Relation among p130Cas, E-cadherin and beta-catenin expression, clinicopathologic significance and prognosis in human hepatocellular carcinoma.

BACKGROUND: p130Cas (p130Crk-associated substance) is a junction protein that is important to the adhesion between cytoskeleton and extracellular matrix. Also, the adhesion molecules E-cadherin and beta-catenin play important roles in the invasiveness of carcinoma. This study was undertaken to investigate the effects of p130Cas, E-cadherin and beta-catenin on the invasion, metastasis and prognosis of hepatocellular carcinoma (HCC). METHODS: Immunohistochemistry was used to evaluate the expression of p130Cas, E-cadherin, and beta-catenin in 40 patients with HCC. All patients were followed up postoperatively, and the relationship between expression and clinicopathological prognostic parameters was analyzed. RESULTS: The positive expression rates of p130Cas and E-cadherin in HCC tissue (n=40) were 62.50% and 55.00%, but in normal liver tissue 10%, and 100%, respectively (P<0.05). The abnormal expression rate of beta-catenin in HCC tissue was 70%, while in normal liver tissue it was 13.33% (P<0.05). The positive rate of p130Cas was correlated with lymph node invasion, pathological stage, TNM stage, and a worse prognosis, but not with gender, age, HBV infection, hepatic cirrhosis, alpha-fetoprotein (AFP) level before operation, and tumor diameter. Similarly, the expression of E-cadherin and beta-catenin was correlated with lymph node invasion, pathological stage, TNM stage, and worse prognosis, but not with gender, age, HBV infection, hepatic cirrhosis, AFP level before operation, and tumor size. Correlations were found between p130Cas and abnormal E-cadherin/beta-catenin expression (P<0.001 and <0.05, respectively). CONCLUSIONS: In HCC, there is a negative correlation between the positive expression of p130Cas and the normal expression of the adhesion molecules E-cadherin/beta-catenin, and p130Cas plays important roles in the invasion, metastasis and prognosis of HCC. p130Cas may be involved in alterating the structure and function of E-cadherin/beta-catenin, by regulating tyrosine phosphorylation via the p130Cas-Src signal pathway.

Mechanism of benign biliary stricture: a morphological and immunohistochemical study.

AIM: To explore the mechanism of benign biliary stricture. METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on day 3, in wk 1, 3 and mo 3, 6 after operation. CD68, TGF-beta1 and alpha-SMA were examined by immunohistochemical staining, respectively. RESULTS: The mucosal epithelium of the bile duct was slowly recovered, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix was over-deposited. Myofibroblasts functioned actively and lasted through the whole process. The expression of macrophages in lamina propria under mucosa, TGF-beta1 in granulation tissue, fibroblasts and endothelial cells of blood vessels, alpha-SMA in myofibroblasts were rather strong from the 1st wk to the 6th mo after operation. CONCLUSION: The type of healing occurring in bile duct belongs to overhealing. Myofibroblasts are the main cause for scar contracture and stricture of bile duct. High expressions of CD68, TGF-beta1 and alpha-SMA are closely related to the active proliferation of fibroblasts, extracellular matrix over-deposition and scar contracture of bile duct.

Clinical Analysis of Intraperitoneal Lymphangioma.

BACKGROUND: Intraperitoneal lymphangioma (IL) used to be thought of as a benign lymphatic malformation with a low rate of preoperative diagnosis. This retrospective study aimed to explore the connection between the cysts and clinical manifestation and imaging characteristics, and to study diagnostic confusion, therapeutic principles and potential recurrent reasons, to further enhance the comprehension of this rare disease. METHODS: Here, we retrospectively reviewed 21 patients diagnosed with IL. Age, sex, complaints, physical findings, and imaging features of each patient were documented. The therapies, postoperative complications and treatments were discussed. RESULTS: Symptomatology included eight patients (38%) with intermittent dull pain in the abdomen, and three patients (14%) complained of abdominal persistent pain. The physical examination revealed an abdominal mass in 16 patients (76%), and eight (38%) were reported no discomfort. IL was correctly established preoperatively in 19 patients (90%). Patients were treated using laparotomy, except one who was treated with laparoscopy. Two recurrences were noted during follow-up. CONCLUSIONS: IL should be suspected in any patient with a mobile abdominal mass and surgery is required immediately after discovery of the tumor.

