Expression and significance of prostate stem cell antigen and Oct-4 in benign and malignant lesions of the stomach / 中南大学学报(医学版)

OBJECTIVE@#To determine on the expression of prostate stem cell antigen (PCSA )and Oct-4 and detect their clinicopathological significance in benign and malignant lesions of the stomach.@*METHODS@#EnVision immunohistochemistry for assaying the expression of PSCA and Oct-4 was used in paraffin-embedded sections from specimens of primary foci (n = 58) and metastatic foci of regional lymph nodes (n = 36) of gastric cancer, peritumoral tissues (n = 20), and benign lesions of the stomach (n = 80).@*RESULTS@#The positive rates of PSCA and Oct-4 were significantly higher in gastric cancer than those in peritumoral tissues and benign lesions (P 0.05). The positive rates of PSCA and Oct-4 were significantly lower in infiltrating depth T(1) to approximately T(2), non-metastasis of lymph nodes, metastasis of lymph nodes N1 site, and non-metastasis of distant organs than those in infiltrating depth T(3) to approximately T(4), metastasis of lymph nodes, metastasis of lymph node N(2) to approximately N(3) site, and metastasis of distant organs(P < 0.05 or P < 0.01).Conclusion The expressive levels of PSCA and Oct-4 might be related to the invasive potential, metastasis of lymph nodes, and distant organs, it suggested the expressions levels of PSCA and Oct-4 might be important markers for reflecting the biological behaviors and prognosis of gastric cancer.

Screening up-regulated genes in hepatic stellate cells treated with PDGF-BB using suppression subtractive hybridization technique / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct a subtractive cDNAs library of up-regulated genes in rat hepatic stellate cells (HSCs) stimulated with platelet-derived growth factor (PDGF)-BB by suppression subtractive hybridization (SSH) technique, to clone the up-regulated genes associated with its regulation effects, and to elucidate the mechanism of the molecular biology of hepatic fibrosis involved in PDGF-BB.</p><p><b>METHODS</b>The mRNA was isolated from HSCs stimulated with PDGF-BB and controlled with identical cells untreated with PDGF-BB, and then the cDNAs were synthesized. The cDNAs were designated as tester and driver. After being digested by restriction enzyme Rsa I, small-sized cDNAs were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptor 1 and adaptor 2, individually. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, the amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5alpha. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR.</p><p><b>RESULTS</b>The subtractive cDNAs library of up-regulated genes in HSCs stimulated with PDGF-BB was constructed successfully. The amplified library contained 102 positive clones. Colony PCR showed that 93 clones contained 200-1000 bp inserts. Sequence analysis was performed in 31 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank; altogether 13 coding sequences were obtained, which were known ones. The genes mainly included voltage-dependent anion channel (VDAC), heat shock protein 47 and RAN-member RAS oncogene family genes.</p><p><b>CONCLUSION</b>Acting as one of the most effective mitogens, PDGF-BB up-regulated some gene expressions during stimulation of the HSCs, including some cell growth associated proteins, some proteins participating in intracellular metabolism and some molecular chaperone proteins. This work brings some new clues for studying the molecular biological mechanism involved in the up-regulated genes in PDGF-BB transactivated HSCs in hepatic fibrosis.</p>

