OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

OBJECTIVETo observe the anti-inflammatory effect of total saponins of Panax japonicus on LPS-induced RAW264. 7 macrophages.

METHODThe effect of total saponins of P. japonicus of different concentrations on RAW264. 7 cell viability was determined with the MTT method. The NO kit assay was adopted to detect the NO release of total saponins of P. japonicus to LPS-induced RAW264. 7 cells. The enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). The reverse transeriptase-polymerase chain reaction (RT-PCR) was used to determine the expression of inducible nitric oxide synthase (iNOS) ,TNF-alpha,IL-1beta. The protein expression of nuclear transcription factor-kappaB p65 (NF-kappaB p65) was tested by Western blot.

RESULTThe safe medication range of total saponins of P. japonicus was less than 80 mg x L(-1). Compared with the LPS model group, total saponins of P. japonicus high, middle and low dose groups (0.1, 1, 10, 40 mg x L(-1)) could significantly reduce the secretion of NO, TNF-alpha, IL-1beta of LPS-induced RAW264. 7 cells, and inhibit the expressions of iNOS, TNF-alpha and IL-1beta mRNA and the protein expression of NF-kappaB p65.

CONCLUSIONThis study preliminarily proves the protective effect of total saponins of P. japonicus on LPS-induced RAW264.7 macrophages. Its action mechanism may be related to NF-kappaB signal pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; Interleukin-1beta ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Nitric Oxide ; immunology ; Nitric Oxide Synthase Type II ; genetics ; immunology ; Panax ; chemistry ; Protective Agents ; pharmacology ; Saponins ; pharmacology

OBJECTIVETo explore the classification, choice of surgical procedures and the clinical outcome of surgical management for chronic pancreatitis.

METHODS54 patients with chronic pancreatitis undergoing operation in our hospital from 1983 to 2004 were analyzed retrospectively, who were divided into chronic calcifying pancreatitis and chronic obstructive pancreatitis according to the clinical manifestations.

RESULTSThere were 41 men (76%) and 13 women (24%) with a mean age of 54 years. The cause of chronic pancreatitis was alcohol related in 25 cases (46%), cholelithiasis in 21 (39%), and previous episodes of acute pancreatitis in 18 (33%). Clinical manifestations included abdominal pain in 38 cases (70%), obstructive jaundice in 27 cases (50%). There existed a significant difference in some clinical materials between the two groups of chronic calcifying pancreatitis and chronic obstructive pancreatitis, which might mean the different pathologic basis in the two kinds of chronic pancreatitis. A total of 34 patients underwent nine different operations without perioperative deaths. Both the Puestow procedure and the pancreatoduodenectomy was safe and achieved pain relief in a large percentage of patients, which could also improve the exocrine function whereas the endocrine function remained unchanged. Addition of biliary bypass to the Puestow procedure was suitable for the patients with stenosis of common bile duct. Jaundice was the main manifestation in the patients with the inflammatory mass in the head of the pancreas and Whipple's procedure or other resectional procedures should be performed for them. Only drainage of bile duct had a better outcome for the relief of jaundice, but its effect to pancreas need to be further evaluated.

CONCLUSIONThe clinicopathologic characteristics of obstructive chronic pancreatitis was more variable and the surgical management should be also different for individuals.

Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Pancreatectomy ; methods ; Pancreaticoduodenectomy ; Pancreaticojejunostomy ; Pancreatitis ; classification ; pathology ; surgery ; Retrospective Studies

OBJECTIVETo study the characters of chronic pancreatitis complicated by non-calculous obstructive jaundice, and discuss the methods for differentiation and treatment.

METHODTwenty cases selected from January 1985 to December 2004 were analysed in the fields of differentiation and treatment.

RESULTSAll cases didn't present with typical clinical presentations and radiological features. Jaundice was presented as the main complaint. Stricture of the intra-pancreatic common bile duct was the symbolic radiological feature. Pancreatic disseminated inflammation was verified pathologically in these cases. CT, ultrasound, EUS, ERCP, MRCP and antigen-marker of neoplasm failed to offer the data for differentiation. The diagnosis could only be determined by pathological exam. The obstructive jaundice could be solved by biliary-enteric anastomoses successfully.

CONCLUSIONSThe patients with sole complaint of obstructive jaundice account for 15% of all inpatients with chronic pancreatitis. There exists a direct relationship between the jaundice and the pancreatic inflammation. This disorder should be differentiated from total pancreatic carcinoma, but few differentiated material could be offered by preoperative studies. Pathological result derived from the tissue sample obtained within the exploration would be reliable for diagnosis. The bypass between biliary tract and intestine would be a safe and economical treatment method.

Adult ; Aged ; Anastomosis, Roux-en-Y ; Biopsy, Needle ; Cholangiopancreatography, Endoscopic Retrograde ; Choledochostomy ; methods ; Chronic Disease ; Endosonography ; Female ; Humans ; Jaundice, Obstructive ; diagnosis ; etiology ; surgery ; Male ; Middle Aged ; Pancreaticoduodenectomy ; Pancreatitis ; complications ; diagnosis ; surgery ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the clinical outcome of extended retroperitoneal lymphadenectomy as surgical therapy for adenocarcinoma of the head of the pancreas.