Systematic review and meta-analysis of prophylactic abdominal drainage after pancreatic resection.

AIM: To investigate whether prophylactic abdominal drainage is necessary after pancreatic resection. METHODS: PubMed, Web of Science, and the Cochrane Library were systematically searched to obtain relevant articles published before January 2014. Publications were retrieved if they met the selection criteria. The outcomes of interest included: mortality, morbidity, postoperative pancreatic fistula (POPF), clinically relevant pancreatic fistula (CR-PF), abdominal abscess, reoperation rate, the rate of interventional radiology drainage, and the length of hospital stay. Subgroup analyses were also performed for pancreaticoduodenectomy (PD) and for distal pancreatectomy. Begg's funnel plot and the Egger regression test were employed to assess potential publication bias. RESULTS: Nine eligible studies involving a total of 2794 patients were identified and included in this meta-analysis. Of the included patients, 1373 received prophylactic abdominal drainage. A fixed-effects model meta-analysis showed that placement of prophylactic drainage did not have beneficial effects on clinical outcomes, including morbidity, POPF, CR-PF, reoperation, interventional radiology drainage, and length of hospital stay (Ps > 0.05). In addition, prophylactic drainage did not significantly increase the risk of abdominal abscess. Overall analysis showed that omitting prophylactic abdominal drainage resulted in higher mortality after pancreatectomy (OR = 1.56; 95%CI: 0.93-2.92). Subgroup analysis of PD showed similar results to those in the overall analysis. Elimination of prophylactic abdominal drainage after PD led to a significant increase in mortality (OR = 2.39; 95%CI: 1.22-4.69; P = 0.01). CONCLUSION: Prophylactic abdominal drainage after pancreatic resection is still necessary, though more evidence from randomized controlled trials assessing prophylactic drainage after PD and distal pancreatectomy are needed.

Dexamethasone and dextran 40 treatment of 32 patients with severe acute pancreatitis.

AIM: Based on the pathogenesis of severe acute pancreatitis and our experimental studies, to investigate the effect of dexamethasone and dextran in treatment of patients with severe acute pancreatitis. METHODS: Thirty-two patients with severe acute pancreatitis were treated with 0.5-1 mg/kg per day dexamethasone for 3-5 d, and 500-1,000 mL/d of dextran 40 for 7 d, besides the routine therapy. RESULTS: After 4-8 h of treatment, abdominal pain began to be relieved; range of tenderness began to be localized in 27 patients. They were cured with nonsurgical treatment. Five of them were deteriorated, and treated with surgery. Four patients in this group died. CONCLUSION: Dexamethasone and dextran 40 block the pathologic process of severe acute pancreatitis through inhibition of inflammatory mediators and improvement of microcirculation disorders respectively.

Effect of spleen on immune function of rats with liver cancer complicated by liver cirrhosis.

OBJECTIVE: To establish a rat model of liver cancer complicated by liver cirrhosis and explore the effects of the spleen on immune function in this model. METHODS: Liver cirrhosis was inflicted in rats by percutaneous injection of 40% CCl4 on the back. Walker-256 tumor cells were inoculated in the cirrhotic liver and splenectomy was performed. Two weeks later, the growth and metastasis of tumor were observed and the amount of ascites and the activity of NK cells and CD25 cells were investigated. RESULTS: The amount of ascites and tumor volume were significantly higher in splenectomy group than in controls (P<0.01). Two weeks after inoculation, the activity of NK cells in both groups was decreased as compared with that before the inoculation (P<0.01). There was no significant difference between the two groups before and after the inoculation (P>0.05). The number of CD25 in both groups was higher than that before the inoculation (P<0.01). However, there was no significant difference between the two groups before and after the inoculation (P>0.05). CONCLUSIONS: Splenectomy in early stage of tumor inoculation can stimulate the tumor growth and metastasis. The activity of NK cells and the number of CD25 are inhibited by tumor itself, not by splenectomy.