A study on the anti-HBV effect of dendritic cell from human umbilical cord blood / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Regarding the strong antigen-presenting abilities of dendritic cells (DC), this study was carried out based on the induction and proliferation of DC derived from human umbilical cord blood; the anti-HBV effect of cytotoxicity T lymphocytes (CTL) activated by those DC pulsed with HBsAg was also carried out to explore a new way to activate the HBsAg-specific CTL.</p><p><b>METHODS</b>Cord blood was collected from the cord veins of normal placentae after Cesarean sections, from which cord blood mononuclear cells (CBMC) were separated through density gradient centrifugation. The CBMC were cultured in RPMI 1640 with a cytokine cocktail. Pulsed with HBsAg, the DC were prepared to activate the HBsAg-specific CTL among the CBMC. The cytotoxic effect of CBMCs activated by the DC primed with HBsAg was assayed through the killing of those HepG2-S target cells.</p><p><b>RESULTS</b>Typical DC could be induced from CMBC cultured with a cytokine cocktail. DC pulsed by HBsAg activated HBsAg-specific CTL, which killed the target HepG2-S cells to some extent.</p><p><b>CONCLUSION</b>DC can be induced from CMBC with the cytokine cocktail and they show a strong antigen-presenting ability. DC produced in this way and pulsed by HBsAg can activate HBsAg-specific CTL in vitro. This might mean that it could be a new way to break the tolerance to HBV in chronic HBV-infected patients.</p>

In situ hybridization study on the expression of Kiss-1 and KAI-1 metastasis suppressor genes in gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the mRNA levels of Kiss-1 and KAI-1 metastasis suppressor genes in gastric cancer, and to explore its clinical value.</p><p><b>METHODS</b>In situ hybridization was used on routinely paraffin-embedded sections of resected specimens of 49 cases with gastric cancer and 20 cases with pericancerous tissue.</p><p><b>RESULTS</b>The positive rates and the scores of Kiss-1 and KAI-1 mRNA in gastric cancer tissue were significantly lower than those in pericancerous tissue (P<0.01). The positive rates and scores in normal to mild-atypical hyperplasia cases were significantly higher than those in middle to severe-atypical hyperplasia of pericancerous tissue (P<0.05,P<0.01). The positive rates and scores of Kiss-1 and KAI-1 mRNA in patients with infiltrating depth T1~T2, without lymph node metastasis, with only first group of lymph node metastasis, and without distant organ metastasis were significantly higher than those in patients with infiltrating depth T3~T4, with lymph node metastasis, with second or third group of lymph node metastasis and with distant organ metastasis. A strong positive correlation was found between the expressive scores of Kiss-1 and KAI-1 mRNA in gastric cancer (r=0.53, P<0.01).</p><p><b>CONCLUSION</b>The expression of Kiss-1 and/or KAI-1 mRNA may be important biological markers of reflecting invasive and metastatic potential and prognosis in gastric cancer. The assays of Kiss-1 and/or KAI-1 mRNA expression level in benign lesions of stomach may have important clinical value for the protection and early-stage finding of gastric cancer.</p>

Effects of aminoguanidine on the lung injury induced by the total hepatic ischemia-reperfusion in rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effects of aminoguanidine on the lung injury induced by the total hepatic ischemia-reperfusion in rats.@*METHODS@#The total hepatic ischemia-reperfusion model was built after blocking of the hepatic porta, suprahepatic and infrahepatic vena cava. Ninety Sprague-Dawley rats were assigned randomly into 3 groups: Sham operation group (Group A, n=30); total hepatic ischemia group (Group B, n=30); and aminoguanidine treatment group (Group C, n=30). Each group was subdivided randomly into 3 subgroups (n=10) according to different time phases: 20 minutes after the total hepatic vascular exclusion (T0), 4 hours after the reperfusion (T1), and 48 hours after the survival Group A and Group B were intravenously injected with normal saline ( mL/kg) while Group C was injected with aminoguanidine (20 mg/kg) dissolved in normal saline (1 mL/kg) 10 minutes before the open of the abdomin. The levels of portal blood nitric oxide ( O) endotoxin ( ET), tumor necrosis factor-alpha (TNF-alpha at T0 and T1 were detected; 48 hours survival rates and the lung wet/dry weight ratio were counted; and the histological changes of the lung tissues were observed.@*RESULTS@#Compared with Group A, the levels of portal vein NO, ET, and TNF-alpha T0 and T1 in Group B and Group C were significantly higher (P < 0.05 or P < 0.01). But those indexes of Group C were lower than those of Group B (P < 0.05). The 48-hour survival rate in Group C was higher than that in Group B (P < 0.05). The lung wet/dry weight ratio in Group C was lower than in Group B (P < 0.05) and the histological change of Group C was slighter than that in Group B.@*CONCLUSION@#Aminoguanidine has the protective effects on the lungs against the total hepatic ischemia-reperfusion induced injury.

