Search

AIM To evaluate the prognostic value of the number of retrieved lymph nodes (LNs) and other prognostic factors for patients with distal cholangiocarcinomas, and to determine the optimal retrieved LNs cut-off number. METHODS The Surveillance, Epidemiology and End Results database was used to screen for patients with distal cholangiocarcinoma. Patients with different numbers of retrieved LNs were divided into three groups by the X-tile program. X-tile from Yale University is a useful tool for outcome-based cut-point optimization. The Kaplan-Meier method and Cox regression analysis were utilized for survival analysis. RESULTS A total of 449 patients with distal cholangiocarcinoma met the inclusion criteria. The Kaplan-Meier survival analysis for all patients and for N1 patients revealed no significant differences among patients with different retrieved LN counts in terms of overall and cancer-specific survival. In patients with node-negative distal cholangiocarcinoma, patients with four to nine retrieved LNs had a significantly better overall (P = 0.026) and cancer-specific survival (P = 0.039) than others. In the subsequent multivariate analysis, the number of retrieved LNs was evaluated to be independently associated with survival. Additionally, patients with four to nine retrieved LNs had a significantly lower overall mortality risk [hazard ratio (HR) = 0.39; 95% confidence interval (CI): 0.20-0.74] and cancer cause-specific mortality risk (HR = 0.32; 95%CI: 0.15-0.66) than other patients. Additionally, stratified survival analyses showed persistently better overall and cancer-specific survival when retrieving four to nine LNs in patients with any T stage of tumor, a tumor between 20 and 50 mm in diameter, or a poorly differentiated or undifferentiated tumor, and in patients who were ≤ 70-years-old. CONCLUSION The number of retrieved LNs was an important independent prognostic factor for patients with node-negative distal cholangiocarcinoma. Additionally, patients with four to nine retrieved LNs had better overall and cancer-specific survival rates than others, but the reason and mechanism were unclear. This conclusion should be validated in future studies.

// Kun He 1 , Xiwen Zhu 1 , Yan Liu 4 , Chunmu Miao 1 , Tao Wang 1 , Peizhi Li 1 , Lei Zhao 2 , Yaxi Chen 2 , Junhua Gong 1 , Can Cai 4 , Jinzheng Li 1 , Shengwei Li 1 , Xiong Z. Ruan 2, 3 , Jianping Gong 1 1 Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China 2 Centre for Lipid Research, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China 3 Centre for Nephrology, University College London (UCL) Medical School, Royal Free Campus, London, United Kingdom 4 Department of Gastroenterology and Hepatology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China Correspondence to: Jianping Gong, email: gongjianping11@126.com Shengwei Li, email: lishengwei11@163.com Keywords: NLRP3 inflammasome, Kupffer cell, non-alcoholic fatty liver disease, IL-1β Received: March 20, 2017 Accepted: April 17, 2017 Published: April 27, 2017 ABSTRACT NOD-like receptor (NLR) NLRP3 inflammasome activation has been implicated in the progression of non-alcoholic fatty liver disease (NAFLD) from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH). It has been also shown that palmitic acid (PA) activates NLRP3 inflammasome and promotes interleukin-1β (IL-1β) secretion in Kupffer cells (KCs). However, the specific mechanism of the NLRP3 inflammasome activation is unclear. We studies the molecular mechanisms by investigating the roles of Thioredoxin-interacting protein (TXNIP) and NLRP3 on NAFLD development in patients, high-fat diet (HFD)-induced NAFL and methionine choline deficient (MCD) diet-induced NASH in wild type (WT), TXNIP −/− (thioredoxin-interacting protein) and NLRP3 −/− mice, and isolated KCs. We found that the expressions of NLRP3 and TXNIP in human liver tissues were higher in NASH group than in NAFL group. Furthermore, co-immunoprecipitation analyses show that activation of the TXNIP-NLRP3 inflammasome protein complex occurred in KCs of NASH WT mice rather than NAFL WT mice, thus suggesting that the formation and activation of this protein complex is mainly involved in the development of NASH. NLRP3 −/− mice exhibited less severe NASH than WT mice in MCD diet model, whereas TXNIP deficiency enhanced NLRP3 inflammasome activation and exacerbated liver injury. PA triggered the activation and co-localization of the NLRP3 inflammasome protein complex in KCs isolated from WT and TXNIP −/− but not NLRP3 −/− mice, and most of the complex co-localized with mitochondria of KCs following PA stimulation. Taken together, our novel findings indicate that TXNIP plays a protective and anti-inflammatory role in the development of NAFLD through binding and suppressing NLRP3.