Reoperation for benign biliary tract diseases in 149 cases: causes and prevention.

BACKGROUND: Failure to diagnose and treat benign biliary tract disease relatively common surgical disease may cause serious consequences. Since the introduction of B-mode ultrasonography, CT, or MRI early and accurate diagnosis of the disease has been possible. In clinical practice, however, these methods have not been adequately used. Inappropriate surgical procedures can also lead to bile duct injury or stenosis after injury, residual cholecystitis, stenosis after cholangiojejunostomy, or stenosis of the Oddi's sphincter. But improvement of the diagnosis and treatment of benign biliary tract disease remains a great challenge to clinicians. METHODS: A total of 149 patients with benign biliary tract disease who had received reoperation from June 1988 to June 2001 were analyzed retrospectively. Among them 95 patients (63.76%) received operation twice and 38 (25.5%) underwent 3 operations. Sixteen patients (10.74%) needed 4 or more operations. The procedures for the first operation included cholecystectomy (71 patients, 47.65%), cholecystectomy with exploration of the common bile duct (42, 28.19%), cholangiojejunostomy (21, 14.1%), and laparoscopic cholecystectomy (15, 10.06%). RESULTS: The causes for reoperation included residual and recurrent bile duct stones in 53 patients (35.57%), bile duct injury or stenosis after injury in 41 (27.52%), residual cholecystitis with or without stones in 28 (18.8%), stenosis after cholangiojejunostomy in 17 (11.41%), stenosis of the Oddi's sphincter in 5 (5.35%), and others in 5 (5.35%). Four patients (2.68%) died after operation. CONCLUSIONS: To prevent reoperation for benign biliary tract diseases, the following measures should be taken to increase preoperative diagnostic rate, to understand conditions of the biliary tract by using imaging techniques and cholangiography, to examine comprehensively and carefully with choledochoscopy, cholangiography and B-mode ultrasonography intraoperatively, to choose appropriate operative procedures to decrease the rate of residual stones, and to decide the time for the first repair according to injury type of the bile duct. Roux-en-Y hepaticojejunostomy with cholangioplasty is the best operation for the reconstruction of the biliary tract.

[Extraintestinal dissemination of rotavirus in immunodeficient mice].

OBJECTIVE: To investigate the extraintestinal dissemination of rotavirus (RV) in immunodeficient mice. METHODS: Immunodeficiency mouse model was established by injection of cyclophosphamide into the abdominal cavities of normal mice, then to which RV was administered either orally or intra-abdominally. The pathological changes in the organs were observed by light microscopy and RV was detected by in situ hybridization and PCR. RESULTS: Small intestinal villi, gastric lamina propria and cardiac myocytes exhibited pathological changes in the mice with oral RV administration. Besides these changes, the mice with intra-abdominal RV injection showed changes in the liver and kidneys. The intestinal villi of the mice with oral RV were RV positive by in situ hybridization. Positive results of RV in in situ PCR detection were found in the intestinal villi, intestinal gland cells, epithelial cells of the proximal convoluted tubules and collecting tubes in the kidney of the mice taken RV orally, and in the intestinal villi, kidneys, liver, heart and pancrease of mice with intra-abdominal RV injection. CONCLUSION: Immunodeficiency may be the important factor for inducing and aggravating the infection and extraintestinal dissemination of RV.

[Preliminary study of sodium butyrate-induced expression of lung resistance-related protein in K562 cells].

OBJECTIVE: To investigate the effect of sodium butyrate (NaB) on the expression level of lung resistance-related protein (LRP) in K562 cells. METHODS: K562 cells, used as an in vitro model system, were treated with NaB to induce differentiation of the cells. LRP mRNA expression and protein levels in the cells were detected by reverse transcriptase-PCR (RT-PCR) and flow cytometry, respectively. RESULTS: A 1-day treatment with 2 mmol/L NaB induced the gene expression of LRP to increase to the maximum in the cells, and a 3-day treatment caused the corresponding protein level to increase to the maximum. CONCLUSION: Both the mRNA level and the protein expression of LRP can be induced by NaB in K562 cells.

