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To investigate the effect of baicalein on polymicrobial sepsis-induced immune dysfunction and organ injury.A sepsis model was induced in Sprague-Dawley rats via caecal ligation and puncture (CLP). Specific pathogen free rats were randomly divided into a sham group, CLP group and CLP + baicalein (Bai) group (n=16 each). Rats in the CLP + Bai group were intravenously injected with baicalein (20 mg/kg) at 1 and 10 h after CLP. Survival rate, bacterial load, and organ damage were assessed. Then each group was evaluated at 6, 12, and 24 h to investigate the effect of baicalein on immune cells and inflammatory cytokines in septic rats.Baicalein treatment significantly improved the survival of septic rats, decreased the bacterial burden, and moderated tissue damage (spleen, liver, and lung), as observed by haematoxylin and eosin staining. Septic rats treated with baicalein had strikingly increased proportions of CD3Baicalein may be an effective immunomodulator that attenuates overwhelming inflammatory responses in severe abdominal sepsis.

Background The aim of the study was to evaluate the current opinion and clinical decision-making process of international pancreatologists, and to systematically identify key study questions regarding the diagnosis and treatment of chronic pancreatitis (CP) for future research. Methods An online survey, including questions regarding the diagnosis and treatment of CP and several controversial clinical case vignettes, was send by e-mail to members of various international pancreatic associations: IHPBA, APA, EPC, ESGE and DPSG. Results A total of 288 pancreatologists, 56% surgeons and 44% gastroenterologists, from at least 47 countries, participated in the survey. About half (48%) of the specialists used a classification tool for the diagnosis of CP, including the Mayo Clinic (28%), Mannheim (25%), or Buchler (25%) tools. Overall, CT was the preferred imaging modality for evaluation of an enlarged pancreatic head (59%), pseudocyst (55%), calcifications (75%), and peripancreatic fat infiltration (68%). MRI was preferred for assessment of main pancreatic duct (MPD) abnormalities (60%). Total pancreatectomy with auto-islet transplantation was the preferred treatment in patients with parenchymal calcifications without MPD abnormalities and in patients with refractory pain despite maximal medical, endoscopic, and surgical treatment. In patients with an enlarged pancreatic head, 58% preferred initial surgery (PPPD) versus 42% initial endoscopy. In patients with a dilated MPD and intraductal stones 56% preferred initial endoscopic ± ESWL treatment and 29% preferred initial surgical treatment. Conclusion Worldwide, clinical decision-making in CP is largely based on local expertise, beliefs and disbeliefs. Further development of evidence-based guidelines based on well designed (randomized) studies is strongly encouraged.

BackgroundHypersplenism is a common consequence of liver cirrhosis and a risk factor of hepatocellular carcinoma (HCC) development. The objective of the current study was to identify whether splenectomy for treatment of hypersplenism has any impact on risk of HCC. ­­MethodsPatients who underwent splenectomy for hypersplenism secondary to liver cirrhosis and portal hypertension between 2008 and 2012 were included from seven University Hospitals in China, whereas patients receiving medication treatments for liver cirrhosis and portal hypertension (non-splenectomy) at this time period were also included as control. ResultsA total of 871 patients with liver cirrhosis and hypertension were included synchronously from 7 tertiary hospitals. Among them, 407 patients had a history of splenectomy for hypersplenism, whereas 464 patients who received medical treatment but not splenectomy (non-splenectomy group). The cumulative incidence of HCC development at 1, 3, 5 and 7 years were 1%, 6%, 11% and 16% in splenectomy group, and 2%, 10%, 17% and 24% in non-splenectomy group in the unadjusted cohort (Breslow test = 7.7, p=0.005). Consistently, in the matched cohort, the cumulative rates of HCC diagnosis at 1, 3, 5 and 7 years were 1%, 6%, 7% and 15% in splenectomy group, and 1%, 6%, 15% and 23% in non-splenectomy group (Breslow test = 4.9, p=0.028). On multivariable analysis, splenectomy was independently associated with decreased risk of HCC development (HR 0.55, 95% CI, 0.32-0.95, p=0.031). ConclusionSplenectomy for treatment of hypersplenism may decrease the risk of HCC development.

