OBJECTIVE:To synthesize the aryl-pyridazinone acid and curcumin ester. METHODS:With the raw material of 5-methyl-2-(3-chloro-4-fluorophenyl)-2-oxo-pyridazine acid (compound 1) and curcumin,there was ester-forming in the two sides of a phenolic hydroxyl and pyridazin-6-one carboxyl in curcumin by the catalysis of N,N-dicyclohexyl carbodiimide(DCC)/4- dimethylaminopyridine (DMAP). The targeted product,MS and NMR characterization of the structure were obtained through column chromatographic separation. Single factor was adopted to detect the effect of mixture ratio of curcumin and compound 1,re-action temperature and time and catalyst on the reactions. RESULTS:The productivity of targeted product (aryl-pyridazinone acid and curcumin ester)was 56.3%(in curcumin),the content by HPLC was 98.1%. The optimum conditions were as follows as the mixture ratio of curcumin and compound 1 was 1∶3,reaction temperature was 50 ℃,reaction time was 10 h and the catalyst was DCC/DMAP. CONCLUSIONS:The aryl-pyridazinone acid and curcumin ester is successfully synthesized with stable process.

Acute Disease ; Humans ; Lumbar Vertebrae ; Male ; Middle Aged ; Osteochondroma ; diagnosis ; pathology ; therapy ; Spinal Neoplasms ; diagnosis ; pathology ; therapy

Abdominal Injuries ; complications ; Adenosarcoma ; etiology ; Adult ; Cicatrix ; complications ; Endometriosis ; complications ; Female ; Humans

This study was purposed to explore the caspase-independent apoptosis pathway in human multiple myeloma cell RPMI8226 induced by arsenic trioxide (As(2)O(3)). MTT method was used to analyze the proliferation inhibition rate; flow cytometry was used to detect the apoptosis rate; Western blot was used to determine the expressions of BCL-2 and Caspase-3 in RPMI8226 cells. The results showed that As(2)O(3) (0.1 - 20 µmol/L) significantly inhibited the proliferation of RPMI8226 (P < 0.05) in concentration- and time-dependent manner. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, the apoptosis rate of zVAD-fmk (20 µmol/L) and As(2)O(3) combined treated group did not change. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, zVAD-fmk (20 µmol/L) combined with As(2)O(3) (10 µmol/L) treatment group showed significant increase of expressions of Caspase-3 and BCL-2. It is concluded that As(2)O(3) can inhibit the proliferation of RPMI8226 cells. As(2)O(3) can induce apoptosis of RPMI8226 cells, and a caspase-independent process probably exist in As2O3-inducing RPMI8266 cells apoptosis.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

AIM:To investigate the role of peroxisome proliferator-activated receptors(PPARs)-inflammation signaling pathways in diabetic hepatopathy.METHODS:Diabetic mouse model was established by feeding the mice with a high-energy diet for 4 weeks combined with intraperitoneal injection of streptozotocin(STZ;40 mg· kg-1· d-1for 5 d). The hepatopathy model was confirmed by histopathological observation and the indexes of liver function, such as alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP),after another 4 weeks.Mo-reover,fasting blood glucose(FBG), and serum levels of total cholesterol(TC), triglyceride(TG)and insulin were measured,and the HOMA insulin resistance index(HOMA-IR)was calculated.The mRNA and protein expression levels of PPARs and inflammation-related factors were measured by qPCR and Western blot, respectively.RESULTS: After treatment with STZ for 7 d,the FBG of mice exceeded 11.1 mmol/L,suggesting that the diabetic model was established. After 4 weeks,the structural deformation of the hepatocytes(including hepatocytes containing abundant fat vacuoles, and inflammatory cell infiltration),and the increases in the serum levels of insulin,HOMA-IR,TC,TG,ALT,AST and ALP were observed(P<0.01), indicating the occurrence and progression of hepatopathy in diabetic mice.Meanwhile, com-pared with the control group,the mRNA and protein expression of PPARα,PPARβand PPARγdecreased,but the expres-sion of nuclear factor-κB(NF-κB),cyclooxygenase 2(COX-2)and inducible nitric oxide synthase(iNOS)significantly increased in the diabetic hepatopathy mice(P <0.01).CONCLUSION: Down-regulation of PPARα, PPARβand PPARγand activation of NF-κB-COX-2/iNOS signaling pathways may be involved in the diabetic hepatopathy in mice in-duced by long-term high-energy diet feeding combined with intraperitoneal injection of STZ.