[Preliminary study of human calicivirus infection in Guangzhou].

OBJECTIVE: To investigate of human calicivirus (HuCV) infection in the children of Guangzhou, the capital of Guangdong Province at China, and conduct preliminary study of the prevalence of the virus. METHODS: Serum specimens from 266 children of different ages were tested by an indirect enzyme-linked immunosorbent assay (ELISA) for specific IgG against recombinant Norwalk virus particles (rNV), recombinant Mexico virus partides(rMX) and recombinat 387 Virus particles(r387), respectively, and fecal specimens were collected for detection of HuCV RNA using reverse transcriptional (RT)-PCR assisted by sequence analysis on the basis of comparison of the amplified product with the sequence entries in GenBank. RESULTS: The seroprevalence was 68.0% for rNV, 51.5% for rMX and 71.1% for r387 in the cohort of children enrolled in this study. Norwalk-like viruses (NLVs) were detected in 4 out of the 100 fecal specimens collected from cases of nonbacterial gastroenteritis during the peak season of diarrhea, and also in 3 of the 10 fecal specimens collected during the outbreak of diarrhea in an elementary school. CONCLUSIONS: Sporadic and fulminant NLV infections are present in Guangzhou. The children have the infection in their infancy, and the infection rate increases sharply with age.

[Comparison of several silver staining methods for DNA polyacrylamide gel electrophoresis].

OBJECTIVE: To compare 4 silver staining methods for DNA detection in polyacrylamide gel electrophoresis. METHODS: After the electrophoresis was completed, the gels were stained separately by four different methods, namely using ammonia-silver-citric acid, low-concentration silver nitrate, 0.1% and 0.2% silver nitrate. RESULTS: DNA was detected only by staining with 0.1% and 0.2% silver nitrate. CONCLUSION: 0.2% silver nitrate used along with sodium hydroxide shows the highest sensitivity in DNA detection, while its use with sodium carbonate produces the best quality of the image.

Induction of erythroid differentiadon in K562 cells by different butyrate regimens.

OBJECTIVE: To investigate the hemoglobinization induced by butyrate and observe the effects of different butyrate regimens on erythroid differentiation of K562 cells. METHODS: K562 cells, used as an in vitro model system, were stained with benzidine to assess hemoglobin (Hb) production in response to different treatment regimens of butyrate at varied concentrations. Comparison of the percentage of benzidine-positive cells (BZ%)in untreated and butyrate-treated K562 cells was performed. Protein absorption at 414 nm using a spectrophotometer and cellulose acetate gel electrophoresis were employed to determine the changes of Hb production in K562 cells. RESULT: The BZ% increased by 4 to 6 fold and Hb production by 9 to 14 fold 3 d after the cells were incubated with butyrate which selectively promoted fetal hemoglobin(HbF) production in K562 cells. The BZ% increased gradually and reached the peak of l9% to 28% on day 3 or 4 in cells receiving pulse treatment with butyrate for only once, followed by a subsequent rapid fall and on day 7 to 9, it decreased to the level of untreated K562 cells. The length of time for incubation with butyrate was not related to in the increment or the maintenance of the increased level of BZ%. Continuous treatment with butyrate yielded a similar result to that of a single administration of pulse treatment. In contrast, in cells with intermittent pulse treatment the BZ% reached a peak after 72 h and was maintained between 20% and 30% till 3 cycles of treatment was completed. CONCLUSION: Butyrate can induce the expression of globin genes and augment Hb producfion especially that of HbF. A sustained erythroid differentiation of K562 cells can be achieved by intermittent pulse treatment with butyrate which can be an ideal regimen for children with beta globin diseases.

MACC1 expression correlates with PFKFB2 and survival in hepatocellular carcinoma.