Effect of chloroquine on the apoptosis of intestinal mucosa epithelial cells and enterogenous bacteria-endotoxin translocation after total hepatic ischemia-reperfusion in rats / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of chloroquine on the apoptosis of intestinal mucosa epithelial cell and enterogenous bacteria-endotoxin translocation after total hepatic ischemia-reperfusion in rats.@*METHODS@#The rat total hepatic ischemia-reperfusion model was built by blocking the hepatic portal, suprahepatic and infrahepatic vena cava for 20 minutes. Ninety Sprague-Dawley rats were assigned randomly into the sham operation group (Group A, n = 30), total hepatic ischemia-reperfusion treatment group (Group B, n = 30), and chloroquine administrated group (Group C, n = 30). Each group was subdivided randomly into 3 subgroups (n = 10) according to different experiment time phases as follows: after 20 minutes of total hepatic vascular exclusion (T0), 4 hours after reperfusion (T1), and the 48 hours of survival. Group A and Group B were intravenously injected with normal saline 1 mL/kg while Group C received chloroquine 10 mg/kg which dissolved in 1 mL/kg normal saline intravenously. The levels of portal blood D-lactate, TNF-alpha, endotoxin, and the intestinal mucosa MDA concentration were measured at T0 and T1; the portal blood, mesenteric lymph node, and spleen tissues were cultured for bacteria; and the apoptotic index of intestinal mucosa epithelial cells at T0 and T1 and the survival rate after 48 hour reperfusion were obtained.@*RESULTS@#Compared with Group A, the levels of portal blood D-lactate, TNF-alpha, endotoxin and the intestinal mucosa MDA in Group B and Group C were significantly higher (P < 0.05 or P < 0.01). These indexes of Group C were lower than those of Group B (P < 0.05). The portal vein blood, mesenteric lymph node and spleen tissues existed the bacterium translocation both in Group B and Group C, and the positive rate in Group C was lower than that in Group B (P < 0.05). Apoptotic index of the intestinal mucosa epithelial cell increased significantly in Group B (P < 0.01) and Group C (P < 0.05), but the apoptotic index in Group C was lower than that in Group B (P < 0.05); the 48 hour survival rate of the rats in Group C was higher than that in group B (P < 0.05).@*CONCLUSION@#Chloroquine may decrease the intestinal mucosa epithelial cell apoptosis and the enterogenous bacteria-endotoxin translocation after total hepatic ischemia-reperfusion and increase the survival rate of the rats.

Effect of tumor antigen specific CTL induced by dendritic cells on a model of human hepatocellular carcinoma in nude mice (LCI-D20) / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the cure effect of tumor antigen specific CTL on a model of human hepatocellular carcinoma in nude mice LCI-D20.</p><p><b>METHODS</b>Dendritic cells (DCs) were induced from peripheral blood mononuclear cells of healthy people in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4) and were pulsed with tumor antigen from hepatocellular carcinoma cell line MHCC97H. Then tumor antigen specific cytotoxic T lymphocytes (CTLs) were induced. By intraperitoneal injection of tumour antigen specific CTLs into the LCI-D20, the preventive and therapeutic effects of these CTLs to HCC in the LCI-D20 model were assessed. Cytokine-induced killer (CIK) cells and phosphate buffer solution were used as controls at the same time.</p><p><b>RESULTS</b>The weights of tumors in the tumor antigen specific CTL group, in the CIK cell group and in the blank group were (1.11+/-0.63), (1.12+/-0.36) and (2.68+/-0.53) grams respectively (t = 5.18, t = 6.06, P < 0.01). The amount of blood alpha fetal protein in the tumor antigen specific CTL and CIK groups were (52.1+/-9.7) microg/L and (48.6+/-5.2) microg/L, and was (82.2+/-7.2) microg/L in the blank group (t = 17.26, t = 22.07, P < 0.01 respectively). The metastasis rates in livers were 16.7%, 16.7% and 58.3% in the tumor antigen specific CTL, CIK cell and blank control groups respectively (chi2= 4.44, P < 0.01). The survival time of the mice in the tumor antigen specific CTL group was (79.0+/-5.02) days, (73.3+/-7.0) days in the CIK group, and (52.3+/-5.2) days in the blank group (t = 14.56, t = 17.54, P < 0.01).</p><p><b>CONCLUSION</b>Tumor antigen specific CTLs may prevent metastasis in the LCI-D20 model and prolong the survival time.</p>

