Effect of Echinococcus granulosus cyst fluid on the proliferation of HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether Echinococcus granulosus cyst fluid-infected host liver cells had differential expression of mitogen-activated protein kinases (MAPKs) or differential cell cycle activity.</p><p><b>METHODS</b>Human liver cells cultured with different concentrations of hydatid cyst fluid (HCF) were tested by the MTT method to determine effects on proliferation. The cell cycle was assessed by flow cytometry. Western blotting was used to detect changes in protein expressions of p-ERK, PCNA, cyclin-A, cyclin-B1, cyclin-D1, and cyclin-E.</p><p><b>RESULTS</b>Forty-eight, 72 and 96 h of HCF at 15%, 30% and 60% concentrations in the cell media significantly promoted cell proliferation (F=67.845, P less than 0.01) and compared to controls (P less than 0.05). Cells exposed to 15% HCF for 48 h showed significantly induced expression of p-ERK (F=1.916, P less than 0.01), higher than controls (P less than 0.01). Cells exposed to 15% HCF for 24 h showed significantly induced expression of cyclin-Dl (F=3.901, P less than 0.01), higher than controls (P less than 0.01). Cells exposed to 15% HCF for 48 h or 30% HCF for 72 h showed significantly induced expression of PCNA (F=91.140, P less than 0.01), higher than controls (P less than 0.01). Cells exposed to 15% HCF for 48 h or 30% HCF for 72 h shed significantly induced expression of cyclin-A (F=18.587, P=0.002), higher than controls (P less than 0.01). Cells exposed to 15% HCF for either 48 h or 72 h showed significantly induced expression of cyclin-B1 (F=2.064, P less than 0.01), higher than controls (P less than 0.01). Cells exposed to 30% HCF for 96 h showed significantly induced expression of cyclin-E (F=1.068, P less than 0.01), higher than controls (P less than 0.01).</p><p><b>CONCLUSION</b>Hydatid cyst fluid exerts no inhibitory effect on primary cultured host liver cells, but may promote cellular proliferation.</p>

Immune responses on allograft heart transplantation in inbred rats infected with Echinococcosis multilocularis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Alveolar echinococcosis (AE) is caused by the metacestode stage of Echinococcus multilocularis (E. multilocularis) and is a rare but life-threatening disease. This disease commonly is characterized by an infiltrative, tumor-like growth of the E. multilocularis metacestode in the liver of human. Liver transplantation is an effective therapy for end-stage of hepatic AE, but the characteristics of host immunity associated with E. multilocularis infection with organ transplantation are poorly defined. We hereby aimed to study the immunological status and allograft heart survival in inbred rats with E. multilocularis infection.</p><p><b>METHODS</b>Rat models of AE were established by injecting the E. multilocularis suspension made from E. multilocularis infected tissues into the abdomen of Lewis (LEW) rats. Three months later, in the experimental group, allograft heart transplantation was performed from Brown-Norway (BN) rats to the E. multilocularis infected LEW rats. In the control group, we transplanted hearts from BN rats to healthy LEW rats. The influence of the disturbed immune system in E. multilocularis infected rats on the heart transplantation was assessed, including observation of allograft heart survival time, histopathological examination of grafts and immunohistochemical examination of infiltrating cells (CD4(+) T cells, CD8(+) T cells and eosinophile granulocytes), measurement of interleukin (IL)-4 and interferon (IFN)-γ in the serum by enzyme-linked immunosorbent assay (ELISA) and analysis of CD4(+)CD25(+) regulatory T cells in peripheral blood by fluorescence activated cell sorting (FACS) flow cytometric analysis.</p><p><b>RESULTS</b>The survival time of recipients in the experimental group was prolonged compared with those in the control group. The numbers of graft infiltrating CD8(+) T cells were decreased whereas the graft infiltrating eosinophil granulocytes (CD15(+)) were increased in grafts in the experimental group (P < 0.05). Furthermore, the proportion of CD4(+)CD25(+) regulatory T cells in the peripheral blood was 10.8% on average in the experimental group, which was significantly higher than that in the control group (6.1%). In addition, the level of serum IL-4 in E. multilocularis infected rats was higher than that in the control group rats, whereas the level of serum IFN-γ in experimental group was lower than that in the control group when graft rejection occurred (P < 0.05).</p><p><b>CONCLUSIONS</b>This study suggests that E. multilocularis infection could prolong the allograft survival time through the polarization of Th1/Th2-type cells and induction of CD4(+)CD25(+) regulatory T cells. This strategy may provide a new idea for establishing transplantation tolerance.</p>

