Dictamnine inhibits pancreatic cancer cell growth and epithelial-mesenchymal transition by blocking the PI3K/AKT signaling pathway.

Many different treatments are available for pancreatic cancer (PC), including surgical resection, chemotherapy, radiotherapy, and immunotherapy, but these treatments are often ineffective at curing PC. Hence, identifying new and effective agents or strategies to improve therapeutic effects is critical. This study focused on the efficacy of dictamnine (DTM), a furan quinoline alkaloid extracted from Dictamnus dasycarpus Turcz., in treating PC. Our in vitro results showed that DTM can mitigate cell proliferation and induce cell cycle arrest and apoptosis in two different human PC cell lines. Moreover, epithelial-mesenchymal transition (EMT) was prevented during DTM treatment, reflected by reduced cell migration and invasion abilities. In vivo studies demonstrated that DTM treatment led to a remarkable inhibition of tumor growth in a xenograft nude mouse model. Mechanistic investigation showed that DTM might act by restraining constitutive and induced PI3K/AKT activity. In summary, our results demonstrated that DTM slows PC progression by inhibiting the activity of the PI3K/AKT signaling pathway and its downstream effectors and that DTM is effective as a pathway-specific cancer therapy. These findings could provide a greater understanding of the function of anticancer drugs and new options for PC treatment.

microRNA expression alteration after arsenic trioxide treatment in HepG-2 cells.

BACKGROUND AND AIM: More and more microRNA (miRNA) are found to be involved in tumor genesis and progress. Arsenic trioxide has been an effective chemotherapeutic drug in cancer therapy for many years. In this study, we aimed to find the miRNA involved in the mechanisms of arsenic trioxide treatment in cancer therapy. METHODS: We detected the expression profile of miRNA by miRNA microarray and quantitative real-time polymerase chain reaction. Cell viability assay, flow cytometry analysis, prediction of miRNA targets, Western blot analysis and luciferase reporter assay were carried out to determine the role of one selected miRNA, namely mir-29a, in affecting the biological behaviors of HepG-2 cells. RESULTS: Among the 677 human miRNA in the microarray, five miRNA were upregulated and four were downregulated in HepG-2 cells treated with arsenic trioxide compared to their controls. If only changes above two folds were considered, four miRNA were identified, namely miR-24, miR-29a, miR-30a and miR-210, which were all upregulated. Among them, miR-29a showed a positive therapeutic effect in liver cancer cells by inhibiting cell growth and inducing cell apoptosis, and PPM1D was confirmed to be the target gene of miR-29a. Furthermore, a synergy effect was detected between miR-29a and arsenic trioxide. CONCLUSIONS: Arsenic trioxide altered miRNA expression profile in HepG-2 cells. Among the altered miRNA, miR-29a seemed to take a role in the mechanism of arsenic trioxide in liver cancer therapy. The synergy effect between miR-29a and arsenic trioxide may offer this drug a new chance in cancer therapy by decreasing its dose and toxic side-effects.

Alterations of bone marrow sinusoidal endothelium in rat and patients with liver cirrhosis.

Whether bone marrow changes occur and potentially contribute to the hematological abnormalities in liver cirrhosis remain unclear. In this study, we established a rat model of liver cirrhosis induced by carbon tetrachloride. Electron microscopy examination showed focal lesions in bone marrow sinusoidal endothelium and hematopoietic cells in animals with cirrhosis. With the persistence of liver cirrhosis, injuries of bone marrow sinusoidal endothelium progressed from mild mitochondrial changes to nuclear pycnosis and cell disruption, and the trilineage hematopoietic cells showed apoptosis and necrosis. Immunohistochemistry revealed increased expression of E-selectin, P-selectin and vWF in bone marrow sinusoidal endothelium of the cirrhotic rats, which was consistent with the data from semiquantitative reverse transcriptase-polymerase chain reaction analysis. Autopsy specimens from patients with liver cirrhosis (in the absence of other disease) showed the same findings as detected by immunohistochemistry in animal models. The results provide evidence of the association between liver cirrhosis and bone marrow alterations by demonstrating the bone marrow sinusoidal endothelium lesions in both a rat model and patients. It also indicates that activation or injury of bone marrow sinusoidal endothelium mediated by E-selectin, P-selectin, and vWF might have a role in pathogenesis of bone marrow changes during liver cirrhosis. The lesions of bone marrow sinusoidal endothelium might contribute to the hematological abnormalities in the end stage of liver disease.

