Efficacy and peripheral immunity analysis of allogeneic natural killer cells therapy in patients with hepatocellular carcinoma / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the safety and efficacy of allogeneic natural killer (NK) cells in the treatment of primary hepatocellular carcinoma (HCC), and to elucidate the mechanism of NK cells therapy.@*METHODS@#Twenty-one patients with primary HCC treated with allogeneic NK cells at the Fifth Medical Center of the PLA General Hospital were followed up for 1 year. Peripheral blood mononuclear cells (PBMCs) were isolated from patient-related donors and cultured in vitro for 15 days and infused to the patients in two consecutive days. Clinical data and laboratory data were collected and analyzed, including survival, clinical features, imaging changes, hematology, immunology, and biochemical indicators to evaluate the safety and efficacy of allogeneic NK cell therapy. The changes of peripheral blood lymphocyte subsets after treatment were also analyzed to explore the possible anti-tumor mechanisms.@*RESULTS@#(1) Of the 21 patients with primary HCC, 11 patients were treated once, 5 patients were treated twice, and 5 patients were treated 3 times. After allogeneic NK cells infusion, 10 patients had fever, 1 patient had slight hepatalgia and 1 patient had slight headache, no other adverse events occurred including acute and chronic graft-versus-host disease (GVHD). They resolved spontaneously within 8 hours without other treatment. (2) The total disease control rate was 76.2% during one-year follow-up. Among them, the patients with Barcelona clinic liver cancer (BCLC) stage A had a disease control rate of 100%, stable disease (SD) in 10 cases; BCLC stage B patients had a disease control rate of 60%, partial response (PR) in 1 case, and SD 2 in cases; BCLC stage C patients had a disease control rate of 50%, complete response (CR) in 1 case, and 2 cases of PR. (3) The frequencies of NK cells and CD8+ T cells in peripheral blood were significantly lower than that before at 24 hours after treatment, and the frequencies of CD4+ T cells and CD4/CD8 were significantly higher than the baseline.@*CONCLUSION@#Allogeneic NK cells have good safety and efficacy in the treatment of primary HCC. The anti-tumor effect of the allogeneic NK cells may play an important role in the activation of the patient's natural immune system and delay disease progression, suggesting that allogeneic NK cells combined with sorafenib may be a very effective treatment for advanced HCC, and further large-sample multicenter randomized controlled clinical trials are needed to validate this result.

A Fast Analytical Approach for Determination of 16 Kinds of Mycotoxins in Vegetable Oils Using Stable Isotope Dilution and Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry / 分析化学

A fast, simple and cost-effective UPLC-MS / MS method was established for determination of 16 kinds of mycotoxins in vegetable oils with stable isotope dilution technique. Samples were extracted by acetonitrile-water-acetic acid 84: 15: 1(V/ V) and then diluted using water without any further clean-up steps. The mycotoxins were fully separated on a pentafluorophenyl column. Matrix effects were efficiently compensated by the [ 13 C]-labelled internal standards. The mean recoveries at three different concentration levels ranged from 74. 2％ to 105. 6％ , with RSD varied from 0. 3％ to 13. 9％ . Finally, the method was applied to analyze several kinds of vegetable oil samples. The method was simple, rapid, high sensitive and suitable for the determination of mycotoxins in vegetable oils.

Expressions of CD34 and CD117 in human hepatocellular carcinomas and the clinical significance / 中华肝脏病杂志

To study the expressions of CD34 and CD117 in the tissues of hepatocelluar carcinoma (HCC) and to explore the relationship with clinical pathology and it's evaluation on the prognosis of HCC patients. The expressions of CD34 and CD117 were examined by two-step methods of PV-9000 of immunohistochemistry in 55 HCC cases, 10 liver cirrhotic specimens and 6 normal liver specimens. Clinical-pathological data, tumor recurrent rate and survival rate after hepatectomy were recorded and analyzed with Fisher's Exact Test, Pearson X2 Test, Kaplan-Meier, Log-Rank Test and Cox Regression. The positive expression of CD34 was found in 65.4% of HCC, 20% of cirrhostic liver specimens and 16.7% of normal liver specimens, respectively. Significant differences found among the three groups, and the CD34 expression was significantly associated with vessel embolus (X2 = 4.000, P = 0.046) and the histological grades (X2 = 11.008, P = 0.001). The positive expression of CD117 was 47.3%, 10% and 0% in HCC, cirrhotic liver specimens and normal liver tissues, respectively, and statistical differences esxisted among the three groups. The CD117 expression was dramatically related to the histological grades (X2 = 5.115, P = 0.024) and clinical stages (X2 = 15.459, P = 0.000). Median disease free survival time after hepatectomy was significantly shorter in the group with positive-expression of CD34 (X2 = 4.105, P = 0.043) and CD117 (X2 = 28.023, P = 0.000) than the negative-expressed groups, respectively. Multivariate analysis showed that CD117 expression status, serum AFP levels and the size of tumor were independently prognostic factors for HCC patients. Tthe results demonstrated that CD34 and CD117 might play an important role in liver carcinogenesis and the progression of HCC, and they might potentially serve as markers for HCC prognosis.