Application of a digital three-dimensional reconstruction technique in the diagnosis and treatment of hepatic alveolar echinococcosis / 中华消化外科杂志

ObjectiveTo investigate the value of a digital three-dimensional reconstruction technique in the treatment of hepatic alveolar echinococcosis (HAE).MethodsThe computed tomography scan data for 13 patients with HAE who were admitted to the First Affiliated Hospital of Xinjiang Medical University from February 2011 to October 2011 were reconstructed and analyzed by a three-dimensional reconstruction system to assess resectability,and to facilitate surgical planning and individualized virtual surgery.The results of preoperative analysis were compared with the results of actual operations.ResultsThe three-dimensional models of the liver were reconstructed successfully,and intrahepatie lesions and vessels were clearly displayed.One patient received an autologous liver transplantation,10 underwent hepatectomy,and 2 received percutaneous transhepatic cholangial drainage.Virtual operation planning was carried out for 11 patients using the three-dimensional reconstruction system.The mean volume of the liver to be resected was predicted to be 920 ml (range,339-2678 ml),and the mean percentage of liver to be resected to the total liver volume was predicted to be 45％ ( range,23％ -68％ ).The mean volume o[ the actual liver resection was 834 ml (range,315-2250 m[),and the mean percentage of actual liver resected to the total liver volume was 42％ (range,22％ -70％ ),which was consistent with the results of preoperative three-dimensional reconstruction.All patients were followed up for 2-8 months,and no severe complications such as liver failure,hemorrhage and bile leakage were detected.ConclusionDigital three-dimen-sional reconstruction is helpful in the diagnosis and treatment of HAE and effectively reduces surgical risks.

Suppression of acute rejective response following orthotopic liver transplantation in experimental rats infected with Echinococcus multilocularis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hepatic alveolar echinococcosis (AE) is a parasitic disease in humans and caused by the Echinococcus multilocularis (Em). Orthotopic liver transplantation (OLT) may be the only effective treatment for end-stage hepatic AE. However, in some AE patients, extrahepatic Em can not be completely eliminated after OLT. We aimed to study whether the immunological changes caused by Em evasion may influence the rejective response.</p><p><b>METHODS</b>Rat modles of AE were established by injecting the Em suspension into abdomen of Brown Norway (BN) rats. Three months later, in the experimental group, the liver was transplanted from Lewis (LEW) rats to Em-infected BN rats. In the control group, transplantation was from LEW rats to healthy BN rats. Liver tissue and peripheral blood (PB) samples were collected on days 1, 3, 5, and 7 after OLT. Liver tissue was analyzed after hematoxylin and eosin (H&E) staining; numbers of CD4, CD8, and CD28 on peripheral blood cells were detected by flow cytometry; and expression of the chemokine fractalkine (Fkn) was detected by reverse transcription PCR (RT-PCR). Interleukin-10 (IL-10) was measured in the serum by enzyme-linked immunosorbent assay (ELISA). In every group, eight BN rats were retained for observing survival time.</p><p><b>RESULTS</b>The survival times of recipients in the experimental group were prolonged compared with those in the control group. The rejective response occurred later and was milder in the experimental group. percentage of CD4, CD8, CD28 T-cells and Fkn mRNA expression were lower in the experimental group. While the serum IL-10 levels were higher in the experimental group than those in the control group.</p><p><b>CONCLUSIONS</b>Acute rejective response after OLT was attenuated in the rats with Em infection, and the recipients` survival time was prolonged. Em may play a role in this process by elevating IL-10 secretion, decreasing the effector T cells, inhibiting the expression of Fkn, which lead to reduce the inflammatory cells infiltration into the liver.</p>

