Objective To evaluate the influence of SP600125,a selective c-Jun N-terminal kinase (JNK) inhibitor,on the levels of serum total bile salt (TBA) and Bsep,Ntcp expression in the hepatic tissue of rats with ethinylestradiol induced intrahepatic cholestasis.Methods Rats pregnant for 15days were administered the subcutaneous injection of 17- b -estradiol propylene ( EE ) to modulate the ICP animal models,and be SP600125 to intervene.Testing the level of serum TBA and the expression of c-Jun,Bsep,Ntcp in the hepalatic tissue.Results The average gray values of c-Jun in the group of ICP models were significantly lower than the normal control group ( 101.05 ± 5.20 vs 118.99 ± 5.95,P ＜ 0.05 ).After the intervention of SP600125,comparing with the group of ICP models,the expression of Bsep,Ntcp in the group of SP600125 intervention were significantly higher,and this change in the high dose of SP600125 intervention group was more obvious ( low dose intervention group Bsep:0.452 ±0.031 vs 0.291 ±0.043,Ntcp:0.462 ± 0.015 vs 0.285 ± 0.021,P ＜ 0.05 ; high dose intervention group Bsep:0.568 ± 0.038 vs 0.291 ±0.043,Ntcp:0.605±0.020 vs 0.285 ±0.021,P ＜0.05),while the level of TBA in the serum was significantly lower.Conclusions Treatment with SP600125 can down-regulate the level of c-Jun/AP-1,and it may participate in the lower expression of Bsep、Ntcp in the ICP rats which were induced by 17-bestradiol.

The anti-cancer effects of betulinic acid (BA) on Jurkat cells and its in vitro mechanism were examined by using MTT assay. Apoptosis was detected by using Hoechst33258 staining and annexin-V/PI double-labeled cytometry. The effects of betulinic acid on the cell cycle of Jurkat cells were studied by propidium iodide method. RT-PCR and Western blotting were used to analyze the changes of cyclin D3, bcl-xl mRNA and protein levels in Jurkat cells after treatment with betulinic acid. Our results showed the proliferation of Jurkat cells was decreased in betulinic acid-treated group with a 24-h IC50 value being 70.00 mumol/L. Betulinic acid induced apoptosis of Jurkat cells in a time- and dose-dependent manner. The number of Jurkat cells treated with betulinic acid showed an increase in G(0)/G(1) phase and decrease in S phase. After treatment with 0, 20, 60, 100 mumol/L betulinic acid for 24 h, the number of Jurkat cells was increased from (31.00+/-1.25)% to (58.84+/-0.32)% in G(0)/G(1) phase, whereas it was decreased from (61.45+/-1.04)% to (35.82+/-1.95)% in S phase. PBMCs were less sensitive to the cytotoxicity of betulinic acid than Jurkat cells. The expressions of cyclin D3, bcl-xl mRNA and protein were decreased sharply in Jurkat cells treated with betulinic acid. It is concluded that betulinic acid is able to inhibit the proliferation of Jurkat cells by regulating the cell cycle, arrest cells at G(0)/G(1) phase and induce the cell apoptosis. The anti-tumor effects of betulinic acid are related to the down-regulated expression of cyclin D3 and bcl-xl.
Antineoplastic Agents, Phytogenic/*pharmacology ; Apoptosis/*drug effects ; Cell Proliferation/*drug effects ; Cyclin D3/metabolism ; Down-Regulation/drug effects ; Jurkat Cells ; Triterpenes/*pharmacology ; bcl-X Protein/metabolism

In order to investigate the anti-cancer effects of deguelin and on K562 and K562/ADM cells in vitro and the underlying molecular mechanism and compare the cytotoxicity of deguelin on K562, K562/ADM cells and human peripheral blood mononuclear cells (PBMCs). The effects of deguelin on cell proliferation were assessed by MTT assay. Apoptosis were detected by Annexin V/PI double-labeled cytometry. The effects of deguelin on the cell cycle were studied by a propidium iodide method. Our study showed that deguelin inhibited the proliferation of K562 cell and K562/ADM cell in a time- and dose-dependent manner and had minimal effects on normal human peripheral blood mononuclear cells. The ratio of IC(50) value of deguelin of 24 h on K562/ADM cells to K562 cells was only 1.27, which was significantly lower than the ratio of IC(50) value of ADM (higher than 20). Deguelin could induce apoptosis of K562 cells and K562/ADM cells. K562 cells were arrested at G(2)/M phase while K562/ADM cells were arrested at G(0)/G(1) phase. Our results suggested that deguelin was a novel anti-leukemia agents with high efficacy and low toxicity and it is also a promising agent for reversing drug resistance.