Immunotherapy with a chimeric AFP and HSP70 gene DNA vaccine targeting on a murine hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate immune responses and the anti-tumor effect of a constructed Chimeric AFP-Mt.HSP70 DNA vaccine in mice.</p><p><b>METHODS</b>Chimeric AFP-Mt.HSP70 was constructed by molecular clone techniques. Spleen cells derived from mice immunized twice were induced to secrete IFN gamma and were assayed using ELISA. The activity of the cytotoxic lymphocytes (CTL) derived from spleen cells was assayed using lactate dehydrogenase (LDH). 4 x 10(6) Hepa1-6 cells/200 microl were injected subcutaneously into the right axilla of each mouse bearing the tumor. The anti-tumor effect of the recombinant DNA vaccine was evaluated by measuring tumor sizes of the mice.</p><p><b>RESULTS</b>AFP-specific CTL reaction was induced by our chimeric DNA vaccine and Mt.HSP70 enhanced this effect (P < 0.05). The CTL activity was about 32% at E/T=50:1. The IFN gamma secreted by spleen cells of mice immunized with chimeric plasmids was about 200 pg/ml. It was higher than those in the other groups; Tumor sizes of mice immunized with fused plasmids were smaller than those in the other groups. Survival times of mice immunized with the fused plasmids were prolonged.</p><p><b>CONCLUSION</b>Chimeric DNA vaccine can induce AFP-specific CTL reaction and has an anti-tumor effect on transplanted tumors in our murine experiment.</p>

Treatment with total hepatic vascular exclusion and reperfusion for the intestinal barrier in rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the influence of treatment with total hepatic vascular exclusion and reperfusion on the intestinal barrier in rats.@*METHODS@#The total hepatic vascular exclusion and reperfusion model was built after the block of hepatic portal, suprahepatic and infraheptic vena cava for 20 minutes. Sixty Sprague-Dawley rats were divided randomly into 2 groups: sham operation group (Group A, n=30) and total hepatic vascular exclusion and reperfusion treatment group (Group B, n=30). Each group was subdivided randomly into 3 subgroups (n=10) according to different experiment time points as follows: at the end of the total hepatic vascular exclusion (T0), 4 reperfusion after total hepatic vascular exclusion (T1) and the 48 h survival. Portal vein blood gas was analysed at T0. At T0 and T1 the following items were detected: the level of portal vein blood D-lactate, tumor necrosis factor-alpha (TNF-alpha), the MDA concentration and pathologic morphology change of intestinal mucosa.@*RESULTS@#Compared with Group A, the PCO2 at T0 in Group B increased while pH, P02, and HCO3- decreased significantly (P < 0.05). The level of portal blood D-lactate, TNF-alpha and intestinal mucosa MDA at T0 and T1 was significantly higher (P < 0.05, or P < 0.01). The histologic damage in the intestinal mucosa was observed in Group B, and the rat survival in Group B was lower than that in Group A (P < 0.05).@*CONCLUSION@#The treatment with total hepatic vascular exclusion and reperfusion can damage the intestinal barrier in rats.