Previous studies using contrast-enhanced imaging for glioma isocitrate dehydrogenase (IDH) mutation assessment showed promising yet inconsistent results, and this study attempts to explore this problem by using an advanced tracer kinetic model, the distributed parameter model (DP). Fifty-five patients with glioma examined using dynamic contrast-enhanced imaging sequence at a 3.0 T scanner were retrospectively reviewed. The imaging data were processed using DP, yielding the following parameters: blood flow F, permeability-surface area product PS, fractional volume of interstitial space Ve, fractional volume of intravascular space Vp, and extraction ratio E. The results were compared with the Tofts model. The Wilcoxon test and boxplot were utilized for assessment of differences of model parameters between IDH-mutant and IDH-wildtype gliomas. Spearman correlation r was employed to investigate the relationship between DP and Tofts parameters. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis and quantified using the area under the ROC curve (AUC). Results showed that IDH-mutant gliomas were significantly lower in F ( P = 0.018), PS ( P < 0.001 ), Vp ( P < 0.001 ), E ( P < 0.001 ), and Ve ( P = 0.002) than IDH-wildtype gliomas. In differentiating IDH-mutant and IDH-wildtype gliomas, Vp had the best performance (AUC = 0.92), and the AUCs of PS and E were 0.82 and 0.80, respectively. In comparison, Tofts parameters were lower in Ktrans ( P = 0.013) and Ve ( P < 0.001 ) for IDH-mutant gliomas. No significant difference was observed in Kep ( P = 0.525). The AUCs of Ktrans, Ve, and Kep were 0.69, 0.79, and 0.55, respectively. Tofts-derived Ve showed a strong correlation with DP-derived Ve (r > 0.9, P < 0.001 ). Ktrans showed a weak correlation with F (r P > 0.16) and a very weak correlation with PS (r P > 0.8), both of which were not statistically significant. The findings by DP revealed a tissue environment with lower vascularity, lower vessel permeability, and lower blood flow in IDH-mutant than in IDH-wildtype gliomas, being hostile to cellular differentiation of oncogenic effects in IDH-mutated gliomas, which might help to explain the better outcomes in IDH-mutated glioma patients than in glioma patients of IDH-wildtype. The advantage of DP over Tofts in glioma DCE data analysis was demonstrated in terms of clearer elucidation of tissue microenvironment and better performance in IDH mutation assessment.

Additional file 1.

Myeloid-derived suppressor cells (MDSCs) expand in tumor-bearing host. They suppress anti-tumor immune response and promote tumor growth. Chemokines play a vital role in recruiting MDSCs into tumor tissue. They can also induce the generation of MDSCs in the bone marrow, maintain their suppressive activity, and promote their proliferation and differentiation. Here, we review CCL2/CCL12-CCR2, CCL3/4/5-CCR5, CCL15-CCR1, CX3CL1/CCL26-CX3CR1, CXCL5/2/1-CXCR2, CXCL8-CXCR1/2, CCL21-CCR7, CXCL13-CXCR5 signaling pathways, their role in MDSCs recruitment to tumor tissue, and their correlation with tumor development, metastasis and prognosis. Targeting chemokines and their receptors may serve as a promising strategy in immunotherapy, especially combined with other strategies such as chemotherapy, cyclin-dependent kinase or immune checkpoints inhibitors.

ObjectiveTo explore the protection and molecular mechanism of histone deacetylase inhibitors (HDACIs) on the spleen of rats with hemorrhagic shock.MethodsA total of 60 SPF male SD rats were selected for the modeling of severe hemorrhagic shock using the method of arterial and venous cannulation with the time-divided bleeding. The measurement of mean arterial blood pressure and blood lactic acid was used to verify the modeling. The modeled rats were randomly divided into shock group, shock + suberoylanilide hydroxamic acid (SAHA) group, shock + autogenous transfusion group and shock + SAHA + autogenous transfusion group. Three hours after the treatment, the spleen of rats was collected and TUNEL method was employed to detect the apoptosis of spleen cells in each group. The statistical analysis was performed. Afterwards, real-time PCR and western blot were employed to detect the expression of BCL-2, BAX and caspass3 in the spleen of rats in each group.ResultsA total of 53 rats had successful modeling of severe hemorrhagic shock, with success rate of 88%. Cell apoptosis in the severe hemorrhagic model group was the most serious. After the intervention of HDACIs and the autogenous transfusion, the tissue injury was a bit recovered. Cell apoptosis was least in the shock + SAHA + autogenous transfusion group (P

Relationship between dose and response. Relationship between CAR+ cell dose and complete response. (DOCX 80Â kb)

Genomic copies of LCAR-B38M. Copies of LCAR-B38M per microgram of genomic DNA. (DOCX 90Â kb)

Relationship between BCMA expression and clinical response, progression-free survival, and overall survival. Overall response rate, progression-free survival, and overall survival by BCMA expression. (DOCX 15Â kb)

LCAR-B38M lentiviral construct. Figure depicting the LCAR-B38M lentiviral construct. (DOCX 17Â kb)

Interleukin-6 levels by cytokine release syndrome grade. Peak IL-6 levels by CRS grade. (DOCX 47Â kb)

Immunoglobulin levels following LCAR-B38M infusion. Levels of serum IgA, IgG, lambda chain, and kappa chain following CAR T cell infusion. (DOCX 281Â kb)

Pituicytoma is a distinct low-grade glioma arising from pituicytes of the neurohypophysis and infundibulum. Due to the rarity of this tumor, most of the reported cases in the literature were originally misdiagnosed as pituitary adenoma, meningiomas, and craniopharyngioma. It is of vital importance to accurately identify this tumor because this tumor, unlike pituitary adenomas, is prone to heavy bleeding during the surgical resection.[1]