FcγRIIIa (CD16) is a low-affinity Fc receptor of IgG. As the idio-binding receptor of IgG Fc, it plays an important role in the antibody-dependent cellular cytotoxicity of natural killer cells. The aim of the present study was to investigate the distribution of Kupffer cells (KCs) and the expression of their surface receptor FcγRIIIa in hepatocellular carcinoma. Furthermore, we also aimed to observe the functional mechanism of FcγRIIIa. Immunohistochemical analysis was employed to study KCs and FcγRIIIa. In order to explore the role of FcγRIIIa in the growth of cancer cells, KCs and H22 tumor cells were co-cultured in different serum. The mRNA expression levels of tumor necrosis factor (TNF)-α and FcγRIIIa were analyzed by RT-qPCR; the TNF-α and FcγRIIIa protein expression levels were examined by enzyme‑linked immunosorbent assay and western blot analysis, respectively. Our results showed that the number of Kuppfer cells in cancerous tissues (21.6±7.8) was lower than those in para-cancerous (68.8±9.1) tissues and adjacent normal hepatic tissues (62.0±1.9) (P

Systemic inflammation affects hemostasis during severe acute pancreatitis (SAP). A hypercoagulable state occurs more frequently in SAP, which is not fully detected by traditional coagulation testing.The aim of this study was to evaluate the contribution of clot formation and lysis (CloFAL) assay to improve monitoring of global coagulation in patients with SAP.Twenty-five patients with SAP who were treated from December 2009 to April 2011 were studied. Plasma was collected at the time of admission, and CloFAL was measured using the CloFAL analyzer. The parameters evaluated include coagulation time (CT), fibrinolysis time (FT), and maximum amplitude (MA), from which the accelerating coagulation extent (ACE, MA/CT), accelerating fibrinolytic extent (AFE, MA/FT), and balance level exponent (BLE, ACE/AFE) were calculated. In addition, laboratory values for the traditional coagulation testing were measured. Values were compared to a control group of 20 healthy subjects.The MA, FT, ACE, and BLE values of the CloFAL assay were significantly increased in the SAP group compared to the control group (p\0.05 for all measurements). For the traditional coagulation testing, fibrinogen, plasminogen, and D-dimer levels were higher in patients in the SAP group compared to the control group (p\0.05).Our findings using the CloFAL analyzer indicate that the hypercoagulable state was due to increased fibrin generation and invariable fibrinolysis in patients with SAP. CloFAL assay is a simple and useful global coagulation assay to monitor hypercoagulable states during SAP.

Since the widespread adoption of laparoscopic cholecystectomy (LC) in the late 1980s, a rise in common bile duct (CBD) injury has been reported. We analyzed the factors contributing to a record of zero CBD injuries in 10 000 consecutive LCs.The retrospective investigation included 10 000 patients who underwent LC from July 1992 to June 2007. LC was performed by 4 teams of surgeons. The chief main surgeon of each team has had over 10 years of experience in hepatobiliary surgery. Calot's triangle was carefully dissected, and the relationship of the cystic duct to the CBD and common hepatic duct was clearly identified. A clip was applied to the cystic duct at the neck of the gallbladder and the duct was incised with scissors proximal to the clip. The cystic artery was dissected by the same method. Then, the gallbladder was dissected from its liver bed. A drain was routinely left at the gallbladder bed for 1-2 days postoperatively.No CBD injuries occurred in 10 000 consecutive LCs, and there were 16 duct leaks (0.16%). Among these, there were 10 Luschka duct leaks (0.1%) and 6 cystic duct leaks (0.06%). Four hundred thirty cases were converted to open cholecystectomy (OC), giving a conversion rate of 4.3%. After a mean follow-up of 17.5 months (range 6-24 months), no postoperative death due to LC occurred, and good results were observed in 95% of the patients.In our 10 000 LCs with zero CBD injuries, the techniques used and practices at our department have been successful. Surgeon's expertise in biliary surgery, preoperative imaging, precise operative procedures, and conversion from LC to OC when needed are important measures to prevent CBD injuries.