Induction of apoptosis by artemisinin relieving the severity of inflammation in caerulein-induced acute pancreatitis.

AIM: To observe the apoptosis and oncosis of pancreatic acinar cells and secondary inflammatory reaction in pancreatic tissue from rats with acute pancreatitis (AP), and the influences of artemisinin on them. METHODS: AP was induced by 4 intraperitoneal injections of caerulein at 1 h intervals. To induce apoptosis, solution of artemisinin (50 mg/kg) was given intraperitoneally 1, 12, 24 and 36 h after the last caerulein injection. Histological examination of impairment of pancreatic tissue and detection of serum amylase were performed to evaluate the severity of acute pancreatitis. Apoptosis and oncosis were detected with acridine orange (AO) and ethylene dibromide (EB) staining. Caspase-3 and myeloperoxidase (MPO) activity were measured by colorimetric assay. Nuclear factor-kappa B (NF-kappaB) activation was detected by flow cytometry. Macrophage inflammatory protein-1alpha (MIP-1alpha) protein was measured by Western blot. Interleukin-1beta (IL-1beta) mRNA was detected by RT-PCR. RESULTS: Addition of artemisinin increased the number of apoptotic cells (11.7% +/- 1.4% vs 6.3% +/- 0.7%, P < 0.05), while reduced the number of oncotic cells (13.0% +/- 2.4% vs 17.5% +/- 2.2%, P < 0.05). The activity of caspase-3 speeded up (1.52 +/- 0.21 vs 1.03 +/- 0.08, P < 0.05), the pancreas pathological impairment was relieved (3.0 +/- 0.5 vs 4.0 +/- 0.5, P < 0.05) and the level of serum amylase decreased (5642 +/- 721 U/dL vs 7821 +/- 653 U/dL, P < 0.05). The activation of NF-kB (29% +/- 4.1% vs 42% +/- 5.8%), MIP-1alpha protein (3.7 +/- 0.5 vs 5.8 +/- 0.7), MPO (0.52 +/- 0.06 U/g vs 0.68 +/- 0.09 U/g), IL-1beta mRNA (1.7 +/- 0.3 vs 2.4 +/- 0.4) in the apoptosis inducing group was obviously decreased (P < 0.05). CONCLUSION: Inducing apoptosis can relieve pathological impairment and inflammatory reaction in AP rats.

Regulating effects of arsenic trioxide on cell death pathways and inflammatory reactions of pancreatic acinar cells in rats.

BACKGROUND: It is accepted that inflammatory cytokines play a key role in the development of acute pancreatitis, so blocking the initiation of inflammatory reactions may alleviate pathological changes of acute pancreatitis. We studied the regulatory effect of arsenic trioxide (As(2)O(3)) on apoptosis and oncosis of pancreatic acinar cells in vitro and in vivo and its therapeutic effect on acute pancreatitis. METHODS: Pancreatic acinar cells were isolated by collagenase digestion method. Apoptosis and oncosis of isolated pancreatic acinar cells were detected with Hoechst 33258 + PI or Annexin V + PI double fluorescent staining. Amylase and lactate dehydrogenase release were measured. Acute pancreatitis was induced in Wistar rats by intraperitoneal injections of caerulein, and apoptosis was detected with terminal dUTP nick-end labeling method. Tumor necorsis factor alpha (TNF-alpha) mRNA, myeloperoxidase, nuclear factor-kappaB and histological grading of pancreatic damage were measured. RESULTS: There was an increased apoptosis but a decreased oncosis of pancreatic acinar cell after the treatment with As(2)O(3). The levels of lactate dehydrogenase and amylase release were markedly decreased in As(2)O(3) treated group. Myeloperoxidase content, TNF-alpha mRNA level, nuclear factor-kappaB activation and pathological score in As(2)O(3) treated group were significantly lower than in the untreated group. CONCLUSIONS: As(2)O(3) can induce apoptosis and reduce oncosis of pancreatic acinar cell, thus resulting in reduced release of endocellular enzyme of acinar cells, reduced inflammatory cell infiltration and decreased the production of inflammatory cytokines, so that the outcome of alleviated pathological changes was finally achieved.