Enhancement of a hepatitis B DNA vaccine potency using aluminum phosphate in mice / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study antibody response to a hepatitis B DNA vaccine by formulation with aluminum phosphate in mice.</p><p><b>METHODS</b>An eukaryotic expression plasmid inserted HBsAg gene (pcDNA3.1-S) was constructed by cloning technique and the accuracy of the construct was confirmed by restriction enzyme digestion and DNA sequencing, then hepatitis B DNA vaccine formulations were prepared by mixing pcDNA3.1-S with various concentration of aluminum phosphate in 0.9% NaCl. HBsAg expressions were assayed by ELISA in vivo five days after intramuscular injection of pcDNA3.1-S with or without aluminum phosphate. And serum samples were obtained from individual immunized or control mice 6 weeks post injection. Then anti-HBs were assayed in mice sera by ELISA.</p><p><b>RESULTS</b>Five days after intramuscular immunization, the levels of HBsAg expression of groups with aluminum phosphate showed no difference from those of control group in tibialis arterials muscles. In sera, HBsAg could not be detectable in all groups. Intramuscular immunization of BABL/C mice with pcDNA3.1-S mixed aluminum phosphate (0microg, 1microg, 10microg, 50microg, 100microg) 6 weeks later, the P/N values of anti-HBs in sera were 11.54+/-5.60, 11.00+/-6.62, 20.30+/-10.20, 49.18+/-24.40 and 48.68+/-27.78, respectively. It showed that pcDNA3.1-S mixing with aluminum phosphate could increase anti-HBs titers in mice.</p><p><b>CONCLUSION</b>No increase of HBsAg expression was observed by mixing plasmid pcDNA3.1-S with various concentration of aluminum phosphate in vivo. But Intramuscular immunization of BALB/C mice with pcDNA3.1-S mixing aluminum phosphate adjuvant can increase anti -HBs titers. It seemed that aluminum phosphate would be valuable for further investigation as a potential adjuvant of hepatitis B DNA vaccines.</p>

Determination of linderane in root tuber of Lindera aggregata by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To provide scientific basis for quality control of Lindera aggregata.</p><p><b>METHOD</b>HPLC analytical method was established using a Lichrospher C18 column and acetonitrile-water (56:44) as the mobile phase, detected at 235 nm.</p><p><b>RESULT</b>The linear range of linderane is between 0.0642 - 0.5774 microg, the average recovery was 98.4%, RSD1.7% (n = 9).</p><p><b>CONCLUSION</b>Contents of linderane in commercially available and collected samples were from 0.028% to 0.123% and from 0.056% to 0.222% respectively.</p>

The detection of nanobacteria infection in serum of healthy Chinese people / 中华流行病学杂志

<p><b>OBJECTIVE</b>Nanobacteria, a new kind of bacteria found by a Finnish scholar, is considered to relate to many human diseases like nephrolithiasis. However, there are no data available on nanobacteria infection in Chinese people.</p><p><b>METHODS</b>Nanobacteria was detected in serum of 336 cases of healthy adults in Southern China by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry stain. The monoclonal antibody of nanobacterum was supplied by Kuipio University of Finland.</p><p><b>RESULTS</b>Nanobacteria infection rates were 27 (8.0%), 19 (5.7%) in the healthy adults by ELISA and immunohistochemistry stain respectively. No difference was shown between the 2 methods and between male and female, statistically. Age and sex did not seem to be related to the infectious risk of nanobacteria. However, the infectious rate was lower in those below 30-year-old than that of people over 60-year-old (P < 0.05).</p><p><b>CONCLUSION</b>Nanobacteria was existed in the serum of Chinese healthy people with an infectious rate of 8.0%.</p>

Differential gene expression of liver carcinoma cell after transfection of by ARL-1 with microarray / 中华外科杂志