Choroid plexus epithelial cells secrete numerous biologically active neurotrophic factors, which may be beneficial to the transplantation site. Encapsulated cells are often used in tissue transplantation. The present study was conducted to investigate the effect of encapsulation on the secretory function of choroid plexus epithelial cells. Neonatal rat choroid plexus epithelial cells were primarily cultured. After 9 days of culture, the cells were distributed into two groups, and one group of cells was encapsulated in vitro. The initial culture conditions such as cell numbers and medium volumes were the same. Supernatants in the free and encapsulated choroid plexus epithelial cells were collected at the time points of day 1 through day 7. Quantitative determination of the BDNF and GDNF levels was performed by enzyme-linked immunosorbent assay to assess the secretory function of the cells in the two forms. Statistical analyses were performed using a Student t test. P

BACKGROUND For patients with stage IV gastric cancer, it is unclear whether splenectomy combined with palliative surgery is needed to reduce tumor load and relieve symptoms. The objective of the present study was to investigate the effect of splenectomy combined with palliative resection for stage IV gastric carcinoma on immunological dysfunction and patient prognosis. MATERIAL AND METHODS We retrospectively analyzed medical records of 106 stage IV gastric cancer patients who underwent palliative surgery; of these, 49 patients were treated with palliative resection for gastric carcinoma combined with splenectomy, while the other 57 patients retained their spleens. The immunologic function and prognosis in these 2 groups were examined and compared. RESULTS The immune function of patients in the group that retained their spleens was better later in the postoperative course than in the resection group. The groups did not show statistically significant differences in postoperative infectious complications, median survival time, and survival rate; however, the average postoperative hospitalization time of patients in the retained group was significantly shorter. CONCLUSIONS Splenectomy combined with gastric cancer resection did not improve the prognosis of the patients; patients who retained their spleens had faster recovery and improved immune function. However, whether retaining the spleen is an independent factor improving the prognosis needs further investigation.

To investigate the expression of p16INK4a protein in breast cancer and analyze its clinical significance.A total of 132 surgical specimens of primary breast cancer obtained between 2014 and 2015 were examined for expressions of ER, PR, CK5/6, Her-2 and p16INK4a proteins using immunohistochemistry.The breast cancer samples were classified into 5 molecular subtypes, namely Luminal A (58 cases), Luminal B (32 cases), Her-2-positive (21 cases), basal-like (12 cases) and normal-like (9 cases) types. p16INK4a expression was negative in 7/132 (5.30%) cases, weakly positive in 15/132 (11.36%) cases, positive in 40/132 (30.30%) cases, and strongly positive in 70/132 (53.03%) cases. When categorizing negative and weakly positive cases into negative group and the positive and strongly positive cases into positive group, the total negative and positive expression rates of p16INK4a were 16.67% (22/132) and 83.33% (110/132) in the carcinoma tissues. Statistical analysis showed the expression intensity of p16INK4a differed significantly between the age groups (P0.05) but was not significantly correlated with ER, PR, Her-2, molecular subtypes or metastasis of the tumors.The compensatory high expression of p16INK4a is the main mechanism of cell cycle deregulation in invasive breast cancer and can be an important specific molecular marker for invasive breast cancer.

Early-onset familial Alzheimer's disease (EOFAD) is characterized by the onset of dementia symptoms before 65 years, positive family history, high genetic predisposition, and an autosomal dominant inheritance. We aimed to investigate mutations and to characterize phenotypes in Chinese EOFAD families. Detailed clinical assessments and genetic screening for mutations in the presenilin 1 (PSEN1), presenilin 2, amyloid precursor protein, and APOE genes were carried out in 4 EOFAD families. Two PSEN1 mutations (p.R352C and p.M233L) were identified in 2 EOFAD families, respectively. Mutation p.M233L was associated with prominent very early onset, rapidly progressive dementia, and neurologic symptoms, whereas p.R352C was associated with a progressive dementia, psychiatric syndrome, and chronic disease course. Both mutations are predicted to be pathogenic. Our results showed that mutations in PSEN1 gene might be common in Chinese EOFAD families.

The present study was aimed to investigate the protective effects of ginsenoside Rg1 (GRg1), an important component of ginseng, in oxygen-glucose deprivation/reperfusion (OGDR) and to elucidate the related mechanisms. PC12 cells were used as the model of OGDR. GRg1 administration was started 12 h before OGD and lasted for 12 h. After OGD, the cells were incubated in drug-free and full culture medium under normoxic condition for 24 h. Cell viability was then measured using MTT assay. Cell morphology was studied under a microscope. The expressions of survivin, caspase-3 and Terminal dUTP nick-end labeling (TUNEL) were measured by immunocytology. Results showed that pretreatment with GRg1 significantly increased the viability and survivin expression, and decreased the expressions of caspase-3 and TUNEL in a dose-dependent manner. In addition, it dramatically increased the number of cells and improved the cellular morphology. These results demonstrate the effect of GRg1 in preventing OGDR-induced PC12 cell apoptosis and partly reveal the mechanisms of the protective effect. It is suggested that GRg1 has potential beneficial effects in ischemic diseases.