METHODSTwenty patients with adenocarcinoma of the head of the pancreas were treated by standard pancreatoduodenectomy (standard group) between 1994 and 1997, and 46 patients with the same disease underwent extended retroperitoneal lymphadenectomy associated with standard pancreatoduodenectomy (radical group) between 1998 and 2002. Clinical and pathological parameters in both groups were reviewed. The postoperative morbidity, mortality, and survival data were compared.

RESULTSThe mean total number of lymph nodes resected was significantly higher in the radical group than in the standard group (P < 0.05). Of the 46 patients in the radical group, 26.09% (12/46) had metastatic adenocarcinoma in the resected retroperitoneal lymph nodes. There was one perioperative death in the standard group and two in the radical group. Postoperative diarrhea and lymphatic leakage were only observed in the radical group. Transfusion requirements and postoperative morbidity did not differ between the two groups. The 1-, 2-, and 3-year survival rates were 63.16%, 31.58%, and 21.05% in the standard group, and 65.91%, 37.71%, and 21.21% in the radical group (P > 0.05). When the subgroups of patients with positive lymph nodes were analyzed, the 1-, 2-, and 3-year survival rates were 41.67%, 16.67%, and 8.33% in the standard group, and 64.52%, 32.26%, and 12.9% in the radical group (P < 0.05). A trend toward a better survival was observed in the first 2 years after operation in the radical group, but with no significant differences 2 years later.

CONCLUSIONThe addition of an extended lymphadenectomy may improve the early survival without increasing the morbidity, but has no significant effect on long-term survival.

Adenocarcinoma ; mortality ; pathology ; surgery ; Adult ; Aged ; Female ; Humans ; Lymph Node Excision ; methods ; Male ; Middle Aged ; Pancreatic Neoplasms ; mortality ; pathology ; surgery ; Pancreaticoduodenectomy ; Postoperative Complications ; Retroperitoneal Space ; Retrospective Studies ; Survival Rate

BACKGROUNDTissue factor (TF) is overexpressed in many malignant tumours and is linked to the pathogenesis and prognosis of such malignancies. In vitro studies have proved that reduced expression of TF has inhibitory effect on the angiogenesis and cell proliferation of the malignant tumour. Therefore, TF suppression has been raised as a possible treatment for malignant tumours. Here we investigated the effect of celecoxib on TF expression induced by tumour necrosis factor alpha (TNFalpha) in PANC-1 cells and a possible molecular mechanism underlying the celecoxib effect.

METHODSVarious doses of celecoxib solution were added to standard cell numbers of PANC-1 cells mixed with equal dose of TNFalpha for 6 hours. The expression of tissue factor was detected quantitatively by Western blot, whilst the activation of nuclear factor kappaB was tested by electromobility shift assay.

RESULTSAs the doses of celecoxib increased, the tissue factor expression was decreased in PANC-1 cells and so was the activation of nuclear factor kappaB.

CONCLUSIONSCelecoxib can downregulate the expression of tissue factor induced by TNFalpha in PANC-1 cells. This antitumour effect of celecoxib can be explained indirectly via its suppressive role in activation of nuclear factor kappaB.

Celecoxib ; Cell Line, Tumor ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Gene Expression Regulation ; drug effects ; Humans ; NF-kappa B ; metabolism ; Pancreatic Neoplasms ; metabolism ; pathology ; Pyrazoles ; pharmacology ; Sulfonamides ; pharmacology ; Thromboplastin ; genetics ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

BACKGROUNDNowadays, there is a remarkable rise in resectability rate of periampullary adenocarcinoma and the mortality and morbidity of the pancreaticoduodenectomy procedure have been reduced remarkably, while the 5 year survival rates of patients with carcinoma of the head of the pancreas are still below 25%. We conducted this retrospective study to evaluate the clinical outcome of radical pancreaticoduodenectomy with extended retroperitoneal lymphadenectomy as a surgical therapy for adenocarcinoma of the head of the pancreas.

METHODSTwenty cases with adenocarcinoma of the head of the pancreas were treated by standard pancreaticoduodenectomy (removing only the peripancreatic lymph nodes en bloc with the tumour) from 1994 to 1997, and 46 cases with the same disease underwent extended retroperitoneal lymphadenectomy associated with standard pancreaticoduodenectomy from 1998 to 2002. The patients for whom there were insufficient follow-up data, or who had received postoperative adjuvant therapy, were excluded from the analysis. Clinical and pathological parameters of both groups were reviewed. The postoperative morbidity, mortality and survival data were compared statistically.

