificed and the fluorescence signals of EBA were imagined and analyzed along the possible lymphatic drainage pathway, e.g. the brain tissue, the wall of common carotid artery, and cervical lymphatic nodes. Results One day after injection, in EBA group, the fluorescence of EBA initially appeared on the left of the brain, the wall of common carotid artery, left lateral cerebral ventricle, and the perivascular spaces of cerebral vessels. The fluorescence signals gradually expanded to the opposite side.Large amount of fluorescence granules accumulated in the outer layer of common carotid artery. Fluorescence was also found in cervical lymphatic nodes. Two days after injection in this group, the density of fluorescencein the brain became weaker while the density of fluorescence in rhinencephalon became stronger. The fluorescence of EBA was found in lymphatic nodes adjacent to abdominal aorta. In SAH + EBA group,reduced amount and velocity of the drainage of EBA from left caudate-putamen to rhinencephalon, cervical lymphatic nodes, and lymphatic nodes adjacent to abdominal aorta were observed. From 12 hours to 5 days after injection, fluorescence intensity of EBA in deep cervical lymphatic nodes in SAH + EBA group(8.9 ±2. 0, 11.9 ± 2. 5, 17.4 ± 3.7, 26.7 ± 4. 5 and 59.0 ± 8. 1 ) were lower than those in EBA group ( 14. 5 ±3.2, 27.5 ±7.4, 60.3 ±12.3, 138.0±12.0 and 108. 1 ±13.4, F=13. 17, 24.04, 66.81, 302.77 and 59.36, P ＜ 0. 01 ). From 2 to 5 days, fluorescence intensity of EBA in lymphatic nodes adjacent to abdominal aorta was also lower in SAH + EBA group( 11.0 ± 1.5, 12. 5 ±2. 8, 23.6 ±3. 2) than those in EBA group(26. 3 ±5.9, 47.5 ±9.6, 41.0 ±9.3; F =38. 17, 72.52, 19.01, P ＜0.01). Conclusion SAH can result in reduced drainage of macromolecular substances, e.g. protein, from the brain via lymphatic pathway.

Objective To study the relationship between genetic polymorphisms of CYP2E1 and the susceptibility of the acute leukemia in Gansu population. Methods The C609T polymorphism of CYP2E1 gene was detected by polymerase chain reaction-ligase detection reaction (PCR-LDR) and 1∶1 matched casecontrol method in 100 healthy persons (control group) and 100 patients with acute leukemia (AL group).Results The C2 allele genotype and C1C2/C2C2 genotype of CYP2E1 gene occurred more frequently in AL group (13.5 % and 22 %, respectively) than those in control group (10.5 % and 19 %, respectively), however,both differences showed no statistical significant. Further stratified analysis, the C1C2/C2C2 genotype of CYP2E1 gene occurred more frequently in AML group (27%) than that in control group (19 %), but difference had no statistical significant, too. The occurrence frequency of the C2 allele genotype and C1C2/C2C2 genotype of CYP2E1 gene showed no significant difference in ALL group and control group (x2=0.446, P =0.504>0.05). Conclusion Genetic polymorphisms of CYP2E1 don't correlated to susceptibility of acute leukemia(AML and ALL) in Gansu population.

AIM:To determine the injured effect of cerebrospinal fluid(CSF) from subarachnoid hemorrhage(SAH) after cerebral lymphatic blockage(CLB) on PC12 cells. METHODS:SAH and CLB models of adult New Zealand rabbits were used. CSF was obtained from experimental animals after 5 d of modeling and was added into cultured PC12 cells. The cells were randomly divided into blank control(F12 Ham's),normal CSF,SAH CSF,and SAH+CLB CSF groups. At different time points,the survival rate of PC12 cells was measured by MTT assay. LDH leakage was detected. Expression of Bax and heat-shock protein 70(HSP70) was determined by immunohistochemical staining. RESULTS:MTT assay and detection of LDH leakage revealed that the survival rate of PC12 cells was obviously inhibited and the leakage of LDH increased in SAH CSF group and SAH+CLB CSF group. CSF from normal rabbit did not damage the PC12,as compared to blank controls. Above effects were more obvious in SAH+CLB CSF group than those in SAH CSF group. Bax and HSP70 protein expression was found in both SAH CSF group and SAH+CLB CSF group. Expression of Bax protein in SAH+CLB CSF group was stronger than that in SAH CSF group in a time dependent manner. At 0.5 h and 1 h,the expression of HSP70 protein in SAH+CLB CSF group was stronger than that in SAH CSF group,whereas the expression became weaker at 2 h and 4 h in that group. CONCLUSION:Blockage of cerebral lymphatic drainage pathway deteriorates the damage of CSF from SAH on PC12 cells,indicating this pathway may acts as an endogenous protective mechanism in SAH.

Aim To establish a method to evaluate lymphatic drainage of macromolecular tracer in cerebrospinal fluid in rats.Methods Rat cervical lymphatic blockade(CLB)models were established by occlusion of cervical lymphatic tubes and removal of cervical lymphatic nodes.The rats were divided into non CLB(normal controls) and CLB groups.~(125)I-labeled human serum albumin(~(125)I-HSA)was injected into the left lateral cerebral ventricle,and blood samples were collected and ~(125)I-HSA concentrations were detected continually within 24 hours.Concentration-time curve was drawn according to the single compartment model in pharmacokinetics.Parameters of pharmacokinetics such as area under curve(AUC),maximum concentration(C_(max)),transfer rate constant K_a and peak time(T_(max))were derived.The AUC,C_(max),K_a,and T_(max) regarding the lymphatic drainage of ~(125)I-HSA were calculated based on the differences between the two groups.Results AUC,C_(max),K_a of ~(125)I-HSA by lymphatic drainage were 51.97 mg·L~(-1)·h~(-1),2.91 mg·L~(-1),and 0.64 h~(-1),respectively.The proportion of AUC,C_(max),K_a of ~(125)I-HSA by lymphatic drainage to those of drained by both arachnoid granulations and lymphatics was 71.53%,44.02%,58.18%,respectively.T_(max) in CLB group(8.36±0.82 h)was much longer than that in non CLB group(3.57±0.54 h).Conclusions A method to evaluate lymphatic drainage of macromolecular tracer in cerebrospinal fluid in rats is successfully established.The lymphatic drainage pathway plays an important role in eliminating macromolecular substances in cerebrospinal fluid.

Objective To investigate the influence of cerebral lymphatic blockade (CLB) on apoptosis of hippocampal neurons after subarachnoid hemorrhage (SAH) in rats. Methods Healthy adult Wistar rats were randomly assigned to normal control group,SAH group and SAH + CLB group. SAH model was induced by double injection of autologous blood into the cistema magna. On day 3 after second injection, hippocampal cell shape structure of each group were determined by hematoxylin-eosin staining (HE) and propidium iodide (PI) staining. Terminal-deoxynucleotidy transferase mediated nick end labeling (TUNEL) fluorescent was used to determine the situ apoptosis. Immunohistochemistry was conducted to study the expression of caspase-3 and Bcl-2 in hippocampal neurons. Results (1) HE staining and PI staining showed the hippocampal neurons of SAH rats were partly shrink,and nuclei showed wavy or folded seam-like,some crescent-shaped; the hippocampal neurons in SAH + CLB group distributed sparsely,nuclear fragmentation,apoptotic bodies could be seen,surrounded by vacuole formation, Compared with the SAH group, the number of apoptotic cells in SAH + CLB group was significantly increased(the number of apoptotic cells: 0.71 ±0.05,25.36 ±4. 02,37. 82 ±5.93, P<0.01). (2) The fluorescence intensity of positive cells by TUNEL stain in SAH group and SAH + CLB group was higher than in normal control group,while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.19 ±0.03,1.70 ±0.37,2.54±0.53, P<0.01). (3) The fluorescence intensity of caspase-3 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.14 ±0.03,2.45 ±0.49,2.96 ±0.44, P<0.01). (4) The fluorescence intensity of Bcl-2 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly lower than the SAH group(the fluorescence intensity: 0.58 ±0.08, 3.40 ±0.61,2.67 ±0.44, P<0.01). Conclusion Cerebral lymphatic blockade induce the apoptosis of hipp-ocampal neurons in rats after SAH,which mechanism may be related to high expression of caspase-3 and low ex-pression of Bcl-2.

Objective Our retrospective study was aimed to analyze the clinical value of serum CEA and CA19-9 in patients with chole-lithiasis.Methods The clinical data of 98 patients with cholelithiasis and 44 patients with inguinal hernia received treatment in our hospital from February 2011 to February 2015 were retrospectively analyzed.The expressive levels of CEA and CA19-9 of the patients were detected and compared.The important roles of CEA and CA19-9 in the patients with cholelithiasis were analyzed.Results The levels of CEA,CA19-9 and inflammatory factors in normal group and control group had no statistical differences (P>0.05).The levels of CEA,CA19-9 and inflam-matory factors in rising group were higher than those in control group (P<0.05).The levels of CEA,CA19-9,inflammatory factors before and after the treatment had no statistical differences (P>0.05),but the levels of CEA,CA19-9,inflammatory factors in rising group were obvi-ously decreased( P<0.05) .Conclusion The levels of CEA and CA19-9 in patients with cholelithiasis had correlation with inflammation of biliary tract,which will increased by severe choleithiasis.

Objective To prepare Wurenchun solid dispersion of alcohol extracts of Fructus Schisandrae Chinensis so as to improve the dissolution of its active compound in vitro.Methods The Wurenchun solid dispersions were prepared with various carriers and drug/carrier ratios by mixing the carrier in alcohol extractive solution of FSC directly,and the apparent solubility and dissolution of deoxyschisandrin in them were tested and compared.Results The apparent solubility and dissolution of deoxyschisandrin of Wurenchun solid dispersion(extracts: polyvinylpyrrolidone(PVP) K30=1∶3) were increased remarkably to 5.06 ?g/mL and 43.2% in water individually,including dispersed and dissolved drug whose particle size is below 0.22 ?m,compared with that of the self-prepared Wurenchun capsules.Conclusion Wurenchun solid dispersion made of PVP K30 can remarkably enhance apparent solubility and dissolution of the active compound in vitro.