Oxymatrine is an alkaloid obtained primarily from Sophora roots and has been shown to show anticancer effects in various cancers. However, the cellular and molecular effects of this agent on cervical cancer have been poorly characterized. Here, we investigated the antitumor effect of oxymatrine on a human cervical cancer cell line (HeLa). Our results showed that application of oxymatrine significantly inhibited the cell growth and tumorigenesis in a dose-dependent manner and induced apoptosis through caspase-dependent pathways as determined using flow cytometry and TUNEL staining analysis. To define the proteins potentially related to the mechanisms of action, proteomic analysis was utilized to detect proteins altered by oxymatrine. As the downregulated gene, inosine monophosphate dehydrogenase type II (IMPDH2) was responsible for oxymatrine-induced mitochondrial-related apoptosis. Moreover, oxymatrine depleted intracellular guanosine 5'-triphosphate (GTP) levels by effective IMPDH inhibition. Functional analyses further showed that oxymatrine and tiazofurin, an inhibitor of IMPDH2, sensitized resistant HeLa/DDP cells to cisplatin. In addition, the expression of IMPDH2 in cervical cancer was significantly higher than that in the normal cervical epithelium. Taken together, these findings suggest that targeting of IMPDH2 by potential pharmacological inhibitors, oxymatrine in combination with chemotherapy, might be a promising means of overcoming chemoresistance in cervical cancer with high IMPDH2 expression, and may thus provide new insights into the mechanism of oxyamtrine-induced anticancer effects.

To establish a method for the culture of primary choroidal epithelial cells.This descriptive experimental study was carried out in Xi`an Jiaotong University, Xi`an, China from September 2009 to August 2012. Choroidal epithelial cells were isolated from the choroid plexus tissues of the lateral ventricles from neonatal rats (n=36). The tissues were dissociated into small cell aggregates by a mechanical method, and cultured on plastic culture dishes containing Dulbecco`s modified Eagle`s medium with 10% fetal bovine serum and 10 ng/ml epidermal growth factor at 37 degrees C in an incubator with 5% humidified carbon dioxide. The cultured cells were examined by phase contrast microscope, electron microscopy, and immunocytochemistry.The cells showed typical morphologic characteristics of epithelial phenotypes with a cobblestone appearance in monolayer 7-9 days post-seeding. The electron microscopy spotted typical choroidal epithelial cells with microvilli on the cytomembrane, organelles in the cytoplasm, and tight junctions welding 2 adjacent cells. They were positive against anti-transthyretin immunostaining.This culture technique, which does not require complex equipment and operation skills, might be a simple and efficient method for obtaining choroidal epithelial cells in sufficient number and purity from mixed primary cultures of rat tissue.

Background: The choroid plexuses, which could secrete a number of neurotrophins, have recently been used in transplantation in central nervous system diseases. Aims: To study the mechanism of nerve regeneration in the central nervous system by grafting choroid plexus tissues. Study Design: Animal experimentation. Methods: The choroid plexuses from the lateral ventricles of neonatal rats were cultured in adherent culture, and immunocytochemical methods were used to analyse the progenitor cells on days 2, 6, and 10 after seeding. Results: Expression of both nestin and glial fibrillary acidic protein was observed in small cell aggregates on day 2 in primary culture. Most of the nestin-positive cells on day 6 were immunoreactive to glial fibrillary acidic protein antibody. No cells expressing nestin or glial fibrillary acidic protein were seen on day 10. Conclusion: These experimental results indicate that the choroid plexus contains a specific cell population – progenitor cells. Under in vitro experimental conditions, the progenitor cells differentiated into choroid plexus epithelial cells but did not form neurons or astrocytes. Turkish Başlık:Farklılaşma Kabiliyeti Olan Progenitör Hücreler İçeren Yenidoğan Ratların Koroid Pleksusunun Primer Kültürü Anahtar Kelimeler:Koroid pleksus, primer kültür, progenitör hücreler, farklılaşma, rat Arkaplan: Bir dizi nörotrofin salgılayabilen koroid pleksus, son zamanlarda merkezi sinir sistemi hastalıklarında transplantasyonda kullanılmaktadır. Amaç: Koroid pleksus dokularının greftlenmesi ile merkezi sinir sisteminde sinir rejenerasyonunun mekanizmasını incelemek. Çalışma Tasarımı: Hayvan deneyi Yöntemler: Yenidoğan ratların lateral ventrikülünden alınan koroid pleksuslar yapışık (adherent) kültür ile çoğaltıldı ve immünositokimyasal metotlar ekim sonrası 2., 6. ve 10. günlerde progenitör hücrelerin analizi için kullanıldı. Bulgular: Primer kültürde 2. günde küçük hücre kümelerinde hem nestin hem de glial fibriler asidik protein ekspresyonu gözlendi. 6. Günde nestin-pozitif hücrelerin çoğu, glial fibriler asidik protein antikoruna karşı immunoreaktif idi. 10. Günde nestin veya glial fibriler asidik protein eksprese eden hücre görülmedi. Sonuç: Bu deneysel sonuçlar, koroid pleksusun spesifik bir hücre popülasyonu-progenitör hücreleri içerdiğini göstermektedir. In vitro deneysel koşullar altında, progenitör hücreler koroid pleksus epitel hücrelerine farklılaşmış ancak nöronları veya astrositleri oluşturmamıştır.