Objective To investigate the influence of acpuncture on free calcium in rat brain cells after focal cerebral ischemia reperfusion.Methods 145 adult male SD rats were randomly divided into control group,simple ischemia reperfusion group and acupuncture with ischemia reperfusion group.The middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established by the modified Longa occlusion method. ①The part of free calcium in rat brain cells,focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 15 min.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 3h, 6h and 12h, the rat was killed and its brain cells were made into single cell suspension,marked by Fluo-3/AM.The fluorescence optical density was recorded by laser confocal microscopy.②The part of nerve functional reconstruction, focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 12 hours.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 7 d, 14 d,30 d,60 d and 90 d, the rat was forced to detect it's strength of the dog.Results ①Free calcium in rats of acupuncture therapy group(6h:10.96±1.18;2h:20.9±4.37) was significantly less than that in control group in 6 h and 12 h after reperfusion (6 h: 16.87 ± 3.56,12 h: 34.10 ±1.06)(P<0.05).②The dog in rats of acupuncture therapy group was significantly more than that in control group in 7 d, 14 d after reperfusion (P＜ 0.05 ).No difference of the dog was detected in 30 d ,60 d and 90 d after reperfusion between the two groups.Conclusion Acupunture could decreases the concentration of free calcium and the expression of Caspase-3 mRNA in rat brain cells after focal cerebral ischemia reperfusion, and it can facilitate the recovery of nerve function.

Objective To investigate the change of indications of emergency obstetric hysterectomy and the clinical application of intraoperative interventions. And to provide evidence for prevention of hysterectomy and improvement of obstetric quality. Methods Clinical data were collected from 97 patients who received emergency obstetric hysterectomy at Shengjing Hospital of China Medical University between January 1st, 2004 and December 31st, 2013. The patients were divided into two groups by the time point of January 1st, 2009: the first group was cases treated between January 1st, 2004 and December 31st, 2008, while the second group was cases treated between January 1st, 2009 and December 31st, 2013. The clinical indicators, surgical indications, intraoperative interventions, and blood loss between the two groups were analyzed retrospectively. Results (1) Incidence:54 857 women delivered at Shengjing Hospital of China Medical University between January 1st, 2004 and December 31st, 2013. Of them, 97 patients received emergency obstetric hysterectomy, with an incidence of 0.177% (97/54 857). (2) The 17 patients delivered vaginally (18%,17/97) and 80 by caesarean section (83%,80/97). Forty-nine patients experienced repregnancy with scar uterus (51%, 49/97). About 41 patients underwent abdominal total hysterectomy (42%,41/97) and 56 received subtotal hysterectomy (58%,56/97). (3) The number of patients were comparable between the two groups (50 vs 47;P>0.05). (4) The main surgical indication was uterine inertia (45%, 44/97). The main causes of uterine inertia were excessive uterine tension (45%, 20/44) and placental abruption due to gestational hypertension (32%, 14/44). Of all the indications, 29 patients in the first group (58%, 29/50) and 15 patients in the second group (32%, 15/47) suffered from postpartum hemorrhage. Pathological placenta embedment occurred in 15 patients in the first group (30%, 15/50) and 25 patients in the second group (53%, 25/47). The incidences of postpartum hemorrhage due to uterine inertia or pathological placenta embedment were significantly different between the two groups (both P<0.05), respectively. (5) In the first group, the average preoperative blood loss was (2 900±1 900) ml, and the average intraoperative amount of infused white&red blood cells was (5.9±3.5) U, with the average operation time of (2.2 ± 1.8) hours and the average in-hospital duration of (7.8 ± 2.3) days. In the second group, the average preoperative blood loss was (3 100± 2 200) ml, and the intraoperative amount of infused white&red blood cells was (6.2± 5.2) U, with the average operation time of (2.5± 2.1) hours and the average in-hospital duration of (7.9 ± 2.9) days. There was no significant difference between the two groups in any of these indicators (P>0.05). Postpartum hemorrhage was usually treated with uterine packing in the first group, but was preferentially treated with potent uterine contraction agents, arterial ligation, uterine balloon compression or B-Lynch suture in the second group. The therapeutic effects of these new treatments were significantly better than uterine packing (P<0.05). Conclusions The incidence of emergency obstetric hysterectomy did not change significantly in the past decade. However, the indications and intraoperative interventions have changed significantly in the second five years compared with the first five years. The main surgical indications were uterine inertia and postpartum hemorrhage due to pathological placenta embedment. Therefore, strict control of caesarean section indications was important to reduce emergency obstetric hysterectomy.