Ex vivo liver resection followed by autotransplantation for end-stage hepatic alveolar echinococcosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>For patients with end-stage hepatic alveolar echinococcosis (AE), in vivo resection of the involved parts of the liver is usually very difficult, therefore, allogenic liver transplantation is indicated. However, we hypothesize that for selected patents, ex vivo liver resection for thorough elimination of the involved tissues and liver autotransplantation may offer a chance for clinical cure.</p><p><b>METHODS</b>We presented a 24-year-old women with a giant hepatic AE lesion who was treated with hepatectomy, ex vivo resection of the involved tissue and hepatic autotransplantation. The patient had moderate jaundice and advanced hepatic AE lesion which involved segments I, IV, V, VI, VII, VIII and retrohepatic inferior vena cava. The lateral segments (II and III) of the left liver remained normal with over 1000 ml in its volume. No extrahepatic metastases (such as to the lung or brain) could be found. As the first step of treatment, X-ray guided percutaneous transhepatic cholangiodrainage (PTCD) was performed twice for bile drainage in segment III and II separately until her serum total bilirubin decreased gradually from 236 to 88 µmol/L. Total liver resection was then performed, followed by extended right hepatic trisegmentectomy and the entire retrohepatic vena cava was surgically removed en bloc while her hemodynamics parameters were stable. Neither veino-veinous bypass nor temporary intracorporeal cavo-caval or porto-caval shunt was used during the 5.7-hour anhepatic phase. The remained AE-free lateral segments of the left liver were re-implanted in situ. The left hepatic vein was directly anastomosed end-to-end to the suprahepatic inferior vena cava due to the lack of the retrohepatic inferior vena cava with AE total infiltration. Because compensatory retroperitoneal porto-caval collateral circulation developed, we enclosed remained infrahepatic inferior vena cava at renal vein level without any haemodynamics problems.</p><p><b>RESULTS</b>During a 60-day following-up after operation, the patient had a good recovery except for a mildly elevated serum total bilirubin.</p><p><b>CONCLUSIONS</b>As a radical approach, ex vivo liver resection and liver autotransplantation in a case has shown a optimal potential for treatment of the end-stage hepatic AE. Strict compliance with its indications, evaluation of vessels of patients pre-operatively, and precise surgical techniques are the keys to improve the prognosis of patients.</p>

Diagnosis and treatment for biliary complications of hepatic cystic echinococcosis / 中华肝胆外科杂志

Objective To evaluate the diagnosis and treatment of hepatic cystic echinococcosis with biliary complications. Methods 284 patients with hepatic cystic echinococcosis (CE) with biliary complications were surgically treated from January 2002 to January 2009 in our hospital. A summary of the surgical procedures was categorized and compared in the current study. Results (1) Intrabiliary rupture of CE with obstructive jaundice and (or) inflammation of bile duct (51 patients). The diagnosis of biliary complications of hepatic hydatid cyst was difficult on ultrasound and CT, with sensitivity rates of 78.4% and 85.7%, respectively. MRCP was an effective, noninvasive and useful diagnostic tool in difficult cases; ERCP was used as the gold standard in confirmation. Biliary fistulae were seen in 3 patients (10.7%) treated by suturing the rupture site. In the non-sutured group, 17 patients (74%) developed biliary fistulae after surgery (P＜0.01). In three patients the fistula was a high-output type (the fistula output was greater than 250 ml/d). (2) CE communicated with the bile duct and (or) infection (210 patients): The cavity-related problems and draining time in group C (no bile duct exploration and decompression) were significantly higher than group A (biliary system explored and decompressed through the cystic duct) and group B (biliary system explored and decompressed through the common bile duct), while cavity-related problems and draining time between the A and B groups showed no significant difference. Biliary tract-related problems in group A was significantly lower than group B (P＜0. 05). Conclusions (1) MRCP was an effective, noninvasive and useful diagnostic tool; ERCP was used only as the gold standard in confirming intrabiliary rupture of liver cystic hydatid disease, and also as an effective technique for treating extended postoperative external biliary fistula. (2) This study indicated that suturing the communication at the rupture site and biliary decompression were effective with low morbidity and mortality rates. (3) Cholangiography and common bile duct exploration through the cystic duct could solve the cavity-related problems while avoiding the T-tube related problems.