Correction of the epicanthal fold and angulus oculi using the Z-epicanthoplasty / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore a new procedure for aesthetic correction of the medial epicanthal fold aim at the etiopathogenesis.</p><p><b>METHODS</b>The new Z-epicanthoplasty devise the upper and inferior margin of angle of eye medial as one angle of the Z.</p><p><b>RESULTS</b>From 2004 to 2006, 129 patients were treated by using the method. Follow-up 6 to 24 months, all patients were satisfied by eliminating the medial epicanthal fold without obvious scar.</p><p><b>CONCLUSIONS</b>The method is more effect than traditionally Z-plasty. Our technique is a simple, advanced procedure that can be performed widely.</p>

Reconstruction of the remnant penis: a 52-case report / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of the combined method of abdominal axial flap transposition and penile elongation for the treatment of the remnant penis.</p><p><b>METHODS</b>Fifty-two cases of the remnant penis treated with the combined method from 1984 April to February 2004 were analyzed retrospectively. Follow-up ranged from 0.5 to 20 years postoperatively.</p><p><b>RESULTS</b>The lengths (both in normal and erectile conditions) and the circumferences of the penis gained after operation were (5.6 +/- 1.4) cm, (6.8 +/- 2.5 cm and (6.9 +/- 2.3) cm respectively. The recovery rates of the sensory function were 94.2% and 100% in the glans (immediately and 3 months after operation) and 32.7%, 51.9% and 75% in the flap area (3, 6 and 12 months postoperatively). The two-point distinguishing sense in the glans and the flap area was (5.1 +/- 0.9) mm and(7.9 +/- 1.3) mm 5 years after operation. Early complications included distant flap necrosis (3 cases), disruption of the wound (2 cases), part necrosis of the skin graft in the abdominal wall (2 cases) and poor contours occurred in 4 cases in the later period because of the thickness of the flaps. All of them were corrected with satisfactory results.</p><p><b>CONCLUSION</b>The combined method of abdominal axial flap transposition and penile elongation was recommendable for the treatment of the remnant penis because of its positive effects and less complications.</p>

Correlation of pathology in chronic hepatitis B to viral markers in serum and hepatic tissue / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the relation of the viral markers in serum and those expressed by hepatocytes to pathological lesions of hepatic tissue in patients with chronic hepatitis B.</p><p><b>METHODS</b>The relation of viral markers including HBsAg, HBsAb, HBeAg, HBeAb, HBcAb and HBV DNA in serum of 647 patients with chronic hepatitis B and HBsAg, HBcAg expressed by hepatocytes in 418 of these patients to pathological lesions of hepatic tissue was determined.</p><p><b>RESULTS</b>Viral markers in serum and those expressed by hepatocytes in patients with chronic hepatitis B were closely correlated with pathological lesions of hepatic tissue.</p><p><b>CONCLUSION</b>The degree of inflammation and fibrosis in hepatic tissue is milder in serum HBsAg, HBeAb, HBcAb positive and HBV DNA negative patients but more serious in those with negative hepatocytic expression of HBsAg and HBcAg. HBV DNA is not significantly associated with pathological lesions of hepatic tissue.</p>

Dynamic expression of matrix metalloproteinase-2, membrane type-matrix metalloproteinase-2 in experimental hepatic fibrosis and its reversal in rat / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the expression dynamics and significance of matrix metalloproteinase-2 (MMP-2) membrane type-matrix metalloproteinase-2 (MT-MMP-2) in hepatic fibrosis and its reversal counterpart.</p><p><b>METHODS</b>An experimental CCl4 induced hepatic fibrosis rat model was established by intraperitoneal administration of carbon tetrachloride for 2, 4, 6, 8, 10 weeks, and normal rats were used as a control group. The immunohistochemical methods and in situ hybridization were used to detect MMP-2,MT-MMP-2 mRNA and related antigens in the liver.</p><p><b>RESULTS</b>MMP-2,MT-MMP-2 mRNA and related antigens were expressed in mesenchymal cells and parts of hepatocytes besides active pathological changes, especially in the fibrous septum and portal area. Expression of MMP-2,MT-MMP-2 mRNA and related antigens were increased in hepatic fibrosis and decreased gradually in its reversal counterpart.</p><p><b>CONCLUSION</b>This study suggested that mesenchymal cells are the main cellular origins of MMPs. The levels of MMP-2 and MT-MMP-2 antigens and gene expression were closely related to hepatic fibrosis. MMP-2 and MT-MMP-2 may play important roles in hepatic fibrosis and its reversal counterpart.</p>