Objective Pancreatic duodenal homeobox-1 ( PDX-1) plays an important role in the occurrence and development of diabetes.More importantly,its potential application in insulin resistance (IR) improvement is attracting further attention .Berberine has been shown to improve glucose metabolism and insulin resistance .In this study,rat models of type 2 diabetes mellitus ( T2DM) were established by combination of intraperitoneal injection of low -dose streptozotocin and highg-lucose-high-fat diet induction .The effect of berberine on fasting glucose , fasting serum insulin , homeostasis model assessment of insulin resistance ( HOMAI-R) , and ex-pression of PDX -1 of islet in T2DM rats were analyzed .The present study aimed to evaluate the anti-diabetic efficacy of berberine and study the mechanism of its preliminary therapeutic effects .Methods Twenty healthy adult male Sprague Dawley ( SD) rats were di-vided by random number table into the normal control group ( n =6) feeding with a standard diet ,and the experimental group ( n =14 ) given an intraperitoneal injection of streptozotocin .The rats with fasting blood glucose ( FBG) greater than 13.8 mmol/L were ran-domly divided into diabetes mellitus group feeding high sugar high fat diet for two weeks and then continuing with a standard diet , and the berberine group given berberine [180 mg /(kg· d)] every day.The normal control group and diabetes mellitus group were given physiological saline for every day 6 weeks.At the end of 6th week, the rats were executed , and the levels of fasting glucose, fasting se-rum insulin, HOMA-IR, and expression of PDX-1 in pancreas islet of each group were detected .The expression of PDX1-in pancreas islet was examined by immunohistochemistry .The image pro plus 6.0software was used to calculate the integrated optical density ( IOD) of positive expression .Results Compared with the normal control group , FBG, fasting insulin levels ( FINS) , and HOMA-IR of the diabetes mellitus group and berberine group were significantly increased ( P <0.05 ) .Compared with the diabetes mellitus group, FBG, FINS, and HOMA-IR of the berberine group were statistically significantly decreased [ ( 13.11 ±6.87 ) mmol/L vs (18.45 ±4.69) mmol L/, (2.58 ±0.34) μIU/ml vs (2.98 ±0.40) μIU/ml, 1.60 ±0.91 vs 2.75 ±0.28, P <0.05].Compared with the normal control group , the expression of PDX-1 of the islet in the diabetes mellitus group and berberine group was statistically significantly decreased ( P <0.05).After treatment with berberine, FBG, FINS, HOMA-IR, and IOD of PDX-1 in pancreas islet of berberine treatment rats were significantly increased compared with the diabetes mellitus group ( 9.14 ±1.51 vs 6.79 ±0.98 , P <0.01 ) .Conclusions The increases of PDX-1 in islet may be one of the mechanism in the improvement of hyperglycemia .

Objective Estimate the paternity exclusion probability for 15 short tandem repeats (CODIS set plus Penta D and Penta E) in duo paternity tests. Methods 644 random individuals were paired to con-struct 207046 fictitious duo families. Cases of non-exclusion or with only one exclusionarylocus were count-ed. Results 27 cases could not be excluded by the 15 STR loci. The combined probability of exclusion was 0.999869. Number of cases showing only one exclusionary locus was 384 (0.185%). Conclusion To con-firm relationship in duo cases with only 15 STRs should be careful. Extensive genetic markers is needed to discriminate one locus exclusion or mutation.