Objective To compare the acute toxicities between two prospective, non-randomize phase Ⅱ trials on adjuvant radiochemotherapy of capecitabine with or without oxaliplatin in patients with stage Ⅱ and Ⅲ rectal cancer. Methods From March 2005 to November 2007,based on two fulfilled phase Ⅰ studies,two phase Ⅱ trials were launched respectively to further observe the tolerance and toxicity. In one tria1,118 patients were treated with concurrent capecitabine and radiotherapy (Cap-CRT trial), with radio-therapy of DT50 Gy/25 F/5 wks to the pelvis, and capecitabine at a dose of 1600 mg/m2/d(d1-d14,3 weeks per cycle). In the other trial, 90 patients received concurrent oxaliplatin, capecitabine and radiothera-py(Cap-Oxa-CRT trial), with the same radiotherapy schedule, while oxaliplatin at a dose of 70 mg/m2(d1, d8) and capecitabine of 1300 mg/m2/d(d1-d14,3 weeks per cycle). Results There was no significant difference in the delay of radiotherapy (10.2% vs 6.7%, X2=0.80, P=0.460) or chemotherapy (9.3% vs 19.1%, X2=4.80,P=0.090) between Cap-CRT and Cap-Oxa-CRT trials. Grade 1-4 leukopenia,diar-rhea and nausea were the most common acute side-effects in the both trials, accounting for 70.2%, 65.9% and 42.3%, respectively. When comparing with Cap-CRT trial, Cap-Oxa-CRT trial had significantly more grade 1-4 non-hemotological toxicities, mainly in Gl,including nausea (68.9% vs 22.0%, X2=46.90, P= 0.000), diarrbea(76.7% vs 57.6%, X2=13.50, P=0.009), fatigne(47.8% vs 13.7%, X2=18.90,P= 0.000), hand-foot syndrome (14.4% vs 4.2%, X2=7.10, P=0.029), and inappetence (50.0% vs. 27.9%, X2 = 25.70, P=0.000), but not in hematological toxities of leukopenia, anemia or thrombocytope-nia. Of all the patients,grade 3 and grade 4 toxicities were diarrhea(24.0% and 1.0%),leukopenia(4.3% and 0.0%),radiation-induced dermatitis(3.8% and 0.0%),cramping abdominal pain(1.0% and 0.0%) and fatigue(0.5% and 0.0%). Only grade 3 and 4 diarrhea was significantly more in Cap-Oxa-CRT trial than in Cap-CBT trial(33.0% vs 18.6%, X2=5.90,P=0.023). Conclusions For patients with stage Ⅱ and Ⅲ rectal cancer,both the postoperative concurrent radiochemotherapy regimens are tolerable,though Cap-Oxa-CRT trial has more grade 3 and 4 diarrhea.

In this paper, we studied the relationship between the prostaglandin F(2alpha) (PGF(2alpha))-induced cardiac hypertrophy and calcineurin (CaN) signal transduction pathway in vivo and in vitro. Male Sprague-Dawley rats were given a single i.p. injection with monocrotaline (MCT) (60 mg/kg) and then given orally with celecoxib (20 mg/kg) or vehicle once a day for 14 d before (from d 1 to d 14) or after (from d 15 to d 28) right ventricular hypertrophy (RVH) was formed. Body weight (BW), right ventricular weight (RV), left ventricular with septum weight (LV), as well as lung weight were determined. RVH index (RVHI=RV/LV), RV/BW, and lung weight/BW were calculated and histological changes were observed with transmission electron microscope. PGF(2alpha) level, atrial natriuretic peptide (ANP) and CaN mRNA expressions, expression of CaN and its downstream effectors, NFAT(3) and GATA(4) protein were assayed by EIA kit, RT-PCR, and Western blotting, respectively. The cardiomyocyte hypertrophy in primary culture induced by PGF(2alpha) (0.1 micromol/L) was evaluated by measuring the cell diameter, protein content, and ANP mRNA as well as CaN mRNA expressions. It was found that 14 d or 28 d after MCT was given, the RVHI, RV/BW, and lung weight/BW were significantly increased by 47%, 53% and 118%, and by 64%, 94% and 156%, respectively; at the same time PGF(2alpha) levels in RV tissue were increased by 44% and by 51% with increasing RVHI, and elevated expressions of ANP and CaN mRNA, as well as CaN, NFAT(3) and GATA(4) proteins in a positive correlation manner. Furthermore, some histological injuries were found in RV tissue. Celecoxib, a cyclooxygenase inhibitor, obviously blunted the elevation of RVHI, RV/BW, and lung weight/BW no matter it was given before or after RVH. In vitro experiments showed that 0.1 micromol/L PGF(2alpha) significantly increased the cardiomyocyte diameter and protein content, and promoted ANP and CaN mRNA expressions, which was blocked by cyclosporin A, a CaN inhibitor. Our results indicate that PGF(2alpha) may be involved in cardiac hypertrophy induced by MCT in rats through CaN signal transduction pathway.
Animals ; Calcineurin ; genetics ; metabolism ; physiology ; Cells, Cultured ; Dinoprost ; metabolism ; physiology ; Hypertrophy, Right Ventricular ; chemically induced ; metabolism ; physiopathology ; Male ; Monocrotaline ; Myocytes, Cardiac ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

OBJECTIVETo study the influence on expression of interstitial collagen in lung of rats exposed to ammonium perchlorate.