OBJECTIVE: To validate the relationship between MACC1 and 6-phosphofructo-2-kinase/fructose 2, 6 bisphosphatase (PFKFB2) expression as well as its clinicopathological features and prognostic significance in hepatocellular carcinoma. METHODS: By using immunohistochemistry, we investigated the MACC1 and PFKFB2 protein expression in 60 pairs of hepatocellular carcinoma and corresponding non-tumor tissues. Using the Mann-Whitney U test, the Chi-square test, Kaplan-Meier survival analysis, Cox proportional hazard regression analysis and Spearman analysis, we studied the relationship between MACC1 and PFKFB2 protein expression and postoperative overall survival (OS) of the HCC patients. RESULTS: MACC1 and PFKFB2 positive staining rates were significantly higher in hepatocellular carcinoma than in the corresponding nontumor tissues (P=0.012 and 0.04, respectively). The clinicopathological features evaluation revealed that positive expression of MACC1 was associated with a high Edmondson classification (P=0.007) and advanced TNM stage (P=0.027). Similar findings were evident for PFKFB2 expression (P=0.002 and P=0.027). MACC1 and PFKFB2 positive expression was associated with a lower OS rate (P=0.004 and 0.03, respectively). Kaplan-Meier survival and Cox proportional hazard regression analyses revealed MACC1 positive expression to be a prognostic factor for postoperative OS, but PFKFB was not. CONCLUSION: Highly expressed MACC1 and PFKFB2 protein were associated with TNM stage, Edmondson -Steier classification and overall survival. MACC1 may affect tumor metabolism partly through expression and phophorylation of PFKFB2.

[Neonatal hyperthyroidism: a case report and literature review].

OBJECTIVE: We report a case of neonatal thyrotoxicosis with concurrent respiratory failure in an infant born to a mother with Graves' disease and review the published literature describing neonatal hyperthyroidism. The male infant who was born by spontaneous delivery at 35 weeks of gestational age presented with fever, tachycardia and tachypnea at rest on day 11 after birth, and developed severe apnea on day 14. Thyroid function studies revealed hyperthyroidism in the infant, and his mother was confirmed to have Grave's disease during pregnancy. Literature review showed that among the 33 infants with similar conditions, tachycardia, tachypnea and poor weight gain were the most distinct clinical features of congenital hyperthyroidism. Accurate diagnosis of Graves' disease in the mother during pregnancy and awareness of the clinical presentations of neonatal hyperthyroidism are key to reducing missed diagnosis or misdiagnosis of neonatal hyperthyroidism.

[Expression of Fbxw7 and its correlation with cell proliferation in human hepatocellular carcinoma].

AIM: To observe the expression of Fbxw7 and explore its correlation with cell proliferation of hepatocellular carcinoma (HCC). METHODS: The expression of Fbxw7 in paired tumor and tumor-adjacent normal tissues from 40 patients with HCC were assessed using RT-PCR and immunohistochemistry. The expression level of Fbxw7 mRNA in normal liver cell line (LO2) and HCC cell lines (Hep3B and SMMC-7721) were detected by real-time RT-PCR. Colony formation assay and tumor xenograft assay were performed in different cell lines with different Fbxw7 expression respectively to investigate the ability of cell proliferation in vitro and in vivo. RESULTS: The expression of Fbxw7 at both mRNA and protein levels in HCC tissues were significantly down-regulated compared to tumor-adjacent normal tissues (P<0.05). Down-regulation of Fbxw7 was found apparently associated with high Edmonson-Steiner classification and advanced TNM stage (P<0.05). The expression of Fbxw7 mRNA in LO2 was significantly higher than that in Hep3B or SMMC-7721 (P<0.05). Cell lines with less Fbxw7 expression had not only more colonies in colony formation assay (P<0.05), but also bigger xenografts in vitro (P<0.05). CONCLUSION: The low expression of Fbxw7 correlates with the poor clinicopathological characteristics and cell proliferation of HCC.

[Gli1 expression and its relationship with the expression of Shh, Vimentin and E-cadherin in human hepatocellular carcinoma].