Objective To study the characteristic of chromogranin A(CgA),synaptophysin(SYN) and neuron-specific enolase(NSE) in cortex and medulla of adrenal.Methods Immunohistochemical method was used to detect the expression of CgA,SYN and NSE in routinely processed tissue specimens from human adrenal neoplasm and hyperplasia.Results The expression of CgA was detected in cortex of adrenocortical adenoma,adrenal neoplasm and chromophile tumor,but not in that of hyperplasia and normal adrenal.The expression of NSE was detected in cortex of adrenal cortical hyperplasia,cortical adenomas and pheochromocyte carcinoma,but not in that of pheochromocytoma and normal adrenal.The expression of SYN was detected in cortex of corticohyperplassia,cortical adenomas,medull neoplasm,chromophile tumor and pheochromocyte carcinoma,but not in that of cortical adenocarcinoma and normal adrenal.The expressions of CgA,NSE and SYN were detected in the medulla of all cases.Conclusion The characteristic of CgA,SYN and NSE in cortex and medulla of adrenal are useful in the differential diagnosis of tumor and hyperplasia in the adrenal.The relationship between the adrenal cortex and medulla is close.Besides the classical hypothalamus-pituitary-adrenal axis(HPAA) and rennin-angiotensin-aldosterone system(RAAS),the adrenal cortex and medulla have the basis of anatomy and endocrinology.

Objective To study the effect of SDF1/CXCR4 and FN/?1 integrin modulated by curcumin on the adhesion and migration of B lymphoma Raji cell and to explore the mechanism of curcumin to inhibit the invasiveness of lymphoma.Methods The experiment consisted of eight groups: Control,FN,SDF,FN+anti-?1 integrin,SDF+anti-CXCR4,curcumin+FN,curcumin+SDF.Adhesion assay and migration assay were used to detect the difference of invasiveness among different groups.Results In vitro,SDF1 and FN can increase the adhesion and migration of Raji cells,which can be blocked by ?1 integrin antibody and CXCR4 antibody.Curcumin can inhibit the adhesion and migration of Raji to FN in a dose-dependant manner.Low dosage curcumin had minimal effect on the adhesion and migration of Raji induced by SDF except that 25?mol/L curcumin can inhibit the adhesion and migration of Raji.Conclusion FN/?1 integrin and SDF1/CXCR4 pathway play important roles in the adhesion and migration of Raji cells.Curcumin can decrease the invasiveness of Raji cells through inhibiting FN/?1 integrin.It has good prospect of clinical application.

AIM:To investigate the inhibitory effects of triptolide on cell proliferation and metastasis in Burkitt's lymphoma cell line Raji cells.METHODS: The effects of triptolide on the growth of Raji cells were studied by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium(MTT) assay.The effects of triptolide on the cell apoptosis of Raji cells were detected by using Annexin Ⅴ/PI double-labled cytometry.The effects of triptolide on CXCR4 expression on Raji cells were studied by flow cytometric analysis.Chemotaxis assays were performed to observe the effects of triptolide on migration of Raji cells towards recombinant human SDF-1?(rhSDF-1?)in vitro.RESULTS: Triptolide inhibited the proliferation of Raji cells in a dose-and time-dependent way with a 24 h IC50 value of 43.06 nmol/L and a 36 h IC50 value of 25.08 nmol/L.Following the treatment of triptolide,the cell apoptosis rate was increased as the treatment concentration increased and the culture time extended.The effects were dose-and time-dependent.Triptolide could downregulate the expression of CXCR4 on Raji cells in a dose-dependent manner.Moreover,chemotaxis assay suggested that triptolide could block the migration of Raji cells to rhSDF-1? in vitro,and the inhibition was dose-dependent.CONCLUSION: Triptolide could inhibit the cell proliferation and induce the cell apoptosis of Raji cells.Furthermore,it could block the cell metastasis of Raji cells in vitro and the underlying mechanism might be related to the inhibition of the SDF-1/CXCR4 axis.