To investigate the feasibility of repairing bone defect with methods of tissue-engineering and human bone morphogenetic protein-2 (hBMP-2) gene transfection in osteoporotic rats.Twenty-four 6-month-old female Sprague-Dawley rats underwent ovariectomy, while 8 rats received sham-operations. Three months later, bone mesenchymal stem cells (BMSC) harvested from osteoporotic rats were divided into two groups randomly. Experimental group were transfected by recombinant plasmid carrying hBMP-2 gene, and control group left untreated. All BMSC were seeded into coralhydroxyapatite scaffolds. Then the cell/scaffold constructs were implanted into the defect site created in the ramus of mandible of osteoporotic rats respectively.Positive results were confirmed by immunohistochemistry and in situ hybridization in experimental group. New bone formation was found at the margin of the defect treated with the BMSC modified by hBMP-2 gene transfer at 4 weeks after implantation and appeared mature 8 weeks after the treatment. However, the amount of newly formed bone was much less and there was some adipose tissue at defect margins 8 weeks after implantation in control group.The results of this experiment indicate that BMSC-mediated rhBMP-2 gene therapy in conjunction with bone tissue engineering may allow for successful treatment of large bone defects in osteoporosis rats.

To determine the causes and incidence of facial injuries by an epidemiologic retrospective study.A total of 3 958 patients with facial injuries treated at Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University from 1955 to 2001 were investigated. Data regarding age, gender, cause of injury, pattern of fracture and associated systemic injuries were reviewed.The male to female ratio of the patients with facial injury was 4.27:1 and 33.4% of patients were aged between 21 and 30 years. The most common cause of injury was traffic accident (30.6%), followed by falls (21.4%) and collision (15.8%). A total of 794 patients (20.1%) showed only soft tissue injuries. 1 100 patients (27.8%) had multiple fractures in facial bones and 2,064 patients (52.1%) had single fracture. The mandibular fracture was most frequently seen, followed by the maxilla and the zygoma. The most common site of mandible fracture was the body (31.2%), followed by the symphysis (22.7%), the condylar (20.5%) and the angle (13.7%). Accompanied injuries to brain and skull happened in 916 patients (23.1%).Bone fractures were more common in hospitalized patients with facial injuries. The numbers and sites of fracture were related to the causes of injuries and anatomic structure of the bone. The brain and skull injuries, the most often and seriously accompanied injuries, would not be neglected.

Van der Woude syndrome (VWS) (OMIM 119300) is a dominantly inherited, developmental disorder that is characterized by pits and/or sinuses of the lower lip and a cleft lip and/or cleft palate. Mutations in the interferon regulatory factor 6 gene ( IRF6) have been recently identified in patients with VWS, with more than 60 mutations reported. However, the VWS phenotype, IRF6 mutation genotypes, and their interrelationships in Chinese VWS patients have not been studied. Here, we report 11 Chinese families with variable clinical phenotypes of VWS and identified mutations in all patients. Of the 11 mutations, 8 appeared to be novel: CC5.6GT, T342A, 566delA, C748T, C756A, C989A, C1209G, and 1316delT. Seven mutations caused a change or loss of the IRF6 domain. The marked phenotypic variation may be caused by the action of certain modifier genes on IRF6 function. Abbreviations: VWS, Van der Woude syndrome; IRF6, interferon regulatory factor 6; CL/P, cleft lip and/or cleft palate; DBD, DNA-binding domain; SMIR, Smad-interferon regulatory factor-binding domain; Kb, kilobase; PCR, polymerase chain-reaction.