AIM: To investigate the expression and clinical features of glima-associated oncogene 1(Gli1) and its correlation with the expression of sonic hedgehog(Shh), one of the ligands of hedgehog (Hh) signaling, and two epithelial-mesenchymal transition (EMT) markers, Vimentin and E-cadherin in human hepatocellular carcinoma(HCC). METHODS: Paired HCC and normal tumor-adjacent tissues were collected from 63 HCC patients. Gli1 expression at both the protein and mRNA level were examined by immunohistochemistry and RT-PCR. The protein expression of Shh, Vimentin and E-cadherin were evaluated by immunohistochemistry to identify correlations with Gli1. RESULTS: The protein and mRNA expression of Gli1 were significantly up-regulated in the HCC tumor tissues compared to the normal tumor-adjacent tissues. Gli1 protein expression in HCC was closely correlated with intrahepatic metastases (x(2);=6.205, P<0.05), portal vein invasion (x(2);=4.014, P<0.05), high Edmonson-Steiner classification (x(2);=19.668, P<0.05) and advanced TNM stage (x(2);=7.091, P<0.05). Gli1 protein expression was positively correlated with Shh (r=0.574, P<0.05) and Vimentin(r=0.467, P<0.05), and negatively correlated with E-cadherin (r=-0.439, P<0.05). CONCLUSION: Gli1 is up-regulated in HCC tissues and closely correlated with clinicopathological characteristics, the increased expression of Gli1 in HCC tissues may be attributed to Shh, and Gli1 may play an important role in HCC progression and metastasis by inducing EMT.

[Downregulation of HSP70 gene expression and apoptosis in human hepatocellular carcinoma SMMC-7721 cells induced by nimesulide in vitro].

AIM: To investigate the effect of nimesulide on cell apoptosis and possible mechanism in human hepatocellular carcinoma SMMC-7721 cells. METHODS: SMMC-7721 cells were treated with nimesulide at different concentrations. Cell viability was assessed by MTT assay. Cell apoptosis rate was determined with flow cytometry. The cleavage activity of PARP and caspase-9 and the expression of HSP70 were evaluated using RT-PCR and Western blotting. The influence of HSP70 on cell apoptosis was observed using RNA interference silencing HSP70 expression. RESULTS: Nimesulide significantly inhibited cell growth in SMMC-7721 cells in a time- and concentration-dependent manner, and induced cell apoptosis in a concentration-dependent manner. Moreover, nimesulide promoted the cleavage of caspase-9 and PARP and inhibited the mRNA and protein expression of HSP70. Through the specific inhibition on HSP70 gene with siRNA, cell apoptosis increased, and the apoptosis was enhanced by the cleavage activity of caspase-9 and PARP. CONCLUSION: Nimesulide could inhibit cell growth and induce apoptosis in human hepatocellular carcinoma SMMC-7721 cells via the downregulation of HSP70.

[UbcH10 expression in hepatocellular carcinoma and its clinicopathological significance].

OBJECTIVE: To investigate UbcH10 expression in hepatocellular carcinoma and explore its clinicopathological implications. METHODS: We detected UbcH10 mRNA expression using RT-PCR in normal liver cell line, cancer cell lines, surgically removed hepatocellular carcinoma tissue and corresponding adjacent non-tumor tissue and evaluated the clinicopathological significance of UbcH10. Immunohistochemistry was performed to investigate UbcH10 protein expression in hepatocellular carcinoma tissue, the adjacent tissue, and normal liver tissue specimens. RESULTS: Normal liver cell line L02 showed significantly lower UbcH10 mRNA expression levels than the cancer cell lines BEL-7402, Hep3B, HepG2 and SMMC-7721 (P<0.05). UbcH10 mRNA expression was also was significantly higher in hepatocellular carcinoma tissues than in the corresponding non-tumor tissues (P<0.05). Clinicopathological evaluation suggested that UbcH10 expression was associated with tumor invasion of the portal vein, tumor size, TNM staging, and tumor differentiation (P<0.05). Immunohistochemistry identified stronger UbcH10 expression in hepatocellular carcinoma tissues than in the adjacent tissues and normal liver tissues (68.6%, 28.6%, and 26.7%, respectively). CONCLUSION: UbcH10 is over-expressed in hepatocellular carcinoma and may serve as a novel biomarker as well as a therapeutic target of hepatocellular carcinoma.

[Alveolar capillary dysplasia: a case report and review of literature].