[Effects of Astragalus polysaccharides and Cordyceps sinensis mycelium mixture on the transforming growth factor-beta1 expression in chronic rejection of artery transplantation: experiment with rats].

OBJECTIVE: To explore the effect of Astragalus polysaccharides (APS) and Cordyceps sinensis (CP) mycelium mixture on the chronic rejection of artery transplantation. METHODS: The abdominal arteries of 180 Wistar rats were isolated and transplanted to 180 SD rats whose with their abdominal arteries resected. The 180 recipient SD rats were randomly divided into blank control group (n = 20), APS group (n = 40, receiving gastric perfusion of APS 1 mg/kg bid), APS + low-dose CP group (n = 40, receiving APS and CP mycelium powder 1.25 g/d), APS + medium-dose CP group (n = 40, receiving APS and CP mycelium powder 2.5 g/d), and APS + high-dose CP group (n = 40, receiving APS and CP mycelium powder 5 g/d). Thirty days later the SD rats were killed with the transplanted abdominal arteries taken out to undergo microscopy. The expression of transforming growth factor (TGF)-beta(1) in the transplanted artery was examined by immunohistochemistry. RESULTS: In the control group, overexpression of TGF-beta(1) was observed in the intimal smooth muscle cells, monocytes, arterial endothelial cells, and arteriole, venule, and blood capillary endothelial cells in extima. However, the expression levels of TGF-beta(1) in the APS group and the 3 mixture administration groups were all significantly lower, especially in the medium- and high-dose groups (all P < 0.01). Severe edema of arterial endothelial cells and proliferation of monocytes were observed microscopically in the control group. Under electron microscope, rough endoplasmic reticulum and Golgi body were abundant in the control group. While in the mixture administration groups these changes were either weakened or absent. CONCLUSION: Astragalus polysaccharides and Cordyceps sinensis mycelium polysaccharides mixture can decrease the expression of TGF-beta(1) and inhibit the synthesis of collagen in the transplanted arteries.

[The importance of nonrecurrent laryngeal nerve in thyroid surgery].

OBJECTIVE: To investigate the anatomic variation of nonrecurrent laryngeal nerve (NRLN) and its surgical identification and prevention during thyroidectomy. METHODS: The database of 5 NRLN cases was analyzed to investigate the difference of operative maneuvers and procedures. RESULTS: All 5 NRLN were located in the right side. Two cases were found have vocal cord paralysis and 1 case recovered in 3 cases who have NRLN injures. CONCLUSIONS: Any transverse bond should not be cut between vascular and laryngeal except middle thyroid vein. Recurrent laryngeal nerve (RLN) should be dissected during thyroid excision. Cervical pneumogastric nerve should be systematic dissected to detect whether RNLN is exist, if RLN is not exist in the same side.

Expression of cell cycle/growth regulator genes in human hepatocellular carcinoma and adjacent normal liver tissues.

We investigated the gene expression of the cell cycle/growth regulators in hepatocellular carcinoma (HCC) through the usage of Atlas human cancer array membranes, semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Northern blot. Hybridization of cDNA array membrane was performed with alpha-32P-labeled cDNA probes synthesized from RNA, isolated from HCC and adjacent non-cirrhotic normal liver. RT-PCR of 24 paired specimens and Northern blot of 4 paired specimens were used to confirm the expression patterns of the cell cycle/growth regulator genes identified by Atlas array hybridization. Among 79 genes related to cell cycle/growth regulators, transcription factor DP2 (TFDP-2) and E2F-3 were up-regulated, whereas dual-specificity mitogen-activated protein kinase kinase 1 (MAPKK1) and cell division protein kinase 3 (CDK3) were down-regulated in HCC. RT-PCR of TFDP-2 gave result consistent with Atlas human cancer cDNA array findings. Northern blot analysis of TFDP-2 and E2F-3 of 4 paired specimens all showed up-regulation in HCC compared to normal liver tissues. The results obtained from Atlas microarray provided for the first time a liver cancer-specific expression profile, which identified the gene expressions comprehensively and systematically. The findings may lead to better understanding of the mechanism of onset and progression of HCC. The rapid and high-throughout method of profiling gene expression by cDNA array provides an overview of the key factors that may be involved in HCC. Some genes are reported for the first time in HCC.