It is widely accepted that intersexual differences occur in cognitive domains, e.g., in spatial learning and memory. The hippocampus plays important roles in the consolidation of information from short-term memory to long-term memory and spatial navigation. However, it still remains unknown whether the hippocampal proteomic profiling differs between males and females. In this study, we investigated the intersexual differences in protein expression of hippocampi using the two-dimensional electrophoresis analysis. In all, 33 differentially expressed proteins were characterized by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry and validated by Western-blotting analysis. In line with Western-blotting validation, the proteomic identification revealed the overexpression of glial fibrillary acidic protein in female rats' hippocampi, and the overexpression of both creatine kinase B-type and DRP-2 in male rats' hippocampi. The intersexual differences in hippocampal proteomic profiling are probably closely related to those in spatial learning and memory abilities.

AIM: To identify discriminating protein patterns in serum samples among non‐small cell lung cancer (NSCLC), chronic obstructive pulmonary disease (COPD), pneumonia, and healthy controls. To discover specific low molecular weight (LMW) serum peptidome biomarkers and establish a diagnostic pattern for NSCLCby using proteomic technology. METHODS: We used magnetic bead‐based separation followed by matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) to identify patients with NSCLC, COPD, and pneumonia. A total of 154 serum samples were analyzed in this study, among which there were 60 serum samples from NSCLC patients, 30 from patients with other lung‐related diseases (16 pneumonia patients and 14 patients with COPD) as disease controls, and 64 from healthy volunteers as healthy control. The mass spectra, analyzed using ClinProTools software, distinguished between cancer patients and healthy individuals based on GA algorithm model. RESULTS: In this study, we generated numerous discriminating m/z peaks as well as disease‐specific discrimination peaks. A set of five potential biomarkers (m/z: 7,763.24, 1,012.61, 4,153.16, 1,450.55, and 2,878.89) could be used as the diagnostic biomarkers to distinguish NSCLCpatients from healthy controls. In the training set, patients with NSCLC could be identified with sensitivity of 97.5% and specificity of 98.8%. Similar results were obtained in the testing set, showing 80.7% sensitivity and 91.2% specificity. CONCLUSION: Our study demonstrated that a combined application of magnetic beads with MALDI‐TOF MS technique was suitable for identification of serum biomarkers for NSCLC. J. Clin. Lab. Anal. 26:148‐154, 2012. © 2012 Wiley Periodicals, Inc.

To investigate the roles of STAT1 and STAT2 in growth inhibition induced by phosphatidylethanolamine (PE) in human hepatoma HepG2 cells.The growth of HepG2 cells exposed to 0.125, 0.25, 0.5 and 1.0 mmol/L PE was assessed by MTT assay, and the expressions of STAT1 and STAT2 were analyzed using immunocytochemical assay.PE inhibited the growth of HepG2 cells in a dose-dependent manner and increased the expression of STAT1 and STAT2 in comparison with those in the control group. AG490, an inhibitor of JAKs, partially reversed PE-induced growth inhibition of HepG2 cells.STAT1 and STAT2 are involved in the growth inhibition of human hepatoma HepG2 cells induced by PE.

It is generally accepted that spleen plays a complex role in the tumor immunity, which would change in the different periods of cancer. In this study, we investigated the changes in the function of splenic macrophage (Mphi) in different stages of liver cancer induced by diethylnitrosamine (DEN) in rats. The aim was to support the characteristics of "two-way" and "phase" of spleen in tumor immunity.The model of pulmonary metastasis of liver cancer was established in forty male SD rats by DEN. In the 8th, 13th and 16th week, 10 rats were randomly chosen and sacrificed, and divided into cirrhosis, liver cancer and pulmonary metastasis groups depending on the pathological result, respectively. The other 10 rats were taken as control group. The Mphi was isolated by anchoring cultivation. The changes in ultrastructure, phagocytosis, cytokine secretion, antigen processing and presenting, and viability of splenic Mphi were detected by transmission electron microscopy, Vybrant(TM) Phagocytosis Assay, DQ(TM) Ovalbumin, and rat TNF-alpha ELISpot kits.Under the electron microscope, the Mphi in the control group had some pseudopodium-like prominences, and mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, and phagocytized RBC. In the liver cirrhosis and liver cancer group, Mphi had more prominences, meanwhile much more mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, especially in the liver cancer group. In the pulmonary metastasis group, the Mphi was swelling, with few organelle. As compared to the control group, the function of splenic Mphi increased in cirrhosis and cancer groups, but decreased in metastasis group (phagocytosis rate: (84.7 +/- 1.9)%, (89.5 +/- 3.1)%, and (36.0 +/- 2.6)% vs (75.6 +/- 1.7)%, P0.05, P0.01; viability: (1.53 +/- 0.15)%, (1. +/- 0.14)%, and (1.12 +/- 0.29)% vs (1.48 +/- 0.17)%, P0.05, P0.01; TNF-alpha secretion: (741.0 +/- 52.9)%, (1126.2 +/- 174.5)%, and (313.8 +/- 50.8)% vs (626.6 +/- 24.6)%, P0.05, P0.01; positive cell rate of antigen processing and presenting: (24.03 +/- 1.87)%, (27.95 +/- 2.63)%, and (10.46 +/- 2.16)% vs (16.45 +/- 1.86)%, P0.01).In the stage of cirrhosis and early cancer, the immune functions of splenic Mphi were reinforced. It may promote the non-specificity tumor immunity. On opposite, in the stage of pulmonary metastasis, the immune functions of splenic Mphi were impaired. It may lead to the decrease of tumor immunity.