METHODSThe rats were treated with AP by intratracheal instillation and sacrificed after 3 d, 7 d, 14 d, 28 d. The mRNA level of collagen I and collagen III in the lung tissues was measured by RT-PCR.

RESULTSThe levels of collagen I on 7 d, 14 d, 28 d exposed for high dose group (1.93 +/- 0.41, 3.50 +/- 0.90, 2.33 +/- 1.12) and 28 d exposed for medial dose group (2.58 +/- 0.86) were higher significant (P < 0.05) than those in negative control group (0.52 +/- 0.11, 0.77 +/- 0.15, 0.86 +/- 0.29) The levels of collagen I in low dose group exposed for 14 d(1.99 +/- 0.67), 28 d(1.85 +/- 0.67) and high dose group 14 d(3.50 +/- 0.90) exposed for 14 d were higher significant (P < 0.05) compared to those exposed to AP for 3 d(0.52 +/- 0.14), (1.71 +/- 0.38). The levels of collagen III on 14 d, 28 d exposed for high dose group (2.60 +/- 1.00, 1.46 +/- 0.36) and 14 d, 28 d exposed for medial dose group (1.80 +/- 0.51, 2.16 +/- 0.87) were higher significant (P < 0.05 or P < 0.01) than those in negative control group(0.54 +/- 0.20, 0.52 +/- 0.22); The levels of collagen III in medial dose group(2.16 +/- 0.87) exposed for 28 d, and high dose group exposed for 14 d (2.60 +/- 1.00) were higher significant (P < 0.05) compared to those exposed to AP for 3 d(1.22 +/- 0.32, 0.96 +/- 0.17).

CONCLUSIONThe results suggest that AP has a toxic effect to promote the expressions of collagen I and collagen III mRNA in lungs of rats, and may be cause fibrosis, but there should have more suffice evidences to prove that AP is exactly compound that made lung fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Female ; Lung ; drug effects ; metabolism ; pathology ; Perchlorates ; toxicity ; Pulmonary Fibrosis ; metabolism ; Quaternary Ammonium Compounds ; toxicity ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of endothelial progenitor cell (EPC) injection in the restoration of vascular niche in bone marrow (BM) after allo-BMT in mice, and to observe its role on hematopoietic reconstitution.

METHODS6-8 weeks old female BALB/c (H-2(d)) were randomized to BMT (allo-BMT) group and combined EPC transplant (allo-BMT + EPC) group. For allo-BMT group, female BALB/c mice were lethally irradiated with 60Co source, and then were injected intravenously with 5×10(6) BM cells from donor mice. In allo-BMT + EPC group, recipient mice were injected intravenously with 5×10(6) BM cells and 5×10(5) EPC from donor mice. The recipients were monitored for histological changes of endothelial cells (EC) in BM. The recovery of hematopoiesis was determined by white blood cell counts and the proportion of reticulocytes in circulation and the proportion of hematopoietic stem cells (HSC) in BM. The histology of hematopoiesis in BM was also detected.

RESULTSThe in vitro induced EPC successfully homed to the bone marrow of recipients. The ECs of allo-BMT recipients were destructed severely, while the structures of ECs were restored in EPC treated recipients. 10 and 15 days after allo-BMT, the amount of Lin-c-kit(+)Sca-1(+) cells in the BM of the EPC treated group were (20.31 ± 2.65)×10(3) per mouse and (10.26 ± 2.19)×10(3) per mouse, while the allo-BMT group's were (9.61 ± 0.98)×10(3) per mouse and (4.09 ± 1.34)×10(3) per mouse; and 15 days after allo-BMT, the amount of white blood cell counts and proportion of reticulocytes of the EPC treated group were (1.20 ± 0.11)×10(9)/L and (2.35 ± 0.30)% comparing to the allo-BMT group which were (0.65 ± 0.10)×10(9)/L and (1.63 ± 0.20)%.

CONCLUSIONCo-transfer of donor EPC restores the ECs of bone marrow, which consequently promotes hematopoietic reconstitution in murine allo-BMT.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Endothelial Cells ; cytology ; Female ; Leukocyte Count ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Reticulocyte Count ; Stem Cell Niche ; Stem Cells ; cytology

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism