Objective To observe the effect of Chinese medicine of replenishing the liver and kidney on the concentration of 5-hydroxytryptamine(5-HT)in hippocampus and the expression of tryptophan hydroxylase 2 (TPH2)in median raphe nuclei in perimenopausal depression model rats and to explore its mechanism.Methods The animal model was established by resecting the ovaries of female rats,then giving isolation-housing in combination with 21 days of chronic unexpected mild stress(CUMS).High performance liquid chromatogram-electrochemical detection(HPLC-ECD)were employed to measure 5-HT in hippocampus,the hemi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)and fluorescence immunohistochemistry were employed to detect the relative expression of TPH2 mRNA and number of TPH2 immunoreactive neurons in median raphe nuclei.Results The concentration of 5-HT in hippocampus was increased in rats of the medicine group compared with that in the model group((2543.06±859.59)ng/g tissue,(1845.81±233.55)ng/g tissue,F=9.617,P＞0.05).RT-PCR showed that the relative expression of TPH2 mRNA in medicine group was much higher than that in model group ((1.282±0.158),(0.985±0.120),F=3.552,P＜0.05).Immunofluorescence showed that the number of TPH2 immunoreaetive neurons in raphe nuclei had statistical significance between medicine group and model group ((152.74±68.52),(74.12±38.01),F=7.939,P＜0.01).Conclusion Chinese medicine replenishing liver and kidney have effects of improving perimenopausal depression,increasing the expression of TPH2 in median raphe nuclei,which leads to enhance the synthesis of 5-HT in hippocampus might be one of its mechanisms.

AIM: To make evaluation on fluid resuscitation with either hypertonic saline(HS) or dextran 40(Dx) on pancreatic microthrombi and dysfunction of microcirculation in rats with severe acute pancreatitis(SAP).METHODS: SD rats were allocated into 4 groups randomly,ie.SAP group,HS group,Dx group,which respectively received normal saline(NS,4 mL/kg),HS(4 mL/kg),Dx(4 mL/kg) for 2 h by the tail intravenous injection consecutively after being made as SAP animal models,and operate sham group(OS).12 h after the operation,all animals were blooded to assess the serum amylase levels,plasma D-dimer,von Willebrand factor and GMP-140 levels.The amount of ascites was measured and the samples of the pancreas were collected for pathologic examination under light microscopy as well as transmission electron microscope.The numbers of pancreatic microthrombi were also counted with microscopy.RESULTS:(1) 12 h later when the rats were sacrificed,the survival rate in SAP group was the lowest,significantly lower than that in the 2 fluid resuscitation groups(P

Double cannulation model of conscious rat allowing simultaneous collection of mesenteric lymph and jugular venous blood was established to investigate the intestinal lymphatic transport of breviscapine orally administered in rat. The concentrations of breviscapine in plasma and lymph were determined by HPLC. The pharmacokinetics of breviscapine after oral and intravenous administration was evaluated in the conscious rat model. It was observed that scutellarin distributed from blood circulation to lymphatic system after intravenous injection. The cumulative lymphatic transport amount within 12 h was (2.78 +/- 0.25) microg, equivalent to 0.0792% of intravenous dose. After oral administration of scutellarin to double-cannulation rats, the cumulative lymphatic transport amount within 12 h was (0.92 +/- 0.08) microg, equal to 0.0083% of oral dose. The absolute bioavailability of breviscapine orally administered to double-cannulation rats was 4.91%, indicating that scutellarin was mainly absorbed into the bloodstream through the portal vein. Lymphatic transport of scutellarin appears to reflect high affinity for the lymph lipoproteins to chylomicron. This study provided a biopharmaceutics basis for developing oral lipid delivery system for the promotion of intestinal lymphatic transport to improve oral bioavailability of breviscapine.