Hepatocellular carcinoma (HCC) is an aggressive cancer with a dismal outcome largely due to metastasis and postsurgical recurrence. Thus, the inhibition of invasion and metastasis is of great importance in its therapies. Medicinal plants or ethnopharmacology used in folklore medicine continue to be an important source of discovery and development of novel or potential therapeutic agents for treatment of cancer. Chrysanthemum indicum, one of the medicinal plants or ethnopharmacology, is being used for treatment of many diseases including cancer. However, this plant molecular mechanisms underlining the anti-metastatic effects have not been well documented. In this study, Chrysanthemum indicum ethanolic extract (CIE) significantly suppressed proliferation and invasion of MHCC97H cells, one of the HCC cell lines with high metastatic potential, in a dose-dependent manner. CIE markedly decreased MMP-2 and MMP-9 expression, increased simultaneously TIMP-1, and TIMP-2 expression further restoring their balance in the cancer cells. The present study indicates that CIE reduced MHCC97H cell metastatic capability, in part at least, through decrease of the MMP expression, simultaneous increase of the TIMP expression, further restoring their balance as therapeutic target in HCC. It is suggested that Chrysanthemum indicum is a potential novel therapeutic medicinal plant for treatment of HCC or cancer invasion and metastasis.

To investigate the changes in function of splenic macrophages of hypersplenism due to portal hypertension (PH), and to provide experimental evidence for exploring the immune function of spleen in PH.Twelve patients with hypersplenism due to PH and four patients with traumatic rupture of spleen, from September 2005 to March 2006, were enrolled into PH group and control group, respectively. Splenic M phi were isolated and purified by anchoring cultivation from all the patients, and were resuspended by RPMI-1640. Phagocytosis, cytokine secretion and antigen processing and presenting of splenic M phi were detected by Vybrant Phagocytosis Assay, the human TNF-alpha Elispot kits and DQ ovalbumin.Compare to the normal splenic M phi, the phagocytosis rate, antigen presentation positive cells and secretion positive cells, were all significantly increased in PH splenic M phi (86.4 +/- 7.1 vs. 61.8 +/- 4.1, 26.3 +/- 1.6 vs. 15.6 +/- 1.8, 387.0 +/- 24.3 vs. 240.3 +/- 13.0, P0.01).The phagocytosis, cytokine secretion and antigen processing and presenting of splenic M phi in PH spleen were all significantly increased, and the M phi retained at activated state. It means that the PH spleen still possessed the immune function, but these functions might be in disorder. It still needs more research to get the precious evaluation for immune function in the PH spleen.

Stromal derived factor-1 (SDF-1) is an efficacious leukocyte chemoattractant, which can attract lymphocytes and mononuclear cells from bloodstream into the site of inflammation. Emodin, an anthraquinone derivative from Radix et Rhizoma Rhei, and baicalein, a flavone from Scutellaria baicalensis Georgi, both have been reported to possess anti-inflammatory activities. The expression pattern of SDF-1 in experimental acute pancreatitis (AP) and the effect of emodin or baicalein on that are not well defined. The present study aimed to investigate the effects of emodin and baicalein on pancreatic myeloperoxidase (MPO) activity (reflecting leukocyte sequestration) and cytokine production, as well as tissue SDF-1 expression in the setting of AP.A rat model of AP was induced by administration of 5% sodium taurocholate through the biliopancreatic duct. The level of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and MPO in the pancreas, and serum amylase were tested by immunohistochemistry, ELISA and chromatometry. The expressions of SDF-1 alpha and SDF-1 beta were detected by real-time PCR, Western blotting, and immunohistochemistry.Combination of emodin and baicalein significantly reduced pancreatic TNF-alpha, IL-6 and MPO, and also inhibited pancreatic SDF-1 expression.The inhibition of SDF-1 expression by emodin and baicalein might contribute, in part at least, to the amelioration of pancreatic inflammation. The present study also shows benefits of simultaneous treatment of AP.