To study the biological features and osteoblast/adipocyte phenotypes of bone marrow stromal cells (BMSCs) of Sprague-Dawley (SD) rats with Type I osteoporosis by induced culture.Six-month-old SD rats were used in this study. 16 female rats were randomly divided into two groups. Eight rats were ovariectomied as experimental group to establish the modle of Type I osteoporosis, while other rats received sham-operation. Three month later BMSCs of 16 rats were isolated by discontinueous gradient centrifugation and then plated in alpha-MEM medium as primary culture. Secondary harvested cells were cultured for 14 days in alpha-MEM medium supplemented with dexamethasone, ascorbic acid, vitamin D3, beta-glycerophosphate or dexamethasone, 3-isobutyl-1-methylxanthine, insuline, and indomethine. The cells were screened by inverted microscope each day and cell growth was studied with cell counting. The osteoblast and adipocyte phenotypes were verified by cytochemistry staining, counted the percentage of positive stained cells.The weight and bone mineral density of rats were statistically different between experimental group and control group. Gomori and Von Kossa's staining demonstrated positive osteoblast phenotypes of alkaline phosphatase and mineralized nods by osteogenic inducer, while Oil Red O staining identified BMSCs treated with adipogenic medium resulted in adipocyte formation and there was no significant difference in the percentage of positive stained cells between two groups.The model of Type I osteoporosis has been established successfully. BMSCs from SD rats with osteoporosis maintain their differentiation potential.

This experiment was designed to develop an experimental model of mandibular distraction osteogenesis in goats for determining the principle and the process of distraction osteogenesis.Eight 3-month-old, healthy domestic goats were randomly divided into four groups (n = 2 per group). The animals from each group were killed at 1, 2, 4, and 6 weeks after bone distraction, respectively. The sections of newly formed bone were obtained and evaluated with histologic and radiographic study. We also assessed the newly formed bone with bone density examination.The specimens of right mandible increased averagely 10 mm after distraction. The Newly formed bone was demonstrated histologically. The X-ray examination and bone density measurements showed that the bone density increased with the time of fixation.Goat is a proper experimental animal for modeling mandibular distraction osteogenesis, and in this regal, it is important to keep the direction of distracter force in conformity with the direction of the expected distraction osteogenesis for avoiding the side force and thus ensuring successful distraction and good quality of newly formed bone.

To make sure whether super-high molecular weight polylactate is toxic to the body after it has been implanted into the body for a long period.We implanted super-high molecular weight polylactate into the rats and took the specimens of blood at 3, 6, 9, 12 months after the operation. The changes of proteins, electrolyte, enzyme and other indices were observed. At the same time, the tissue around the implants were taken out to carry out the histological observation.At 3, 6, 9, 12 months after the operation, the levels of albumin, globulin, total bilirubin, direct bilirubin, triglyceride, glucose, K+, Na+, Cl-, Ca2+, alkaline phosphatase, glutamic-pyruvic transaminase in the blood plasma were all in the normal range; there were no significant differences between the experimental group and the control group. The level of lactate dehydrogenase increased slightly, but there was no statistically significant difference between the experimental group and the control group. There were no non-reversible immune rejection around the implants in the histological observation.Super-high molecular weight polylactate is not toxic to the body after it has been implanted into the animals for a long period.