Clinical and pathological analysis on characteristics of primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To explore the clinical and pathological features and the pathogenesis of primary biliary cirrhosis (PBC) in Chinese Mainland.</p><p><b>METHODS</b>30 PBC patients were divided into the early group (Scheuer stage I and II, 19 patients) and the late group (Scheuer stage III and IV, 11 patients). The data of clinics and serology were analyzed, and the pathological features of the liver tissues were characterized. The changes of dendritic cells (DCs) and hepatic stellate cells (HSCs) were studied by immunohistochemistry.</p><p><b>RESULTS</b>In all the PBC patients, the rate of the male to the female was 1 to 5, and the average age was 40.6 years. The mean levels of TBiL, ALP and GGT in the sera were (95.9+-88.5) micromol/L, (537.2+-339.2) U/L, and (582.0+-351.2) U/L, respectively. 73.3% patients showed AMA positive, and the level of GGT was positively correlated with the AMA level according to the result of statistical analysis (r=0.778, P=0.000). The symptoms of jaundice and hepatomegaly were presented more commonly in the late group than those in the early group (chi2=5.182, P<0.05; chi2=13.659, P<0.01, respectively). The main changes of morphology of PBC located in portal tracts. The liver tissues in the early stage of PBC showed the damage of bile ducts and obvious proliferation of small bile ducts. The granulomas, the lymphoid follicles and the foamy cells were found in the liver tissues of PBC (2/19 patients, 12/19 patients, and 10/19 patients in the early stage respectively, while 0/11 patients, 4/11 patients, and 3/11 patients in the late stage respectively). There was significant difference between the early stage and the late stage in presence of the lymphoid follicles and the foamy cells (t=4.489, P<0.05; t=4.019, P<0.05, respectively). The biliary pigmentary particles were mainly accumulated in the liver cells around the portal tracts in 90.0% PBC patients, and the accumulation of copper and iron increased, compared with that in normal specimens. The DCs and HSCs located mainly in the portal tracts, especially around the damaged bile ducts.</p><p><b>CONCLUSIONS</b>There are some clinical and pathological characteristics in the patients with PBC. The level of AMA has no direct relationship with the level of transaminase or bilirubin. The proliferated bile ductules may express the antigens which maybe the target of immune attack. As an antigen-presenting cell, DCs may play an important role in the pathogenesis of PBC.</p>

Cut-off period of subclassification and pathological features of severe hepatitis based on clinical and pathological analyses / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To explore the cut-off period of subclassification and pathological features of severe hepatitis (SH).</p><p><b>METHODS</b>Based on combined clinical and pathological analyses, the complete clinical and biopsy or autopsy liver tissues data from 196 cases of patients with severe hepatitis were investigated. Meanwhile, proliferative hepatocytes, cholangioepithelia and collagens were identified by a panel of monoclonal antibodies such as those against albumin, cytokeratin 18,19 and collagen I, III with immunohistochemical method.</p><p><b>RESULTS</b>The clinical and pathological analyses indicated the cut-off periods of acute, subacute and chronic SH (ASH,SSH and CSH) were (13.4+/-7.2) d, (77.4+/-69.3) d and (80.5+/-63.2) d, respectively. Among all SH cases, one case of ASH patient presented clinical manifestation and pathological changes of ASH for 21 days, however, one patient with SSH was demonstrated 12 day course by histological examination. The time of cut-off period between ASH and SSH in child cases was shorter than that in adult cases. Histologically, ASH liver tissues showed massive and/or submassive necrosis caused by one attack, with congestive sinusoid frameworks and proliferative cholangioepithelium-like hepatocytes, while SSH liver tissues presented combined fresh and old submassive or massive necrosis caused by multiple attacks, accompanied by obviously proliferative bile ducts and sinusoid framework collapse.However, the pathological changes of CSH showed ASH- or SSH-like lesions on the background of chronic liver injury.</p><p><b>CONCLUSION</b>Our data indicated that the cut-off period between ASH and SSH is in accordance with the Scheme of Viral Hepatitis Prevention and Therapy, China, published in 2000, but excluded a part of child SH cases. In our study, the authors found a few pathological features in ASH and SSH.</p>

Clinical pathology and pathogenesis of severe acute respiratory syndrome / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS.</p><p><b>METHODS</b>Pathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry.</p><p><b>RESULTS</b>In total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens.</p><p><b>CONCLUSIONS</b>Histologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury.</p>