Stochastic clinical contrastive study on two reconstructive methods after resection of cardiac carcino- ma / 肿瘤研究与临床

Objective To explore the reconstructive methods of keeping cardiac function after resec- tion of cardiac carcinoma. Methods After anastomosis of esophageal mucosa and sub-mueosa with gastric counterparts, oblique invagination with gastric ehorion and visceral muscle were adopted in experimental group to keep cardiac anatomical and physiological function.Additionaly, tissues around pylorus were suffi- ciently dissociated to keep pylorie function.Contrastively,regular anastomosis with GF-1 26/28 after resection of cardiac carcinoma was adopted to reconstruct cardia,with thread marked 4 intervally used to reinforce anastomotic ostium in control group.The sphincter of pylorus was dilated to prevent pylorospasm and obstruc- tion.Results In experimental group,no anastomotic leakage,anastomotic stricture and gastroesophageal re- flux occurred.In control group,the incidence of anastomotic leakage and anastomotic stricture were 2.8 %(1/ 36)and 50 %(18/36)respectively,and pH≤4 at anastomotic ostium were found in 12 eases,accounting for 33.3 %, and pH≤6 in 29 cases, accounting for 80.6 %(29/36). Conclusion The reconstructive method of keeping cardiac and pyloric function in resection of cardiac carcinoma benefits recovery of post-operation pa- tients and improve their life quality.

Antisense oligonucleotide inhibition of coxsackievirus B3 gene expression in HeLa cells and dose-response experiments / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In this study, the authors investigated inhibition of coxsackievirus B (CVB) gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon and structural protein coding sequences and also observed the dose-response of the sequence specific inhibition of CVB plaque formation by antisense oligonucleotides.</p><p><b>METHODS</b>Antiviral activities of these oligonucleotides were evaluated by using plaque reduction assay, yield reduction assay, cytopathic effect (CPE) and Western blot analysis. The cells were treated with random oligonucleotides as a specificity control.</p><p><b>RESULTS</b>At a screening concentration of 5 micromole, 6 of the phosphorothioate oligonucleotide demonstrated some reduction of virus replication relative to untreated cells. 70%-90% inhibition of virus at 0.1 MOI (multiplicity of infection), 50% inhibition of virus infection at 10 MOI. The levels of the VP1 were reduced in CVB-infected cells treated with Scb561 and Scb733. VP1 was significantly reduced after treatment with 0.625 micromole Scb561 and almost undetectable in cells treated with 2.5 micromole Scb561. Dose response experiments implied that sequence specific oligonucleotide doses were related to effect on inhibition of CVB3 infection. When oligonucleotide doses were increased from 1.25 to 5 micromole, 75% to 90% inhibition were observed with Scb561 and 65% to 80% inhibition with Scb733, whereas random control failed to inhibit CVB replication (8% inhibition for each). CONCLUSION The present studies showed that antisense oligonucleotides against internal ribosome entry site (IRES) and translation initiation codon were capable of specifically inhibiting the synthesis of viral protein and subsequent productive CVB replication.The selective inhibition using antisense oligonucleotide might lead to development of an effective antiviral agent for future clinical evaluation.</p>

Effects of microcystins on cell cycle and expressions of c-fos and c-jun / 中华预防医学杂志

<p><b>OBJECTIVES</b>To investigate the effects of microcystins on cell cycle and expressions of c-fos and c-jun, and explore the potential carcinogenic mechanisms of Microcystins.</p><p><b>METHODS</b>Microcystic cyanobacteria extraction (MCE) purified by Sep-Pak C(18) cartridge was added into the media and co-incubated with SHE cell for various periods. Immunohistochemistry assay was applied to detect the expressions of c-fos and c-jun at 1, 3, 6 hr time point, and cell cycle at 6, 12, 24 hr point were analyzed by flow cytometry respectively.</p><p><b>RESULTS</b>Sustained up-regulated expression of c-fos and c-jun were induced by MCE during the experimental period, and 5 - 6 folds increased expression were observed at 6 hour point after treatment. As much as 44.8 per cent of cells were induced to entry S-phase from resting G(0)/G(1).</p><p><b>CONCLUSION</b>Up-regulating the expression of transcript factor such as c-fos and c-jun thus to induce cell abnormal proliferation may be the potential carcinogenic mechanisms of microcystins.</p>

Progress on Carcinogenicity of Alga Toxins / 环境与健康杂志

Three aspects of advances in study on the carcinogenicity of alga toxins are reviewed in this paper. Water pollution by alga toxins has become a worldwide environmental problem, and special attention has been paid to the potential carcinogenicity of microcystins, the main alga toxin. Results from the study on structure_activity relationship, short_term screening assay in vitro and long_term rodent carcinogenesis bioassay all showed that alga toxins possessed tumor_promoting activities. It suggested that inhibition of protein phosphatase activities, induction of expression changes of cell proliferation_related genes, dysregulations of signal transduction and cell proliferation might be the mechanisms of the carcinogenesis effect of alga toxins.