Objective To investigate the protective effect of lipsomal clodronate against hepatic injury in rats with acute necrotizing pancreatitis (ANP). Methods 48 SD rats were randomly divided into control group, ANP group and lipsomal clodronate group, respectively. The models of ANP were established by injection of sodium taurocholate into the pancreatic capsule. Lipsomal clodronate was prepared by means of thin film. Blank liposomes and clodronate-containing liposomes was injected via caudal vein in ANP group and lipsomal clodronate group, respectively. The rats were sacrificed at 2, 6 h after ANP induction, the serum levels of ALT, AST and AMS, IL-6,IL-12 were measured, and pathologic changes of liver and pancreas were observed. Results At 6 h, serum level of ALT was (73 ± 11) U/L, (257 ± 33) U/L and (184 ± 29) U/L in control group, ANP group and lipsomal clodronate group, respectively;serum levels of AST were (190 ± 32)U/L, (590 ± 70)U/L and (430±52)U/L, respectively;serum levels of AMS were (814±80)U/L, (5031 ± 471) U/L and (2843 ± 236) U/L, respectively, serum levels of IL-6 were (26.7 ± 5.7) pmol/L, (218.0 ±4.7)pmol/L and (112.3 ± 8. O) pmol/L, respectively;serum levels of IL-12 were (4. 2 ± 1.0) pmol/L,(309.5 ± 8.5) pmol/L and (153.7 ± 6.3) pmol/L. The values in ANP group and lipsomal clodronate group were significantly higher than those in control group, while the values in lipsomal clodronate group were significantly lower than those in ANP group (P < 0. 01). Pathologic changes of liver and pancreas were significantly attenuated in lipsomal clodronate group. Conclusions Intravenous liposomal clodronate could exert protective effects on the hepatic injury in rats with ANP.

Objective To investigate the apoptosis of Kupffer cell (KC) induced by lipsomal clodronate in rat with acute necrotizing pancreatitis (ANP). Methods Lipsomal clodronate was prepared by means of thin film, the model of ANP was established by injection of 5% sodium taurocholate of 4 ml/kg into the pancreatic capsule. The Kupffer cells were obtained from ANP rat. After exposure to different doses of lipsomal clodronate (0, 50, 100, 150 μl) , then the proliferation and apoptosis of KC was measured by MTT, flow cytometry and agarose gel electrophoresis of DNA. Results The prepared lipsomal clodronate had an average size of 100～200 nm, the spherical shape of liposome was uniform and confirmed by transmission electron microscope. When exposed to different concentration of lipsomal clodronate for 24 h, the growth suppression rate was 17. 4% , 24. 2% and 31. 1% , respectively, while the apoptosis rate of the KC was (14. 12 ±0.37)% , (18.74±0.43)% and (27.51 ±0.39)%, respectively; the difference was statistically significantly (P<0. 01) , the DNA of KC began degradation and gradually showed clear and characteristic ladder. Conclusions Lipsomal clodronate could induce apoptosis and suppress the growth of Kupffer cells in ANP rats.

Objective To investigate the protective effects of lipsomal clodronate on renal injury in rats with severe acute pancreatifis and the assessment of renal injury. Method Totally 48 rats were randomly divided into three group:normal control group (C);SAP group, in which rats were treated with pure liposomal (P);treatment group, in which SAP rats were treated with liposomal clodronate disodium(T). The SAP model of rat was induced by injection of 5 % sodium taurochohte beneath the pancreatic membrane. Rats of normal control group received isovolumetric injections of 0.9% physiological saline solution instead of sodium taurocholate. Blood samples were collected to measure AMS,BUN,Cr,IL-6 and IL-12 at 2 hors, 6 hours after SAP. At the same time, the samples of pancreatic and renal tissues were taken for observing the pathological changes. Results Compared with controlgroup, serious renal and pancreatic damages were found in group P, and the AMS, BUN, Cr levels elevated signifi-candy (P < 0.01). Compared with group P,the renal and pancreatic damages were attenuated in group T, and the levels of Cr and AMS decreased significantly (P < 0.01), and the IL-6, IL-12 were decreased at 2 hours and 6 hours (P < 0.01). The BUN decreased significantly at6 hours (P < 0.05). Conciusions Excessive release of inflammatory mediator play an important role in renal injury in SAP. Lipsomal clodronate disodium can alleviate the damage of pancreas and kidney.

Objective To investigate the expression of Bcl-2,survivin and pancreatic cancer stem cells markers Oct-4 and ABCG2 in pancreatic cancer cells resistance to chemoradiotherapy.and explore its mechanism.Methods Concurrent ehemoradiotherapy was used to obtain pancreatic cancer cells resistant to chemoradiotherapy,the pancreatic cancer cells without chemoradiotherapy treatment were used as control.Western-blot was applied to detect the expression of Bcl-2,survivin,Oct-4,ABCG2.Results The expression of Bcl-2 was 0.7955±0.0326,0.5718±0.0212,0.6137±0.0382 and 0.8733±0.0461,respectively;the expression of survivin protein was 0.8207±0.0490,0.6973±0.0211,0.7967±0.0346 and 0.8013±0.0398,respectively;the expression of Oct-4 protein was 0.8728±0.0177,0.7861±0.0139,0.4794±0.0932 and 0.4216±0.1043,respectively;the expression of ABCG2 protein was 0.7810±0.1370,0.4957±0.1126,0.6102±0.1358 and 0.4670±0.1274,respectively.in resistant pancreatic cancer cells of SW1990,BxPC3,pc3,jf305 cell line.The corresponding values in the control group were 0.4723±0.018,0.2954±0.0103.0.3587±0.0201 and 0.2718±0.0136;0.4717±0.0274,0.3587±0.0113,0.3891±0.0147 and 0.3326±0.0124;0.6053±0.0142,0.4236±0.0086.0.2385±0.0671 and 0.1985±0.0582;0.3156±0.0582.0.2360±0.0423,0.2813±0.0512 and 0.1808±0.a0370.The expression of all the four proteins significantly increased after ehemoradiotherapy(P＜0.05).Conclusions Pancreatic cancer cells resistant to chemoradiotherapy may contain cancer stem cells.