Chinese medicine plays an important role in hepatoprotective treatment. This study was conducted to investigate the protective effects of emodin and astragalus polysaccharides (APS) in a rat model of chronic hepatic injury.Chronic hepatic injury was induced by hypodermic injection of an olive oil solution containing 40% carbon tetrachloride (CCl(4)) twice a week, in addition to a diet of 79.5% maizena, 20% fat, 0.5% cholesterol, and 10% alcohol in the drinking water ad libitum for 12 weeks. Meanwhile, the rats were exposed to different concentrations of emodin (40 mg x kg(-1) x d(-1)), APS (200 mg x kg(-1) x d(-1)), combination drug (emodin 40 mg x kg(-1) x d(-1) combined with APS 200 mg x kg(-1) x d(-1)) and colchicine (0.1 mg x kg(-1) x d(-1)) in parallel by oral gavage (once a day for 12 weeks). At the end of 12 weeks, blood serum and liver tissue were taken. Serum was collected to determine the levels of total bilirubin (TBIL), alanine transaminase (ALT), aspartate transaminose (AST), and albumin (ALB). Liver and spleen indexes were assayed, followed by the measurements of the liver associated enzyme superoxide dismutase (SOD) and malondialdehyde (MDA). Histopathological changes were studied using optical microscopy.Splenohepatomegalia was alleviated and serum levels of TBIL and ALT were reduced in the groups treated with emodin and APS when compared to the control group. In addition, the ALB level in the APS and combination groups was higher. Similarly, the SOD activity of liver homogenates was significantly higher in the groups treated with emodin and APS, while administration of the herbal derivatives prevented the elevation in MDA levels. Histological analysis showed that the APS and combination groups significantly ameliorated the hepatic injury.Co-administration of emodin and APS demonstrated a synergistic action in reducing ALT and restoring ALB in the serum from a rat model of chronic hepatic injury. Emodin and APS may ameliorate the CCl(4)-induced hepatic injury in rats by elevating antioxidant-enzyme activities and reducing lipid peroxidation.

To study the mechanism of hepatic carcinoma cell apoptosis induced by small interfering RNA (siRNA)-mediated nuclear factor-kappaB (NF-kappaB) P65 silencing.Hepatic carcinoma SMMC-7721 cells were exposed to liposome-mediated transfection with NF-kappaB P65 siRNA synthesized by in vitro transcription, and the cells with empty liposome transfection and those without particular treatment served as the control groups. The expression of NF-kappaB P65 in the cells was detected by Western blotting, the cell viability examined by MTT assay, and the cell apoptosis assessed by flow cytometry. Immunohistochemistry was used to examine the expressions of Bcl-2 and Bax.siRNA transfection significantly inhibited the expression of NF-kappaB P65 in SMMC-7721cells, with inhibition rates of 64.74% compared with the untreated cells and of 34.52% compared with the liposome-treated cells. The siRNA-treated SMMC-7721 cells also exhibited significant decrease in cell proliferation by 33.39% and 27.23% in comparison with the untreated and liposome-treated cells, respectively. NF-kappaB P65 siRNA induced obvious cell apoptosis with down-regulated Bcl-2 and up-regulated Bax expressions.NF-kappaB p65 siRNA can induce SMMC-7721 cell apoptosis via the Bcl-2/Bax pathway.

To investigate the effect of matrine injections on invasion and metastasis of gastric carcinoma SGC-7901 cells in vitro.MTT assay was used to examine the effect of matrine injections on proliferation of SGC-7901 cells after 24, 48, 72, 96 hours treatment; Transwell chamber assay was performed to determine the effect of matrine injection on invasion and migratory capacity of the cells; Effect on adhesion potential of SGC-7901 cells was tested by cell matrigel adhesion assay.Matrine injec-tions could inhibit the proliferation of SGC-7901 cells, with obvious dose-dependent and time-dependent effects. Matrine injections sig-nificantly inhibited adhesion, invasion and migration capacity of SGC-7901 cells in vitro. The inhibitory rate of them after treatment with 1.5 mg/ml matrine injections for 24 h were (45.32 +/- 3.10)%, (32.66 +/- 2.82)%, (38.35 +/- 3.41)% respectively. After treat-ment of matrine injections (1.5 mg/ml)for 24 h, the expression level of CD44(V6) in SGC-7901 cells was decreased compared with the untreated group.Matrine injections can inhibit the migration, invasion and adhesion capacity of SGC-7901 cells in vitro. The inhibition effect may be related to down-regulating the expression of CD44(V6) protein.