<p><b>OBJECTIVE</b>To study the difference of the gene expression profile and to identify the different expression after transfection of the ARL-1 gene.</p><p><b>METHODS</b>The cDNA probes were synthesized from total RNA of study group and control group, which was differentially hybridized to cDNA chips and confirmed by a gene specific semiquantitative reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Six kinds of gene expression were increased and 9 kinds of gene expression were decreased. The findings were correlated with protein metabolism, signal pathway, metastasis, and drug resistance.</p><p><b>CONCLUSIONS</b>cDNA chips showed that gene expression profile of liver carcinoma cell was changed after transfection of the ARL-1 gene. It is a useful method in understanding the mechanism of drug resistance.</p>

The role of dendritic cell and macrophage in hepatoma antigen-presenting / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the role of dendritic cells (DCs) and macrophages, differentiated from the same individual peripheral blood monocytes, in tumor antigen- presenting.</p><p><b>METHODS</b>DCs and macrophages were differentiated from human peripheral blood monocytes by adding both Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) or GM-CSF only. Then they were loaded with tumor antigen at different concentrations and cocultured with autologous T cells in 96-well flat-bottomed microtiter plates for five days at 37 degrees C, 5% CO(2). (3)H-thymine was added before the culture terminated, and twelve hours later, the cells were gathered to test the cpm value.</p><p><b>RESULTS</b>Both DCs and macrophages chased with tumor antigen could strongly stimulate the proliferation of autologous T cells, especially DCs. The stimulation effect with 20 microl/ml antigen was the most remarkable and the cmp values were 11,950.3 +/-1621.8, 8,708.5 +/-176.1, 402.5+/-43.1 in DCs group, Macrophages group, and lymphocytes group, respectively.</p><p><b>CONCLUSION</b>The antigen presenting role of DCs is stronger than that of macrophages from the same individual.</p>

Clinical research on the treatment effect of autologous dendritic cell vaccine on the patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the treatment effect of autologous HBsAg-loaded dendritic cells (DCs) on patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>Monocytes were isolated from fresh peripheral blood of 19 CHB patients by Ficoll-Hypaque density gradient centrifugating and cultured with plastic -adherence method. DCs were induced and proliferated from the monocytes with granulocyte-macrophage clony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for seven days. After being incubated with HBsAg for two hours, DCs were injected to patients subcutaneously twice at the interval of two weeks. HBV DNA level, alanine aminotransferase (ALT) level, and HBV markers in the serum of patients were tested every two months.</p><p><b>RESULTS</b>11 of the 19 (57.9%) patients responded to DC-treatment clinically. The rates of HBeAg clearance and HBeAg/anti-HBe seroconversion were 52.6% (10/19) and 26.3% (5/19) respectively, and the copies of HBV DNA decreased by 10(1.77 2.39) (t = 3.13, P < 0.01). Two patients who were treated in combination with lamivudine had complete clinical response. There was no difference in the trial effect between the DC treatment and the other two antiviral methods, and in the efficient rate between the patients whose ALT levels were high before treatment and those whose ALT levels were normal.</p><p><b>CONCLUSION</b>The autologous HBsAg-loaded DCs can effectively suppress HBV replication, reduce virus load in serum, eliminate HBeAg and promote HBeAg/ anti-HBe seroconversion. The patients whose ALT levels are high or normal can response clinically to DCs treatment. DCs in combination with lamivudine can eliminate virus more effectively.</p>

Nanobacteria in serum, bile and gallbladder mucosa of cholecystolithiasis patients / 中华外科杂志