Objective To investigate the protective effect of clodronate SPIO liposomes on liver injury of rats with severe acute pancreatitis(SAP)and the role of MRI in evaluating the extent of liver injury.Methods Superparamagnetic Fe3O4 nanoparticles were prepared by chemical coprecipitation.Clodronate-SPIO-containing liposomes was prepared by the thin-film method.SAP models were prepared by a uniform injection of sodium taurocholate(2 ml/kg body weight)into the subcapsular space of the pancreas.SD rats were randomly divided into control group,SAP plus SPIO group, and clodronate-SPIO-containing liposome group.Six hours after SAP models were available,T2-weighted MRI scanning(in the same plane)of the liver of rats in each group were performed.At the end of the scanning,blood samples were taken from the supcrior mesenteric vein to measure the contents of serum ALT and AST.Meanwhile, The pathological changes in the liver and pancreas were observed.Results Transmission electron microscopic examination showed that liposomes had a uniform size.No changes in the pancreas of rats in control group were noted.The pathological changes in the pancreas and liver of rats in SAP plus clodronate-SPIO-containing liposome group were significantly milder than those in SAP plus SPIO liposome group.The contents of serum ALT and AST in rats in SAP plus SPIO liposome group were significantly higher than those in control group(P＜0.01), while the contents of serum ALT and AST in rats in SAP plus clodronate-SPIO-containing group were significantly lower than those in SAP plus SPIO liposome group(P＜0.01).The MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus clodronate-SPIO-containing liposome group was significantly lower than that in control group.The significant changes in the MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus Clodronate-SPIO liposome group were noted(P＜0.01).Conclusion Clodronate-containing liposomes have protective effects against liver injury in SAP rats and SPIO can be used as a tracer for MRI examination.

Objective To investigate the effect of triggering receptor expression in myeloid cells-1 (TREM-1) on intestinal macrophage apoptosis in rat.Methods In vitro,the achieved rat intestinal macrophages were divided into 3 groups:control group,LPS (Lipopolysaccharides) group and LPS + LP17 group (n =6 holes of culture plate in each).The concentrations of LPS and LP17 were 1 mg/L and 0.1 mg/L,respectively.The intestinal macrophage apoptosis was measured by using TUNEL kit and flow cytometry after culture for 6 h.All data were statistically analyzed using SPSS 18.0 software.Results The shape and growth of rat intestinal macrophages were quite favorable after culture.The membrane marker of intestinal macrophages,CD14 was clearly observed under immunofluorescence.After macrophage was treated with specific procedure,the cell apoptosis found in LPS group (44.33 ± 7.74)％ was significantly higher than that in control group (19.17 ± 6.01) ％ (P=0.000) measured by TUNEL;the cell apoptosis in LPS +LP17 group (28.33 ± 6.53)％ apparently reduced compared with LPS group (44.33 ±7.74) ％ (P =0.004);there was no significant difference in cell apoptosis between control group (19.17 ± 6.01) ％ and LPS + LP17 group (28.33 ± 6.53) ％ (P =0.050).By flow cytometry,the apoptotic cells in LPS group (16.47 ± 1.66) ％ was significantly increased compared with control group (7.70 ± 1.52) ％ (P =0.000);apoptotic cells in LPS + LP17 group (11.47 ± 3.12) ％ was significantly reduced in comparison with LPS group (16.47 ± 1.66) ％ (P =0.018).There was no significant difference in apoptotic cells between control group (7.70±1.52)％ and LPS + LP17 group (11.47±3.12) ％ (P =0.061).Conclusion LP17 can inhibit TREM-1 expression in intestinal macrophages and reduce intestinal macrophage apoptosis.