Current treatment for liver metastases from colorectal cancer.

The liver is the commonest site of distant metastasis of colorectal cancer and nearly half of the patients with colorectal cancer ultimately develop liver involved during the course of their diseases. Surgery is the only therapy that offers the possibility of cure for patients with hepatic metastatic diseases. Five-year survival rates after resection of all detectable liver metastases can be up to 40 %. Unfortunately, only 25 % of patients with colorectal liver metastases are candidates for liver resection, while the others are not amenable to surgical resection. Regional therapies such as radiofrequency ablation and cryotherapy may be offered to patients with isolated unresectable metastases but no extrahepatic diseases. Hepatic artery catheter chemotherapy and chemoembolization and portal vein embolization are often used for the patients with extensive liver metastases but without extrahepatic diseases, which are not suitable for regional ablation. For the patients with metastatic colorectal cancer beyond the liver, systemic chemotherapy is a more appropriate choice. Immunotherapy is also a good option when other therapies are used in combination to enhance the efficacy. Selective internal radiation therapy is a new radiation method which can be used in patients given other routine therapies without effects.

Gene expression profiles of hepatoma cell line HLE.

AIM: To investigate the global gene expression of cancer related genes in hepatoma cell line HLE using Atlas Human Cancer Array membranes with 588 well-characterized human genes related with cancer and tumor biology. METHODS: Hybridization of cDNA blotting membrane was performed with (32)P-labeled cDNA probes synthesized from RNA isolated from Human hepatoma cell line HLE and non-cirrhotic normal liver which was liver transplantation donor. AtlasImage, a software specific to array, was used to analyze the result. The expression pattern of some genes identified by Atlas arrays hybridization was confirmed by reverse transcription polymerase chain reaction (RT-PCR) in 24 pairs of specimens and Northern blot of 4 pairs of specimens. RESULTS: The differential expression of cell cycle/growth regulator in hepatocellular carcinoma (HCC) showed a stronger tendency toward cell proliferation with more than 1.5-fold up-regulation of Cyclin C, ERK5, ERK6, E2F-3, TFDP-2 and CK4. The anti-apoptotic factors such as Akt-1 were up-regulated, whereas the promotive genes of apoptosis such as ABL2 were down-regulated. Among oncogene/tumors suppressors, SKY was down-regulated. Some genes such as Integrin beta 8, Integrin beta 7, DNA-PK, CSPCP, byglycan, Tenacin and DNA Topo were up-regulated. A number of genes, including LAR, MEK1, eps15, TDGF1, ARHGDIA were down-regulated. In general, expression of the cancer progression genes was up-regulated, while expression of anti-cancer progression genes was down-regulated. These differentially expressed genes tested with RT-PCR were in consistent with cDNA array findings. CONCLUSION: Investigation of these genes in HCC is helpful in disclosing molecular mechanism of pathogenesis and progression of HCC. For the first time few genes were discovered in HCC. Further study is required for the precise relationship between the altered genes and their correlation with the pathogenesis of HCC.

Profiling of differentially expressed genes in human gastric carcinoma by cDNA expression array.