RESULTSDemographic and histopathological characteristics were similar in the two groups of patients. Performance of the extended lymphadenectomy lengthened the procedure. The mean total number of lymph nodes resected was significantly higher in the radical group (P < 0.05). Of the 46 cases in the radical group, 26% (12/46) had metastatic adenocarcinoma in the resected retroperitoneal lymph nodes. There was one perioperative death in the standard group, and two in the radical group. Postoperative diarrhoea and lymphatic leakage were only observed in the radical group. Transfusion requirements and postoperative morbidity rates did not differ between the two groups. The 1-, 2- and 3-year survival rates were 63%, 32% and 21% respectively in the standard group, and 66%, 38% and 21% in the radical group. No statistically significant difference was found between the groups. When subgroups of node positive patients were analysed, the 1-, 2- and 3-year survival rates were 42%, 17% and 8% respectively in the standard group, and 65%, 32% and 16% in the radical group. Better survival was observed in the first 2 years after operation in the radical group, but no survival differences were seen after 2 years post operation.

CONCLUSIONSThe addition of an extended lymphadenectomy to a pancreaticoduodenectomy did not significantly increase morbidity rates, but was associated with an early survival advantage.

Adenocarcinoma ; mortality ; surgery ; Adult ; Aged ; Female ; Humans ; Lymph Node Excision ; Male ; Middle Aged ; Pancreatic Neoplasms ; mortality ; surgery ; Pancreaticoduodenectomy ; Retroperitoneal Space ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo explore the changes in the expressions of glial fibrillary acidic protein (GFAP) and growth- associated protein-43 (GAP-43) in retinal ganglial cells after neural transplantation.

METHODSThirty-nine rats were randomized into normal control group, nerve amputation group and nerve amputation with peripheral nerve transplantation group. Immunohistochemistry was used to detect the changes in the expressions of GFAP and GAP-43 at different time points after the operations, and real-time PCR was employed to detect the mRNA expressions of 13 genes in the retinal ganglial cells of the rats.

RESULTSImmunohistochemistry showed obviously increased GFAP expressions in the retina following the nerve amputation. GFAP expression was down-regulated while GAP-43 expression upregulated in the retinal ganglial cells after peripheral nerve transplantation. Real-time PCR results showed that 5 days after the operations, retinal GFAP and GAP-43 expressions increased significantly in the nerve amputation group and peripheral nerve transplantation groups as compared with those in the control group, but GAP-43 expression decreased significantly in the former two groups afterwards.

CONCLUSIONThe regenerated retina may adjust the production of GFAP. The retinal ganglial cells express GAP-43 during retinal regeneration. Up-regulation of the expression of GAP-43 provides the evidence for nerve regeneration following the nerve transplantation.

Animals ; Axons ; Female ; GAP-43 Protein ; genetics ; metabolism ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Nerve Regeneration ; genetics ; Optic Nerve ; transplantation ; Optic Nerve Injuries ; metabolism ; Random Allocation ; Rats ; Retinal Ganglion Cells ; metabolism

OBJECTIVETo explore the effect of kallikrein-binding protein (KPB) in protecting retinal ganglion cells (RGCs) and promoting axonal regeneration following optical nerve injury in rats.

METHODSCrush injury of the optic nerve at 0.5-1.0 mm from the eyeball was induced in rats, which received subsequent KBP injection into the vitreous cavity (experimental group) and PBS injection (control group). At 7, 14 and 21 days after the injury, the rats were sacrificed and frozen sections of the eyeball were prepared to observe the structure and thickness of the retina and count the number of survival RGCs with HE staining. The optic nerves were collected for Western blotting to assess the effect of KBP on the RGCs and axonal regeneration.

RESULTSRGC counts and retinal thickness showed significant differences between the two groups. Western blotting also demonstrated a significant difference in the expression of the nerve regeneration marker protein GAP-43 between the two groups.

CONCLUSIONKBP offers protection on RGCs and promotes regeneration of the optic nerve axons after optic nerve injury in rats.

Animals ; Axons ; physiology ; Female ; GAP-43 Protein ; metabolism ; Nerve Regeneration ; drug effects ; physiology ; Neuroprotective Agents ; pharmacology ; Optic Nerve Injuries ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; drug effects ; physiology ; Serpins ; pharmacology

Flavonol synthase (FLS) is one of the key enzymes in flavonoids metabolic pathways. In this study, middle sequence was obtained from Carthamus tinctorius transcriptome sequencing results. Full-length cDNAs of FLS was cloned from petals of C. tinctorius to FLS by using RT-PCR and RACE technology. Its full-length cDNA was 1,201 bp, with an open reading frame of 1,101 bp and 336 encoded amino acids. The phylogenetic analysis showed that, FLS gene encoded amino acids in C. tinctorius were highly homologous with amino acids in congeneric Compositae species, especially Rudbeckia laciniata. The pBASTA-FLS plant expression vector was successfully built by the molecular biology method, which lays a foundation for further studying biology functions of the gene and biosynthesis mechanism of flavonoids.
Amino Acid Sequence ; Base Sequence ; Carthamus tinctorius ; classification ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; genetics ; metabolism