Pancreatic encephalopathy (PE), an unfamiliar complication of severe acute pancreatitis (SAP), is difficult to diagnose and treat, and it has a high mortality. The aim of this study was to investigate the manifestation, classification, mechanism and therapy of PE.Of 132 patients with SAP treated at our hospital from March 1994 to March 2004, 24 patients complicated by PE were analyzed retrospectively.The causes of SAP were mostly biliary and alcoholic. Twenty-four patients (18.2%) were complicated by PE within 3 hours-38 days(average 6.6 days)[21(87.5%) within 2 weeks, and 3(12.5%) after 2 weeks] . Eleven patients were male and 13 female, with an average of 47 years (range 25-72 years). Excitement or restrain was the main manifestation. Nine patients (37.5%) received surgery and 15(62.5%) conservative treatment, with a mortality of 11.1%(1/9) and 66.7%(10/15), respectively.PE occurs 2 weeks after SAP and is part of multiple organ failure (MOF). Some patients have PE in the late stage of SAP because of lack of VitB1 and nutrition. But PE can be prevented by prescribing adequate nutrition and VitB1 in the early stage of SAP.

To isolate and chondro-inductive culture of human adipose tissue-derived stromal cells and to study their heterotopic chondrogenesis by loading them on alginate gel.Liposuction human adipose tissues were minced and digested with collagenase type I. The obtained stromal cells were primarily cultured in BGJb medium for ten days. Secondary harvested cells were cultured in DMEM-F12 medium supplemented with 10%FBS, 6.25 mg/L insulin, 10 mg/L TGF-beta1, 50 mg/L of freshly prepared L-ascorbate for 14 days. After in vitro assay of chondrogenic phenotypes, the cells at density of 10(10)/L were mixed with 1.2% alginate sodium and 102 mmol/L CaCl(2). The cross-linking cell-alginate gel were injected into four BALB/C athymic mice subcutaneously (1 ml for each mouse). Meanwhile, the auto-controls were set by injecting equal dose of simple alginate gel and pure cells in two opposite buttocks of the same mouse subcutaneously. Two mice were sacrificed at fourth and eighth week postoperatively and all samples were removed, fixed, embedded in paraffin and cut into sections of 5 micro m thick. HE staining, Alcian blue and modified Masson's trichrome staining were employed to observe chondrogenesis histologically.Alcian blue and immunocytochemical staining revealed chondroitin sulfate and collagen II in cell matrix after having been chondro-inductive cultured for 14 days. At intervals of fourth and eighth week, heterotopic chondrogenesis is (cartilage formed) within cell-alginate injected sites were found in all mice but negatively in auto-controls. Histologically the hypertrophic chondrocytes were among cartilage matrix in different staining. All alginate gel and solitory cells absorbed within two to three weeks postoperatively in auto-controls.It seems that stromal cells derived from human adipose tissue presents a potential for chondrogenic differentiation.

To elucidate the functional status of dendritic cells (DC) in the tissue of oral squamous cell carcinoma by analyzing characteristic phenotype of them.34 specimens from oral squamous cell carcinoma cases primarily treated with surgery were selected as test group. In addition, 30 specimens of normal mucosa from oral mucocele cases were used as control. Distribution of DC expressing CD1a+, HLA-DR+ and CD83+ in tumor tissue and normal mucous membrane was observed by immunohistochemistry. The number of DC expressing the antigens, which represented the density of DC infiltrating into tissue, was counted by microscope. The density of DC and the rate of DC expressing HLA-DR in oral carcinoma group and control were statistically compared.There was no CD83+ DC in all cases, but CD1a+ DC was found in all samples. The density of CD1a+ DC in tumor tissue was significantly lower than that in normal mucous membrane (P0.05). HLA-DR antigen expressed on the surface of DC in tumoral epithelium of 27-case carcinoma specimens and in normal mucous epithelium of 23 cases. The rate of HLA-DR positive expression of TIDC had no statistic significance between the two groups.The lower density of DC infiltrating in tumor tissue might reflect the microenviromental immunodeficiency of hosts with oral squamous cell carcinoma, and the functional mature of DC might be inhibited by the immunosuppressive action of oral squamous cell carcinoma.