To investigate the relationship between HER-2 expression and the efficacy of neoadjuvant chemotherapy in local advanced breast cancer.Different neoadjuvant chemotherapy regimens, namely CMF, CEF, and NEF, were administered in 132 patients with local advanced breast cancer for 2 cycles, each lasting for 28 days. According to the criteria recommended by WHO, the efficacy and safety of the regimens were evaluated after two cycles of neoadjuvant chemotherapy. HER-2 expression was examined by immunohistochemistry using specific monoclonal antibodies before chemotherapy and after surgery.The overall response rate (RR) of CMF, CEF, and NEF regimens were 39.5% (17/43), 54.3% (25/46) and 72.1% (31/43), with incidence of leukopenia of 34.9% (15/43), 58.7% (27/46) and 60.5% (26/43), respectively. Other adverse effects including decreased hemoglobin (Hb) level, thrombocytopenia, gastrointestinal irritation and alopecia were similar between the 3 groups (P0.05). No significant variation in HER-2 expression occurred after administration of the 3 regimens. The overall RR to CMF regimen in HER-2-negative breast cancer patients was significantly higher than that in HER-2-positive patients, but showed no significant difference with CEF and NEF regimens.HER-2 expression is not decreased after neoadjuvant chemotherapy in breast cancer patients, and HER-2-positive breast cancer can be resistant to CMF regimen, but not to CEF and NEF regimens.

To identify the differentially expressed genes associated with hypersplenism in patients with portal hypertension.The total RNA were extracted from the macrophages isolated from normal spleen and the spleen of patients with portal hypertension and reversely transcribed to cDNA with the incorporation of fluorescent (cy3 and cy5)-labeled dCTP to prepare the hybridization probes. After hybridization of Biostar-H140s chip containing 14,112 spots of cDNAs with the prepared probes, the gene chip was scanned for fluorescence intensity to screen the differently expressed genes. Three gene chips were used for hybridization and only the genes with differential expression in all the three chips were considered to associate with hypersplenism in patients with portal hypertension.Totaling 896, 1330 and 898 genes were identified to be differentially expressed by the three chips, respectively, and 121 genes (0.86%) showed differential expression in all the three chips, including 21 up-regulated known genes and 73 down-regulated known genes. The differently expressed genes were functionally related with ion channels and transport proteins, cyclins, cytoskeleton, cell receptors, cell signal transduction, metabolism, immunity, and so forth. These genes might be involved in hypersplenism in the condition of portal hypertension.cDNA microarray-based screening of differentially expressed genes in the macrophages in the spleen may provide new insights into the pathogenesis of hypersplenism in patients with portal hypertension.

To observe the change in expression of Toll-like receptor 4 (TLR4) of splenic macrophage in patients with hypersplenism due to portal hypertension (PH), and investigate the role of TLR4 of splenic macrophage in hypersplenism.Splenectomy was performed on 20 patients with hypersplenism due to PH (hypersplenism group) and 6 patients with rupture of spleen by trauma (control group) and the specimens of spleen were collected. The splenic macrophages were isolated by adhibit wall method. The expression of TLR4 of splenic macrophage was detected by immunohistochemical method (SABC), and the result was analyzed by the image analysis system. The phagocytosis of splenic macrophage was measured by chicken red blood cell (CRBC) phagocytosis assay. The level of serum endotoxin was detected before the operation by limulus assay. The results of these 2 groups were compared, and correlation analysis was made among different results of the hypersplenism group.The expression of TLR4 of splenic macrophage was 109 +/- 32 in the hypersplenism group, significantly higher than that of the control group (62 +/- 5, P0.01). The rate of phagocytosis and index of phagocytosis of splenic macrophage in the hypersplenism group were 12.6% +/- 3.0% and 0.146 +/- 0.035 respectively, both significantly higher than those of the control group (6.9% +/- 0.5% and 0.076 +/- 0.008 respectively, both P0.01) The level of endotoxin of the hypersplenism group was 0.28 EU/ml +/- 0.21 EU/ml, significantly higher than that of the control group (0.054 EU/ml +/- 0.014 EU/ml, P0.05). The rate of phagocytosis and the index of phagocytosis of splenic macrophage were notably positively correlated with the expression of TLR4 (r = 0.601, P0.01 and r = 0.553, P0.05), and the level of endotoxin (r = 0.724 P0.01 and r = 0.506, P0.05). The expression of TLR4 of splenic macrophage was positively correlated with the level of endotoxin (r = 0.525, P0.05).The expression of TLR4 of splenic macrophage in patients with hypersplenism due to PH was increased significantly. It may be one of the important factors of hypersplenism due to PH that "endotoxemia--increase of expression of TLR4 of splenic macrophage (activation of TLR of splenic macrophage)--increased destruction of red blood cells by macrophage".

Isolation of pure targeted cells is an important and essential step for the molecular analysis of tissue lesion occurred in the progression of disease. Laser capture microdissection (LCM) is a novel technique that can be applied to obtain pure targeted cell subgroup or even a single cell quickly and precisely under the microscope, thus the problem of tissue heterogeneity in molecular analysis can be tackled successfully. In this article, the principles, advantages and disadvantages of LCM were introduced. New developments in the application of LCM were summarized in two fields (DNA analysis and gene expression analysis) respectively. Meanwhile, possible directions of the future developments of LCM were put forward.