AIM: To investigate the expression of cancer related genes in gastric carcinoma (GC) through the use of Atlas Human Cancer Array membranes with 588 well-characterized human genes related to cancer and tumor biology. METHODS: Hybridization of cDNA blotting membrane was performed with (32)P-labeled cDNA probes synthesized from RNA isolated from gastric carcinoma and adjacent noncancerous gastric epithelial tissue. AtlasImage, which is a software specific to array, was used to analyze the result. RESULTS: The differentially expression cell cycle/growth regulator in GC showed a stronger tendency toward cell proliferation with 2.7-fold up-regulation of CK1. The promoter genes of apoptosis were down-regulated, including caspase-8 precursor, caspase-9 and caspase-10. Among the oncogene/tumor suppressor genes, ABL2 was down-regulated. In addition, some genes were up-regulated, including matrix metalloproteinse 2(MMP-2), MMP-16(MT3-MMP), SKY, CD9 and semaphorin V. A number of genes were down-regulated, including neuroendocrine-dlg (NE-dlg), retinoic acid receptor gamma and tumor suppressor DCC colorectal. In general, The expression of the cancer progression genes were up-regulated, while the expression of anti-cancer progression genes were down-regulated. CONCLUSION: Investigation of these genes should help to disclose the molecular mechanism of the onset, progression and prognosis of GC. Several genes are reported herein to be altered in GC for the first time. The quick and high-throughout method of profiling gene expression by cDNA array provides us with an overview of key factors that may involved in GC, and may aid the study of GC carcinogenesis and provide molecular targets for diagnosis and therapy. The precise relationship between the altered genes and gastric carcinogenesis is a matter for further investigation.

Clinical short-term results of radiofrequency ablation in liver cancers.

AIM: To study local therapeutic efficacy, side effects, and complications of radiofrequency ablation (RFA), which is emerging as a new method for the treatment of patients with hepatocellular carcinoma (HCC) with cirrhosis or chronic hepatitis and metastatic liver cancer. METHODS: Thirty-six patients with primary and secondary liver cancers (21 with primary hepatocellular carcinoma, 12 with colorectal cancer liver metastases and 3 with other malignant liver metastases), which were considered not suitable for curative resection, were include in this study. They were treated either with RFA (RITA2000, Mountain View, California, USA) percutaneously (n=20) or intraoperatively (n=16).The procedures were performed using the ultrasound guidance. The quality of RFA were based on monitoring of equipments and subject feeling of the practitioners. Patients treated with RFA was followed according to clinical findings,radiographic images, and tumor markers. RESULTS: Thirty-six patients underwent RFA for 48 nodules. RFA was used to treat an average 1.3 lesions per patient, and the median size of treated lesions was 2.5 cm (range, 0.5-9 cm). The average hospital stay was 5.6 days overall (2.8 days for percutaneous cases and 7.9 days for open operations). Seven patients underwent a second RFA procedure (sequential ablations). Sixteen HCC patients with a high level of alpha fetoprotein (AFP) and 9 colorectal cancer liver metastases patients with a high level of serum carcinoembryonic antigen (CEA) have a great reduction benefited from RFA. Four (11.1 %) patients had complications: one skin burn; one postoperative hemorrhage; one cholecystitis and one hepatic abscess associated with percutaneous ablations of a large lesion. There were 4 deaths: 3 patients died from local and system diseases (1 at 7 month, 1 at 9 month, and 1 at 12 month), 1 patients died from cardiovascular shock, but no RFA-related death. At a median follow-up of 10 months (range, 1-24 months), 6 patients (16.7 %) had recurrences at an RFA site, and 20 patients (56.7 %) remained clinically free of disease. CONCLUSION: RF ablation appears to be an effective, safe, and relatively simple procedure for the treatment of unresectable liver cancers. The rate and severity of complications appear acceptable. However, further study is necessary to assess combination with other therapies, long-term recurrence rates and effect on overall survival.

Integrin gene expression profiles of human hepatocellular carcinoma.

AIM: To investigate gene expression profiles of integrin genes in hepatocellular carcinoma (HCC) through the usage of Atlas Human Cancer Array membranes, semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Northern blot. METHODS: Hybridization of cDNA array membrane was performed with alpha(32)P-labeled cDNA probes synthesized from RNA isolated from hepatocellular carcinoma and adjacent non-cirrhotic liver. AtlasImage, which is a software specific to array, was used to analyze the result. RT-PCR of 24 pairs specimen and Northern blot of 4 pairs specimen were used to confirm the expression pattern of some integrin genes identified by Atlas arrays hybridization. RESULTS: Among 588 genes spotted in membrane, 17 genes were related to integrin. Four genes were up-regulated, such as integrin alpha8, beta1, beta7 and beta8 in HCC. Whereas there were no genes down-regulated in HCC. RT-PCR and Northern blot analysis of integrin beta1 gene gave results consistent with cDNA array findings. CONCLUSION: Investigation of these integrin genes should help to disclose the molecular mechanism of the cell adhesion, invasive and metastasis of HCC. A few genes are reported to have changed in HCC for the first time. The quick and high-throughout method of profiling gene expression by cDNA array provides us overview of key factors that may involved in HCC, and may find the clue of the study of HCC metastasis and molecular targets of anti-metastasis therapy. The precise relationship between the altered genes and HCC is a matter of further investigation.

Arterial chemotherapy of 5-fluorouracil and mitomycin C in the treatment of liver metastases of colorectal cancer.

AIM: Regional chemotherapy using hepatic artery catheters is a good method of treating patients with colorectal cancer liver metastases. We investigated the survival of patients with liver metastases from colorectal cancer using 5-fluorouracil (5-FU) and mitomycin C Cthrough implantable hepatic arterial infusion port. METHODS: Seventy-five patients with inoperable liver metastases from colorectal cancer were included between March, 1992 and November, 2001. We placed implantable hepatic arterial catheter (HAC) port by laparotomy. 5-FU, 1 000 mg/ m(2)/d continuous infusion for five days every four weeks, was delivered in the hepatic arterial catheter through the port. Mitomycin C, 30 mg/m(2)/d infusion in the first day every cycle through the port. Response to the treatment was evaluated by serial determinations of plasma CEA and imaging techniques consisting of computerized tomography and sonography of liver. RESULTS: Sixty-eight were performed hepatic artery chemotherapy and fifty-six were followed up among seventy-five HAC patients. Twenty-six patients(46.4 %) have responded and 4 complete remission were achieved. Eight patients (14.3 %) had stable liver metastases. Twenty-two patients (39.3 %) were progressed with increased tumor size and number. Twenty-nine patients(51.8%) had a decreased serum CEA level, while 10 patients (17.9 %) were stable and 17 patients (30.4 %) had an increased serum CEA level. There were no operative death in this series. Complications, which occurred in 18 patients (32.1 %), were as followed: hepatic artery thrombosis in 11, Upper gastric and intestinal bleeding in 3, liver abscess in 1, pocket infection in 1, cholangitis in 1, and hepatic artery pseudo-aneurysm in one patient. CONCLUSION: Combined infusion of 5-FU and mitomycin C by hepatic artery catheter port is an effective treatment for liver metastases from colorectal cancer. The high response and lower complication rates prove the adjuvant treatment of colorectal cancer with this treatment.

Gene expression profiles of hepatoma cell line BEL-7402.

BACKGROUND/AIMS: To investigate the gene expression of cancer related genes in hepatoma cell line BEL-7402 through the usage of Atlas Human Cancer Array with 588 well-characterized human genes related with cancer biology. METHODOLOGY: Hybridization of cDNA membrane was performed with 32P-labeled cDNA probes synthesized from RNA isolated from Human hepatoma cell line BEL-7402 and non-cirrhotic normal liver which was liver transplantation donor. Bioinformatics was used to analyze the result. The reverse transcription polymerase chain reaction of 24 pairs of specimens and northern blot of 4 pairs of specimens were used to confirm the expression pattern of some genes identified by Atlas arrays hybridization. RESULTS: The differential expression cell cycle/growth regulator in hepatoma cell line BEL-7402 showed a stronger tendency toward cell proliferation with up-regulation of E2F-3 and TFDP-2. The anti-apoptotic factors such as Akt-1 were up-regulated. Whereas the promotive genes of apoptosis were down-regulated, such as BAK and caspase-3. Besides this, some genes were up-regulated, such as Integrin beta 8, DNA-PK, CSPCP, cyclin C etc. A number of genes were down-regulated, which included LAR, ABL2, SKY, TDGF1 etc. In general, expression of the cancer progression genes were up-regulated, while expression of anti-cancer progression genes were down-regulated. The results of reverse transcription polymerase chain reaction of 24 pairs of specimens and Northern Blot of 4 pairs of specimens were consistent with the expression pattern of some genes identified by Atals arrays hybridization. CONCLUSIONS: Investigation of gene expression profile of hepatoma cell line BEL-7402 should help to disclose the molecular mechanism of the onset, progression and prognosis of hepatocellular carcinoma. The quick and high-throughout method of profiling gene expression by cDNA array provides us with an overview of key factors that may be involved in hepatocellular carcinoma, and may find the clue to the study of hepatocellular carcinoma carcinogenesis and molecular targets of diagnosis and therapy.

[Morphology of portal hypertension at the early stage of liver damage induced by CCl4: an experimental study with dogs].

OBJECTIVE: To explore the morphological basis of portal hypertension happened at the early stage of liver damage. METHODS: Sixteen mongrel dogs were injected intraperitoneally with carbon tetrachloride (CCl(4)) once a week for 4 weeks and then once every 4 days, totally for 7 weeks. Before the experiment and 7 weeks after the beginning of experiment, small pieces of liver tissues were taken through a median upper abdominal incision and underwent HE staining, Masson's staining, and immunohistochemistry with alpha-smooth muscle actin (alpha-SMA) to label the activated astrocyte (hepatic stellate cell, HSC), and transmission electron microscopy. The portal vein pressure was measured by puncture of gastroepiploic vein. The diameter of spleen was measure too. RESULTS: Since the second weeks after the experiment, the dogs showed vomiting, diarrhea, anorexia, weight loss, and varicose vein in the abdominal wall. One dog died of toxic enteritis 3 weeks after the beginning of experiment. 7 weeks after the injection of CCl(4), the average portal vein pressure of the rest 115 dogs was 22.7 cm H(2)O +/- 1.5 cm H(2)O, significantly higher than that before the experiment (12.2 cm H(2)O +/- 1.9 cm H(2)O, 1cm H(2)O = 0.098 kPa, P < 0.05). The long diameter of spleen of the 15 dogs was 24.4 +/- 3.1 cm, significantly greater than that before the experiment (18.7 cm +/- 2.4 cm, P < 0.05). Seven weeks after the injection of CCl(4) the appearance of liver did not change remarkably among the dogs, however, the spleens of the dogs showed swelling and stagnation of blood. No obvious ascites was seen. HE staining showed slight liver damage, such as fatty degeneration of liver cells; Masson's staining and electron microscopy showed typical sinusoid capillarization. alpha-SMA immunohistochemistry stain showed increased number of activated HSCs. Immunohistochemistry showed a number of alpha-SMA positive cells in the hepatic lobules. Portal hypertension was successfully induced in 15 dogs. CONCLUSION: At the early stage of liver damage the possible cause of portal hypertension is the increased resistance of blood stream due to sinusoid capillarization and activation of HSC.

A novel mechanism of abnormal hematological indices in liver cirrhosis: bone marrow endothelial cell dysfunction caused by humoral inhibitor affects the hematopoietic function of bone marrow.

Abnormal hematological indices (HIs), a complication of liver cirrhosis (LC), present difficulties in the treatment of LC and pose a serious threat to the survival of patients. LC is a dynamic wound-healing process that occurs in response to repeated liver injury and is a chronic disorder associated with changes in various organs and tissues. It has been reported that humoral inhibitor in the formation of LC could affect the hematogenic functions of bone marrow (BM) by acting on erythroid differentiation. This indicates that the BM microenvironment is affected by humoral inhibitor in LC. Bone marrow endothelial cells (BMECs) are very important components of the BM microenvironment that function as the cytoskeleton to support the adhesion of hematopoietic stem cells (HSCs). In addition, they can secrete cytokines, which have important functions in regulating positioning, homing, proliferation, differentiation and other functions of HSCs on the BM microenvironment. These functions of BMECs may be affected due to direct contact with blood and long-term exposure to an environment with humoral inhibitor in the presence of LC. Multiple studies have shown that during the formation of LC, hepatic sinusoid endothelial cells were damaged and secreted cytokines and matrix proteins. Moreover, these cytokines and matrix proteins were involved in the formation and development of LC. Similar in function to mature-stage BM, liver at the embryonic stage also functions as a type of hematogenic organ. With similar anatomical position and functions to that of hepatic sinusoid endothelial cells, BMECs may undergo similar changes and impair hematogenic function of BM. More importantly, we found even more convincing evidence in that the humoral inhibitor in LC could lead to the ultrastructural damage of BMECs that were positively related to the degree of severity of LC. Therefore, we hypothesise the existence of a novel mechanism for abnormal HIs in LC: the continuous humoral inhibitor may lead to abnormal cytokine secretion of BMECs and attenuate their supporting functions, and such alterations of BMECs may lead to BM microenvironment disorder and dysfunction of HSCs, finally causing abnormal HIs.

Serum bilirubin level is negatively correlated with disease progression of peripheral arterial disease: an observational cohort study.

We determined whether low bilirubin level is a risk factor for peripheral arterial disease (PAD). We recruited 318 patients with PAD and 100 healthy volunteers. Patients were divided into 4 groups by the Fontaine classification for PAD, namely, group 1 (grade 1, n = 4); group 2 (grade 2, n = 114), group 3 (grade 3, n = 164), and group 4 (grade 4, n = 36). Total bilirubin (T-BIL), direct bilirubin (D-BIL), and indirect bilirubin (I-BIL) levels were compared using stepwise multiple regressions adjusted for selected factors. After adjusting for gender, age, smoking, and diastolic blood pressure, serum levels of T-BIL, D-BIL, and I-BIL were significantly lower in the PAD group (P < .05). Patients with grade 4 PAD showed significantly (P < .05) lower levels of T-BIL when compared with grade 2 patients. We concluded that serum bilirubin levels are negatively correlated with the severity and progression of PAD.

[Effect of electroacupuncture on myoelectric activity of Jejunal limb After Roux-en-Y esophagojejunostomy].

OBJECTIVE: To explore the effect of electroacupuncture on myoelectric activity of Jejunal limb after Roux-en-Y esophagojejunostomy. METHODS: Fourteen health young pigs were randomly divided into 2 groups, an experimental group (total gastrectomy and Roux-en-Y esophagojejunostomy was carried out) and a control group (the abdominal cavity was closed after the electrode was placed), 7 pigs in each group. Electroacupunture was given at "Zusanli" (ST 36) in the experimental group. The changes of myoelectrogram of the jejunal limb was investigated. RESULTS: Compared with the control group, the amplitude and the frequency of the slow wave, and the amplitude and incidence rate of the spike potential in the experimental group were changed significantly; the duration of migrating motor complex (MMC) phase III was (2.6 +/- 0.7) minutes in the experimental group, which was significantly shorter than (7.1 +/- 1.1) minutes in the control group. Electroacupuncture did not significantly influence the amplitude and the frequency of the slow wave, but could increased significantly the incidence rate and the amplitude of the spike potential; after electroacupuncture, the duration of MMC phase III was (5.7 +/- 0.9) minutes, which was significantly longer than (2.6 +/- 0.7) minutes before electroacupuncture. CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) can relieve the Roux-en-Y stasis syndrome through influencing myoelectric activity of the jejunum.

[Surgical treatment of acute obstructing colorectal cancer: a report of 297 cases].

OBJECTIVE: To evaluate surgical treatment of obstructing colorectal cancer. METHODS: From July 1993 to July 2003, clinical data of 297 cases undergoing emergency operation for obstructing colorectal cancer were analyzed retrospectively. There were 103 cases with right-sided lesion and 194 cases with left- sided lesion. RESULTS: All patients received emergency operation. Stage i tumor resection and anastomosis were performed in 126 patients including 98 cases with right- sided lesion and 28 with left- sided lesion, total or subtotal colectomy in 108,Hartmann operation in 36,Dixon operation in 9, ileocolic anastomosis in 11,and colostomy in 7 cases. Postoperative complications occurred in 53 cases (17.8% ) including incision infection, intraperitoneal infection and intestinal fistula. There were 17 perioperative deaths. Two hundred and eighty cases healed (94.3% ). CONCLUSION: Stage i tumor resection and anastomosis and total or subtotal colectomy are feasible and safe surgical procedures for obstructing colorectal cancers.