To study the differential expression of transforming growth factor beta 1 (TGF beta 1) in oral carcinoma and stroma lymphocytes by induction chemotherapy and inquire into the mechanism of TGF beta 1 information transmission.Forty cases of oral tumor were treated with MTX, CDDP and PYM via subcutaneous implantable drug pump, in situ hybridization method was adopted to detect the expression of TGF beta 1 mRNA.The positive expression of TGF beta 1 mRNA was enhanced in oral carcinoma (P0.05). After the induction chemotherapy via subcutaneous implantable drug pump, not only the expression level of TGF beta 1 in malignant cells of invading front zone was up-regulated (P0.05), but the expression level of stroma lymphocytes was higher than before.These data demonstrated that TGF beta 1 not only has transforming potential, but also enhances the malignant progression of oral carcinoma. It was clear that TGF beta 1 can act as a tumor suppressor and a significant stimulator of T-cell-mediated tumor cytotoxity as well.

To explore the changes of plasma endothelin (ET) and nitric oxide (NO) levels in patients with acute pancreatitis.The level of plasma ET was measured by radioactive-immunoassay, and NO by spectrophotometry.The levels of ET, NO and the ET/NO ratio in patients with severe acute pancreatitis (SAP) within 24 hours in hospital were all significantly higher than those in other groups of patients [(176+/-8) pg/ml, (97+/-11) micromol/L, and 1.83+/-0.12, P0.01]. Compared to healthy controls (N), the levels of ET and NO in patients without pancreatitis acute abdomen (NAP) and patients with mild acute pancreatitis (MAP) increased significantly (P0.01). After appropriate treatment, the levels of ET and NO in the MAP group were lower (P0.01). Compared with those before treatment, the levels of ET and NO in the SAP group on the 3rd and 7th day in hospital dropped significantly (P0.01). The ET/NO ratio on the 7th day was also lower than that on admission (P0.01).The malfunction of endothelial cells and the increased ET/NO ratio may be related to the mechanism of pancreatic microcirculatory disturbance in patients with SAP; early dynamic determination of these parameters may help predict the prognosis of SAP.

To determine the role of mononuclear macrophages in the pathogenesis of acute lung injury during acute obstructive cholangitis.Sixty Wistar rats were used to study the correlation between the behavior of mononuclear macrophages and acute pulmonary injury during acute obstructive cholangitis (AOC). Animal model of AOC was made according to the method that the common bile duct was injected with Escherichia coli and ligated. The rats were killed at 6 h, 12 h, 24 h and 48 h after operation. The phagocytic function of Kupffer cells (KCs), the number of alveolar macrophages (AMs) in bronchoalveolar lavage liquid, and the extravascular water content of lung tissue were measured. The levels of lipid peroxide (LPO) and supperoxide dismutase (SOD) were determined too. Pathological alterations of liver and lung tissue were observed under light and electron microscopes.KCs phagocytic function was significantly elevated at the 6th hour but markedly decreased from the 24th hour to the 48th hour in the AOC group as compared with the control (P0.05). From the 12th to the 48th hour, the number of AMs, the extravascular water content of lung tissue, and the content of LPO significantly increased, but the SOD level of lung tissue decreased greatly (P0.05). Morphologically, KCs proliferated diffusely in the early period in livers of the AOC group, but decreased markedly in the late period. Mitochondria of KCs were swollen or even vacuolated; focal cytoplasmic degeneration and many myeli like figures could be seen in the cytoplasm. The changes of injury such as disturbance of pulmonary capillary blood circulation, degeneration and/or necrosis of the lung tissue and endothelium, and inflammatory reactions could be observed. In other two groups, no evident morphological changes were observed.KCs phagocytic function is decreased, whereas AM is activated by the invading bacteria to release such inflammatory mediators as free radicals, resulting in acute pulmonary injury. It seems that there is a close relationship between the functional status of mononuclear macrophages and the development of acute lung injury. The dysfunction of mononuclear macrophages may play an important role in the pathogenesis of multiple organ damage, especially acute pulmonary injury.