<p><b>OBJECTIVE</b>To find the distribution of nanobacteria in the serum, bile and gallbladder mucosa of cholecystolithiasis patients.</p><p><b>METHODS</b>The infection rate of nanobacteria was identified by ELISA in the serum samples from 338 healthy people and 76 patients with cholecystolithiasis (chi(2) = 0.89, P > 0.05). Nanobacteria were cultured from the bile samples in 57 patients with cholecystolithiasis and 18 non-cholelithiasis patients and identified by immunohistochemical staining and TEM (chi(2) = 29.80, P < 0.05). Forty samples of gallbladder mucosa randomly selected from the 57 cholecystolithiasis patients were identified by immunohistochemical staining and compared with the corresponding bile samples.</p><p><b>RESULTS</b>The infection rate of nanobacteria was 8.0% and 31.6% for the serum samples of the healthy people and cholecystolithiasis patients, respectively. The positive rate of nanobacteria in the bile samples was 61.3% and there was no significant difference in the bile of the cholecystolithiasis patients and the control group (61.4% vs. 61.1%). Fourteen positive patients had infection of nanobacteria in the gallbladder mucosa, submucosa, and calcific field.</p><p><b>CONCLUSIONS</b>The infection rate of nanobacteria was 8% in the serum samples from the healthy people. There are nanobacteria in the serum, bile, and gallbladder mucosa. The infection of the nanobacteria may result in calcification and fibrosis of the gallbladder.</p>

Transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells.</p><p><b>METHODS</b>Enkaryotic expression plasmids inserted HBsAg gene or chimeric HBsAg-HSP70 gene were prepared and transfected into HepG2 cells by means of cationic liposome. mRNA were detected by RT-PCR and proteins expressed in the cells were detected by immunocytochemistry 48 hours later. HBsAg in cultured supernatants and cell lysates were assayed by ELISA.</p><p><b>RESULTS</b>Fusion protein (HBsAg-HSP70) transient expression in HepG2 cells were confirmed by RT-PCR, immunocytochemistry or ELISA, but fusion protein was not assayed in cell cultured supernatants by ELISA.</p><p><b>CONCLUSIONS</b>Transfection of HepG2 cells with a chimeric HBsAg-HSP70 construct leads to express fusion protein, but it does not secrete into cell cultured supernatants.</p>

Study on induction of dendritic cells from myeloid leukemia cell lines and their antitumor immune function / 中国实验血液学杂志

Dendritic cell (DC) plays a key role in antitumor immune response. However, there is a deficiency of DC function in the majority of leukemia patients. It is a novel idea that expanding DC in vitro and enhancing their antitumor immune function and DC-based tumor vaccines may be used as an efficient immune therapy for leukemia. In the project, the condition to induce DC from myeloid leukemia cell lines and its anti-leukemia response were investigated. HL-60, K562 and THP-1 cells were cultured with various combinations of cytokines for inducing DC. The morphologic features were analyzed with optical and electron microscopy. The phenotype of DC was detected by FCM with CD1a, CD40, CD80, CD86, HLA-A, B, C and HLA-DR monoclonal antibodies. The ability of DC stimulating lymphocyte proliferation was observed by allo-mixed lymphocyte reaction using (3)H-TdR incorporation. Cytotoxicity assay was measured by (51)Cr-release method. The level of IL-12 and IFN-gamma in supernatant of DC culture was measured by ELISA. It was proved that the DCs derived from K562, HL-60 and THP-1 cells showed a typical morphology of dendritic cell. The induced cells expressed the surface differentiation antigens of DC. A high expression of phenotypes was found in HL-60-DC and THP-1-DC stimulated by GM-CSF + IL-4 + TNF-gamma and K562-DC with GM-CSF + IL-4 + IL-12. The DCs from the 3 leukemia cell lines stimulated allo-MLR and CTL reaction strongly. Different contents of IL-12 were detected in the supernatants of DC culture and IFN-gamma in the coculture of DC and blood mononuclear cells. It is concluded that the myeloid leukemia cells are able to be induced DCs by cytokines in vitro. The different leukemia cells need different cytokines and cultural conditions. DCs derived from leukemia cells express phenotype of antigen-presenting cells. They have the ability of stimulating T lymphocyte proliferation and inducing CTL reaction to clear leukemia cells, and the DCs secrete IL-12 and increase secretion of IFN-gamma by T cells.