OBJECTIVE: To report a newborn infant who died of alveolar capillary dysplasia (ACD). The literature on about 20 cases of ACD was reviewed. METHODS: A retrospective review of records of infants from Medline with a diagnosis of ACD was carried out. RESULTS: The case was a newborn female infant who developed respiratory distress 5 hours after an uncomplicated delivery. She died at the fourth day after birth despite full ventilatory support. The lung autopsy provided a diagnosis of ACD. In the 21 infants, 7 were male and 14 were female; 19 infants were born full-term and 2 were born pre-term. The birth weight of 19 infants and Apgar score of 15 infants were normal; 16 infants developed progressing tachypnea and cyanosis within 24 hours of age, 5 developed cyanosis at 1 day to 19 days. Echocardiography demonstrated a right to left shunt in the hearts of all the 21 infants, and pulmonary hypertension in 20 infants. Twenty infants were treated with conventional mechanical ventilation, 7 infants with high-frequency oscillatory ventilation and 12 infants with extracorporeal membrane oxygenation (ECMO). Fourteen infants were also treated with inhaled nitric oxide therapy and 4 with exogenous surfactant. Diagnostic open lung biopsy was performed in 6 infants. The chest radiography showed normal findings in 3 infants, pneumothoraces in 9 infants, reticular markings, granular, patchy or diffuse opacity in lungs of 7 infants, and decreased pulmonary vascular markings in two infants. All the 21 infants died; 8 of them died within 10 days of age, 7 within 30 days of age, and one died at the age of 4 months who was the longest survivor. Fourteen infants were associated with congenital malformations, such as cardiovascular, gastrointestinal, and genitourinary systems, including one infant associated with chromosomal abnormalities, two infants of familial genetic predisposition. CONCLUSIONS: At present, ACD is still a disease with poor prognosis, significant medical expenses and no specific treatment. When respiratory failure or persistent pulmonary hypertension (PPHN) is persistent after routine treatment in an infant, ACD should be highly suspected and conventional open-lung biopsy should be preformed to confirm the diagnosis.

[Application of a tumor cell vaccine transfected with GM-CSF/IL-2 fusion gene for specific immunotherapy of hepatocellular carcinoma].

OBJECTIVE: To prepare a transgenic tumor cell vaccine transfected the fusion gene of murine granulocyte-monocyte colony stimulating factor (mGM-CSF) and human interleukin-2 (hIL-2) using H22 cells, and explore its specific antitumor immunity against hepatocellular carcinoma. METHODS: The eukaryotic vector expressing the fusion gene mGM-CSF/hIL-2 was transfected into H22 cells followed by radiation exposure to construct the tumor cell vaccine, which was used to immunize Balb/c mice by subcutaneous inoculation. The mice inoculated subcutaneously with H22 cells, cells transfected with the empty vector pcDNA(+), or with PBS served as the controls. A week later, H22 cells were injected peritoneally into Balb/c mice for establishing the tumor-bearing model, and their serum levels of interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) were detected using enzyme-linked immunosorbent assay (ELISA) with the survival of the mice recorded. The spleen cells were obtained from the mice immunized with the tumor cell vaccine, the tumor-bearing mice and the normal control mice to assess their cytotoxicity against the parental H22 cells in vitro using (51)C(r)-release assay. RESULTS: The transgenic H22 cell vaccine transfected with mGM-CSF/hIL-2 fusion gene was successfully constructed. The killing rate of H22 cells by the spleen cells from the mice immunized with the transgenic cell vaccine was significantly higher than those by the spleen cells from the tumor-bearing mice or normal control mice (38.3% vs 13.6% and 7.5%, P<0.05). Serum IFN-gamma in the tumor-bearing mice immunized with the transgenic cell vaccine was significantly higher, and serum IL-10 significantly lower than those of the control groups (P<0.01). The survival time of the tumor-bearing mice injected with the transgenic cell vaccine was also significantly prolonged. CONCLUSION: Syngeneic tumor cell vaccine genetically modified by mGM-CSF/hIL-2 fusion gene transfection can elicit specific cellular immune response and enhance the host antitumor immune response to extend the survival time of tumor-bearing mice.