Objective: To summarize the diagnosis and treatment of juxta-ampullary duodenal diverticulum (JAD) in our hospital. Methods: Of 5000 consecutive endoscopic retrograde cholangiopancreatography (ERCP) performed in our department, 225 patients were diagnosed with JAD and treated. All patients were classified based on the location of Ampullae of Vater in relation to the duodenal diverticulum. Of the 225 JAD patients, 96 patients (43%) required surgery. Results: The 225 patients with JAD were divided into Type A (146 cases, 65%) or Type B (79 cases, 35%). Type A patients presented with papillae near the diverticulum or in its margin. In this type, 36 patients (25%) presented with diverticulitis, bleeding, perforation or cholelithiasis, and were treated surgically. Type B patients presented with papillae inside the diverticulum. Among them, 60 patients (76%) had complications requiring surgery. Conclusions: JAD can be divided into two types based on location of the papillae. ERCP was the primary method of diagnosing JAD and patients with severe complications required surgical intervention.

AIM: To observe the synthesis of endotoxin receptor CD14 protein and its mRNA expression in Kupffer cells (KCs), and evaluate the role of CD14 in the pathogenesis of liver injury in rats with alcohol-induced liver disease (ALD). METHODS: Twenty-eight Wistar rats were divided into two groups: ethanol-fed group and control group. Ethanol-fed group was fed ethanol (dose of 5-12g·kg·d-1) and control group received dextrose instead of ethanol. Two groups were sacrificed at 4 wk and 8 wk, respectively. KCs were isolated and the synthesis of CD14 protein and its mRNA expression in KCs were determined by flow cytometric analysis (FCM) or the reverse transcription polymerase chain reaction (RT-PCR) analysis. The levels of plasma endotoxin and alanine transaminase (ALT) were measured by Limulus Amebocyte Lysate assay and standard enzymatic procedures respectively, and the levels of plasma tumor necosis factor (TNF)-α and interleukin (IL)-6 were both determined by ELISA. The liver pathology change was observed under light and electric microscopy. RESULTS: In ethanol-fed group, the percentages of FITC-CD14 positive cells were 76.23% and 89.42% at 4 wk and 8 wk, respectively. Compared with control group (4.45% and 5.38%), the difference was significant (P < 0.05). The expressions of CD14 mRNA were 7.56 ± 1.02 and 8.74 ± 1.37 at 4 wk and 8 wk, respectively, which were significantly higher compared with the control group (1.77 ± 0.21 and 1.98 ± 0.23) (P < 0.05). Plasma endotoxin levels at 4 wk and 8 wk increased significantly in ethanol-fed group (129 ± 21 ng·L-1 and 187 ± 35 ng·L-1) than those in control rats (48 ± 9 ng·L-1 and 53 ± 11 ng·L-1)(P < 0.05). Mean values of plasma ALT levels increased dramatically in ethanol-fed rats (112 ± 15 IU/L and 147 ± 22 IU/L) than those in the control animals (31 ± 12 IU/L and 33 ± 9 IU/L) (P < 0.05). In ethanol-fed rats, the levels of TNF-α were 326 ± 42 ng·L-1 and 402 ± 51 ng·l-1 at 4 wk and 8 wk, respectively which were significantly higher than those in control group (86 ± 12 ng·L-1 and 97 ± 13 ng·L-1) (P < 0.05). The levels of IL-6 were 387 ± 46 ng·L- 1 and 413 ± 51 ng·L-1, which were also higher than control group (78 ± 11 ng·L-1 and 73 ± 10 ng·L-1) (P < 0.05). In liver section from ethanol-fed rats, there were marked pathological changes including steatosis, cell infiltration and necrosis. No marked pathological changes were seen in control group. CONCLUSION: Ethanol administration led to a significant synthesis of endotoxin receptor CD14 protein and its gene expression in KCs, which maybe result in the pathological changes of liver tissue and hepatic functional damages.

To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome.Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA).The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/-37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P0.05).NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome.