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Neutrophil extracellular trap (NET) is a type of bead-like, fibrous and reticular substances that is actively released by activated inflammatory neutrophils during the stage of infections or inflammatory responses. NET, which is composed of chromatin DNA and multiple intracellular protein components, may wrap pathogens to limit their diffusions. Meanwhile, NET may kill pathogens via a wide range of antibacterial proteins, which is considered as the third antibacterial mechanism of neutrophils, in addition to phagocytosis and degranulation. Recent studies have shown the involvement of NET in the immune response against parasitic infections. This review summarizes the advances of NETs in the immune responses against parasitic infections, so as to provide insights into the elucidation of the pathogenesis and development of therapeutics of parasitic diseases.

Background Occult peritoneal metastasis (PM) in advanced gastric cancer (AGC) patients is highly possible to be missed on computed tomography (CT) images. Patients with occult PMs are subject to late detection or even improper surgical treatment. We therefore aimed to develop a radiomic nomogram to preoperatively identify occult PMs in AGC patients. Patients and methods A total of 554 AGC patients from 4 centers were divided into 1 training, 1 internal validation, and 2 external validation cohorts. All patients’ PM status was firstly diagnosed as negative by CT, but later confirmed by laparoscopy (PM-positive n = 122, PM-negative n = 432). Radiomic signatures reflecting phenotypes of the primary tumor (RS1) and peritoneum region (RS2) were built as predictors of PM from 266 quantitative image features. Individualized nomograms of PM status incorporating RS1, RS2, or clinical factors were developed and evaluated regarding prediction ability. Results RS1, RS2, and Lauren type were significant predictors of occult PM (all P

Inflammation and fibrosis progress of nucleus pulposus (NP) cells participate in the pathologic changes of intervertebral disc degeneration (IDD). ANGPTL2 is well known for its angiogenesis and proinflammatory properties and transforming growth factor β1 (TGF-β1) is also responsible for tissue fibrosis. However, the role of ANGPTL2 in IDD and whether it is related to TGF-β1 remains unclear. This study aims to explore the relation of TGF-β1 and ANGPTL2 in the degenerative process of NP cells.We isolated NP cells of NP tissues provided from the spine fracture patients. IL-1β was used to induce the NP cells degeneration. To determine the effect of TGF-β1 and ANGPTL2 on NP cell degeneration, we regulated the cellular TGF-β1 and ANGPTL2 expression by Recombinant human protein stimulation and siRNA transfection. Quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot was employed to investigate the expression of TGF-β1, ANGPTL2, IL-6, TNF-α, collagen I, and collagen III.TGF-β1 overexpression aggravated the ANGPTL2, IL-6, TNF-α, collagen I, and collagen III expressions of NP cells that caused by IL-1β, which was rejected by ANGPTL2 gene silencing. Besides, the silencing of TGF-β1 weakened the ANGPTL2 expression. ANGPTL2 overexpression promoted the NP cells inflammation and fibrosis via increasing IL-6, TNF-α, collagen I, and collagen III expression, which was sharpened by a consequent increase of TGF-β1 expression.This study, for the first time, points that TGF-β1 aggravates degenerative NP cells inflammation and fibrosis via the mediation of ANGPTL2.

The aim of this study was to explore the changes in the body state of patients with non-small cell lung cancer (NSCLC), including intestinal flora, serum inflammatory factors, immunity and adiponectin.A total of 18 NSCLC patients (disease group) and 16 healthy people from the Medical Center (control group) were selected as research objects. The levels of immune molecules immunoglobulin A (IgA), IgG and IgM, and inflammatory factors interleukin-2 (IL-2), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and IL-6 were detected via enzyme-linked immunosorbent assay (ELISA). The level of adiponectin was determined using the quantitative kit. In addition, the changes in intestinal flora were analyzed.The overall survival time of NSCLC patients was significantly affected by IL-2 (p=0.0026), CRP (p=0.03), TNF-α (p=0.014) and IL-6 (p=0.00018). It can be seen that these inflammatory factors may play important roles in the progression of NSCLC. The levels of TNF-α (p=0.037), IL-2 (p=0.043) and CRP (p=0.000) in the peripheral blood serum were significantly higher in disease group than control group. Meanwhile, the levels of IgA (p=0.040) and IgG (p=0.000) in the peripheral blood serum were significantly higher in disease group than control group. However, no significant difference was observed in the level of IgM between the two groups (p0.05). The expression of adiponectin gene (ADIPOQ) could remarkably affect the overall survival rate of NSCLC patients, and patients with high expression of ADIPOQ exerted significantly better prognosis (p=0.017). The level of serum adiponectin was evidently higher in control group than that in disease group (p0.05). According to the linear discriminant analysis (LDA) score of the intestinal flora in both groups, the abundance of some intestinal flora (Enterobacter and Lachnospiraceae) was markedly higher in disease group than control group (p0.05). However, the abundance of Bifidobacteria, Pediococcus and Lactobacillus was remarkably higher in control group than disease group (p0.05). Correlation analysis indicated that Lactobacillus was positively correlated with Bifidobacteria (r=0.44, p=0.000), whereas was negatively correlated with Enterobacter (r=-0.22, p=0.024). Furthermore, Enterobacter was negatively associated with Bifidobacteria (r=-0.15, p=0.038) and Streptococcus (r=-0.12, p=0.046).Serum inflammatory factors, adiponectin, intestinal flora and immunity may play important roles in the development of NSCLC.

To investigate whether human umbilical cord mesenchymal stem cells (UMSCs) derived exosomes (exosome) can repair the heart after myocardial infarction (MI) by delivering circ-0001273 and its mechanism.Through the Sprague Dawley (SD) rat MI model was established, at the same time, we designed si-circ-0001273. Phosphate-buffered saline (PBS), exosome and si-circ-0001273-exosome were transplanted into ischemic hearts of rat, respectively. Through the echocardiography, hematoxylin-eosin staining (HE) method to detect the rat heart recovery. Meanwhile, H9c2 was treated with hypoxic serum-free serum to construct an in vitro apoptosis model to further explore the effect of circ-0001273 on myocardial cell apoptosis.Compared with the exosome-treated group, the left ventricular ejection fraction (EF) and shortened fraction (FS) of the rat heart was remarkably reduced and the cardiac structure was more disordered in the si-circ-0001273-exosome-treated group. Meanwhile, in vitro TUNEL staining and flow cytometry detection, results showed that compared with the exosome co-culture group, the incidence of H9C2 cell apoptosis in the si-circ-0001273-exosome co-culture group was obviously increased.Circ-0001273 can remarkably inhibit the occurrence of myocardial cell apoptosis in ischemic environment, promote MI repair, and provide a good reference for clinical treatment.

In 2016 WHO classification, EBV +DLBCL of the elderly was replaced by EBV+ DLBCL NOS. This is due to the fact that many young patients of EBV+ DLBCL were found in recent years.In this study, we retrospectively analyzed clinical features and survival outcomes of EBV positive DLBCL patients in different age groups. All the patients treated at a single center.When we use different ages (40, 50 and 60 years old) as cutoffs, the prevalence of EBV positive DLBCL was 12.0%, 12.3% and 13.0% in younger patients and 19.0%, 15.4% and 13.8% in elder patients respectively. Whatever the age cutoff was, EBV positive associated with unfavorable clinical prognosis in elder groups. When we use 40 and 50 years old as age cutoffs, poor impacts of EBV positive on overall survival and progression-free survival were observed only in elder patients, but not in younger patients. It should be noted that when we use 60 years old as age cutoff, the results were the opposite.EBV+ DLBCL patients with age of 40 to 60 years old showed poorer prognostic features than EBV- DLBCL patients; however, patients in other age groups did not show evident differences in prognosis between EBV+ DLBCL patients and EBV- DLBCL patients. This finding was not reported before.

The aim of this study was to investigate the effect of leptin (Lep) on the proliferation, invasion and apoptosis of prostate cancer cells through the extracellular regulated protein kinase 1/2 (ERK1/2) signaling pathway.Prostate cancer DU145 cells in the logarithmic growth phase were randomly divided into Lep (10, 20, 40, 80, 160 and 320 ng/mL) groups and blank control (Con) group. After culture, the cells were treated for 6 h, 12 h and 24 h, respectively. The effects of Lep on the proliferation and invasion of DU145 cells were detected via methyl thiazolyl tetrazolium (MTT) assay and transwell chamber assay, respectively. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was carried out to examine the messenger ribonucleic acid (mRNA) expressions of ERK1/2, b-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) in DU145 cells after Lep treatment for 24 h. Thereafter, immunofluorescence assay was performed to detect the localization of ERK1/2 protein in prostate cancer DU145 cells. In addition, the expressions of phosphorylated (p)-ERK, ERK1/2 and apoptosis-related proteins, Bcl-2, Bax and cleaved cysteinyl aspartate specific proteinase (c-Caspase 3) in prostate cancer DU145 cells after treatment with different concentrations of Lep for 24 h were examined by Western blotting.MTT assay results showed that the proliferation rate of DU145 cells increased significantly at 6 h, 12 h and 24 h after 5-320 ng/mL of Lep treatment (p0.05). Transwell assay manifested that the number of invasive cells was significantly raised after Lep treatment for 24 h (p0.05). Meanwhile, the invasion ability of cells increased gradually with the elevation of Lep concentration. Subsequent qRT-PCR results demonstrated that after treatment with different concentrations of Lep, the mRNA expressions of ERK1/2 and Bcl-2 rose markedly (p0.05). However, the mRNA expression of Bax was remarkably down-regulated (p0.05) with the increase of Lep concentration in a concentration-dependent manner. According to the detection using a laser scanning confocal microscope, ERK1/2 red fluorescence showed punctiform aggregation, which was gradually raised with the increase of Lep concentration for 24 h. Moreover, Western blotting results denoted that with the increase of Lep concentration, the protein expressions of p-ERK, ERK1/2 and Bcl-2 were notably elevated (p0.05), while those of Bax and c-Caspase 3 were distinctly reduced (p0.05).Lep activation induces the proliferation, promotes the invasion and inhibits the apoptosis of prostate cancer cells through the ERK1/2 signaling pathway.

The aim of this study was to explore the effect of micro ribonucleic acid (miR)-200c on the proliferation and apoptosis of Wilms tumor cells, and to further elucidate its potential mechanisms.Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was used to detect the expression level of miR-200c in cancer tissues and adjacent normal tissues of 20 patients with Wilms tumor. Human primary Wilms tumor cells were taken as research objects, and were divided into Control group and miR-200c mimic group. In miR-200c mimic group, miR-200c was overexpressed in Wilms tumor cells using liposome transfection technology. Subsequently, the proliferation and apoptosis of cells in each group were observed by functional assays. The expression levels of phosphorylated protein kinase B (p-Akt), total Akt (T-Akt) and glucose transporter protein type 1 (GLUT1) in each group of cells were finally detected by Western blotting.In Wilms tumor patients, the expression level of miR-200c in cancer tissues was notably lower than that in adjacent normal tissues (p0.05). Wilms tumor cells were cultured in vitro, and were transfected with miR-200c mimic. Subsequent cell counting kit-8 (CCK-8) assay results showed that the proliferation ability of cells in miR-200c mimic group was remarkably weakened (p0.05). Colony formation assay indicated the number of formed colonies in miR-200c mimic group was remarkably less than that in Control group (p0.05). Western blotting results manifested that overexpression of miR-200c markedly increased the ratio of B-cell lymphoma protein 2 (Bcl-2) to Bcl-2-associated X protein (Bax) (p0.05). Flow cytometry results revealed that miR-200c overexpression significantly elevated the apoptosis rate of Wilms tumor cells (p0.05). In addition, it was discovered that the overexpression of miR-200c could prominently reduce the phosphorylation level of intracellular Akt and the expression of its downstream protein GLUT1. Finally, immunohistochemical staining results verified that the expression levels of p-Akt and GLUT1 in cancer tissues of Wilms tumor patients were significantly higher than those in adjacent normal tissues (p0.05).MiR-200c was lowly expressed in cancer tissues of Wilms tumor patients. Besides, overexpression of miR-200c inhibited the proliferation and promoted the apoptosis of cells through targeted inhibition of the Akt/GLUT1 signaling pathway.

We aimed at studying the correlation between TGIF2 expression and clinicopathological features of cervical cancer (CCa). The relationship between TGIF2 and FCMR and its influence on the proliferation and metastasis of tumor cells were investigated using molecular biology techniques, so as to reveal the pathogenesis of CCa and provide a new target for clinical treatment.TGIF2 expression in 60 pairs of cervical tumors and paracancerous tissues samples collected from CCa patients of our hospital was studied by quantitative real-time polymerase chain reaction (qPCR) analysis, and the association between TGIF2 expression and the clinical indicators or prognosis of CCa patients were analyzed. CCa cells with TGIF2 knockdown were constructed using transfection technology. Changes in the biological phenotypes (proliferation, migration, invasion) of CCa cells C33-A and HeLa after TGIF2 knockdown were determined by Cell Counting Kit-8 (CCK-8) and transwell assays. In addition, the effects of TGIF2/FCMR axis on CCa metastasis were further explored in nude mice in vivo.Our data revealed a significant increase in TGIF2 mRNA expression in CCa tissue specimens compared to adjacent ones, and the increasing degree was positively correlated with the incidence of lymph node or distant metastasis of CCa patients. The results of CCK-8 and transwell suggested that knocking down TGIF2 effectively attenuated the proliferative ability and invasiveness of CCa cells. Luciferase assay confirmed that TGIF2 can directly bind to the DNA promoter of its target gene FCMR. Simultaneous transfection of sh-TGIF2 and sh-FCMR partially reversed the inhibitory effect of single transfection of TGIF2 knockdown on the malignant progression of CCa. Experiments in nude mice also suggested that TGIF2 could promote CCa tumorigenesis through the modulation of FCMR expression.In summary, TGIF2 can promote the migration and proliferation ability of cervical cancer cells via down-regulating FCMR. Our study provides a new therapeutic target for the clinical treatment of cervical cancer.

OBJECTIVE: To explore the correlation between different personality characteristics and self-management attitude such as medication, exercise and diet in patients with type 2 diabetes mellitus. METHODS: The patients with type 2 diabetes mellitus from 4 Community Healthcare Services and 22 affiliated community stations of Tongzhou District and Shunyi District of Beijing were selected as subjects. The Chinese big five personality inventory and the self-designed scale including the attitudes of medication, exercise and diet were used in the study. RESULTS: In this study, 642 subjects were finally included, among whom the sex ratio of male and female was basically the same, 61.21% were over 61 years old. In this study, different genders had differences in neurotic personality (P

To investigate the advantages of thoracoabdominal radical gastrectomy for advanced Siewert type Ⅱ adenocarcinoma of the esophagogastric junction.Clinical data of 86 patients with Siewert type Ⅱ adennocarcinoma of the esophagogastric junction who received surgical treatment at the Henan Provincial Tumor Hospital from January 2015 to January 2016 were retrospectively analyzed. Among them, 44 patients underwent abdominal operation (abdominal group), and 42 patients underwent thoracoabdominal radical gastrectomy (thoraco-abdominal group). The operation time, lymph node number, distance between the tumor and cutting edge, amount of intraoperational blood loss, postoperative pulmonary complications, and postoperative hospital stay in the two groups were compared.Comparing the thoracoabdominal group with the abdominal group, the number of removed lymph nodes was 41.57±9.22 vs. 35.09±10.61 (P0.01), the number of removed mediastinal lymph nodes was 6.38±1.50 vs. 3.52±1.42 (P0.01), the distance between the tumor and cut edge was (5.62±0.73) cm vs. (3.30±0.85) cm (P0.01), whereas the operation time, intraoperative blood loss, postoperative pulmonary complications, occurrence of anastomotic leakage and hospital stay were statistically not significantly different (P0.05 for all).For patients with advanced Siewert type Ⅱ adenocarcinoma of the esophagogastric junction, radical gastrectomy through thoracoabdominal approach can resect a longer segment of the esophagus, dissect more mediastinal lymph nodes, and does not increase post-operative complications and extend hospital stay, thus, exhibits obvious advantages in the surgical treatment of Siewert Ⅱ adenocarcinoma of the esophagogastric junction.

The correlation of vitamin D receptor (VDR) gene polymorphism and blood lead level had been in doult, which allowed us to write this article.Relevant studies about the blood lead and VDR B/b gene polymorphism which published from 1979-2015, were searched in multiple bibliographic databases, such as: CNKI, Wanfang Database, PUBMED. Of the ten references selceted, three were divided into two groups which were classified as different researches, so there were thirteen studies in the meta-analysis. According to the level of blood lead, the studies were analyzed by three groups: normal group, low dose grou and high dose group. The analysis was performed by stata 12.0 software.The blood lead level of VDR B/b genotype was significantly difference in all the three groups (P0.05) , but there were apparent heterogeneity between normal group and low dose group (P0.05, I(2)=84.2%; P0.05, I(2)=88.9%) , except the high dose group (P0.05, I(2)=12.7%) ; after adjusted, all showed no heterogeneity, and the results were still the same.The genotype of VDR may be correlated with blood lead, and the levels of blood lead varied with different genetypes.

Polycomb protein histone methyltransferase enhancer of Zeste homologe 2 (EZH2) is frequently overexpressed in human malignancy and is implicated in cancer cell proliferation and invasion. However, it is largely unknown whether EZH2 has a role in modulating DNA damage response. Here, we show that EZH2 is an important determinant of cell fate decision in response to genotoxic stress. EZH2 depletion results in abrogation of both cell cycle G1 and G2/M checkpoints, directing DNA damage response toward predominant apoptosis in both p53-proficient and p53-deficient cancer cells, but not in normal cells. Mechanistically, EZH2 regulates DNA damage response in p53 wild-type cells mainly through transcriptional repression of FBXO32, which binds to and directs p21 for proteasome-mediated degradation, whereas it affects p53-deficient cells through regulating Chk1 activation by a distinct mechanism. Furthermore, pharmacological depletion of EZH2 phenocopies the effects of EZH2 knockdown on cell cycle checkpoints and apoptosis. These data unravel a crucial role of EZH2 in determining the cancer cell outcome following DNA damage and suggest that therapeutic targeting oncogenic EZH2 might serve as a strategy for improving conventional chemotherapy in a given malignancy.

Deregulation of the pRB/E2F pathway, which occurs frequently in human malignancy, is often associated with inappropriate proliferation and/or apoptosis. While the role of E2F1 in apoptosis induction has been well-established, it remains unclear how this pro-apoptotic activity is regulated in cancer. Here we describe EZH2, an oncogenic polycomb histone methyltransferase and an E2F1 target, as an important regulator of E2F1-dependent apoptosis. We show that E2F1 induces EZH2 expression, which in turn antagonizes the induction of E2F1 pro-apoptotic target Bim expression. RNAi-mediated gene depletion of EZH2 enhances E2F1-dependent Bim expression, thereby promoting the pro-apoptotic activity of E2F1. Hence, the concomitant induction of EZH2 and Bim by E2F1 constitutes a fail-safe mechanism to allow tumor cells with aberrant E2F1 activity to evade apoptosis. These findings reveal a novel mechanism by which the apoptotic activity of E2F1 is restrained in human cancer and also provide the first evidence that EZH2 directly regulates apoptotic process in cancer cells.

Massage therapy is one of the most widely accepted alternative form of medicine helping patients suffering from varied pathological states including arthritis, anxiety, sleep problems, pain management and injury repair. Besides this, it is one of the safest forms of alternative medicine and has become favorite among various health care professionals. However, there is still a lot of debate is going in medical world pertaining to its certain use in modern medicine. So, the present review shall enlighten all the latest aspects of massage therapy in current medicine.

Breast cancer remains a major health problem even with all the recent technological advancements. Large-scale gene expression analysis has offered great ease for both biological characterization and therapeutic planning of breast cancer. Previous studies mostly used variance/regression analysis, which becomes fundamentally flawed when there are unaccounted array specific factors. Here we aim to investigate the underlying mechanism of breast cancer through partial least squares (PLS) based gene expression profile analysis.With a gene expression profile data set downloaded from the Gene Expression Omnibus database, we performed PLS based analysis.We acquire 932 and 771 differentially expressed genes (DEGs) in breast cancer metastasis of estrogen-receptor (ER)-positive and ER-negative patients, respectively. For ER-positive patients, 32 pathways were found to be enriched with DEGs, including immune related pathways, cellular processes and environmental information processing pathways. Survival analysis demonstrated that 18 of them were closely related with non-recurrence rate along time after surgery. For ER negative patients, only three pathways including the folate biosynthesis pathway were enriched with DEGs and none of them overlapped with those of ER positive patients. Only the cholinergic synapse pathway was significantly associated with the non-recurrence rate according to the survival analysis.Our findings shed light on pathways involved in breast cancer relapse with the hope to give some theoretical supports for further therapeutic study.

Direct PCR detection of bacteria in clinical samples is often hindered by the presence of compounds that inhibit the PCR. To improve and accelerate the diagnosis of Mycobacterium avium-M. intracellulare complex infections, an immunomagnetic PCR (IM-PCR) assay was developed. This IM-PCR procedure combines the separation of mycobacteria by antimycobacterial monoclonal antibody coupled to magnetic beads with an M. avium-M. intracellulare complex-specific PCR protocol based on 16S rRNA gene sequences. As few as 10 M. avium bacilli were detected in spiked human stool samples, a clinical specimen usually refractory to conventional PCR analysis, by the IM-PCR method. Moreover, M. avium organisms were detected in about 24 h in 18 of 22 culture-confirmed fecal samples from AIDS patients. This IM-PCR protocol should allow for the rapid and sensitive detection of M. avium isolates in clinical specimens.

A shared-primer PCR method for the detection of infection was developed by using primers derived from DNA sequences upstream of the structural genes for the porin proteins of Bordetella pertussis and Bordetella parapertussis. This method resulted in a 159-bp PCR product specific for B. pertussis and a 121-bp DNA fragment specific for B. parapertussis and allowed for the simultaneous detection of these pathogens. The PCR procedure was shown to be very specific since no PCR product was obtained from 36 non-Bordetella bacterial DNAs. Nasopharyngeal aspirates (NPAs) from children suspected of having pertussis were evaluated by the PCR method, culture, and the Chinese hamster ovary (CHO) cell assay, which detects pertussis toxin. B. pertussis was cultured from 119 of 205 NPAs assayed, and the presence of pertussis toxin was detected in 69 of the NPAs by the CHO cell assay. When ethidium bromide staining was used to detect PCR products, 100 NPAs gave positive results by shared-primer PCR; 94 of these NPAs were also positive by culture. The result indicated a sensitivity of 79% for PCR when culture was used as the standard. The sensitivity of PCR was increased to 95% when a digoxigenin immunoblot system was used. An additional 20 NPAs from patients with suspected pertussis that were culture negative also gave positive results by PCR. The specific and sensitive PCR method described here should be useful for both the clinical diagnosis of pertussis and case identification in vaccine trials.

To evaluate antibody responses to a panel of four protein antigens secreted from Mycobacterium tuberculosis, CFP-21, ESAT-6, MPT-63 and MPT-64, and their complex antigen.Seventy-eight samples of serum and cerebral spinal fluid were tested by ELISA.Serum IgM antibody positive rates of protein antigens CFP-21, ESAT-6, MPT-63 and MPT-64 and their multi-antigen complex were respectively 34.6%, 60.3%, 52.6%, 78.2% and 96.2%; IgG antibody positive rates were respectively 46.2%, 64.1%, 93.6%, 57.7% and 84.6%. The specificity of the antibodies to the four proteins secreted from M. tuberculosis and its mixture antigen was 100%; sensitivity to the multi-antigen complex was higher than for the four single antigens. Among the IgM positive rates, the differences between the multi-antigen complex and CFP-21, ESAT-6, MPT-63 and MPT-64 were significant (P0.01); among the IgG positive rates, the differences between the multi-antigen complex and CFP-21, ESAT-6 and MPT-64 were significant (P0.01). Although the IgG positive rate for MPT-63 was higher than for the multi-antigen complex, the difference was not significantly different (P0.05).These findings indicate that the multi-antigen complex composed of CFP-21, ESAT-6, MPT-63 and MPT-64 may be useful in the diagnosis of TB.

The immunogenicity of most polysaccharides (PSs) contained in the pneumococcal vaccine is low in children less than 2 years of age. Enhancement of immune response in early life can be induced by immunization of pneumococcal glycoconjugate as well as plasma DNA coding for cell-surface protein antigen. Pneumococcal type 19F PS conjugated with inactivated pneumolysin (Ply) induced in mice remarkable antibody responses to both type 19F PS and the protein carrier. In addition, the conjugate was administered to pregnant mice during gestation and/or lactation, and to their offspring during early infancy. When the young mice were challenged with type 19F pneumococci, the bacteria were cleared more rapidly from the blood of immunized mice than from that of the control group. The mortality rate of young mice from immunized mothers was also significantly lower than the control group. These results indicate that the effective protective immunity against pneumococcal infection can be induced in young mice by the maternal immunization with the glycoconjugate during gestation and at early infancy. Studies have been conducted to express type 19A pneumolysin gene (ply) in mammalian cells. Ply DNA was inserted into the cloning site of a vector containing CMV promoter. The recombinant plasmid DNA containing ply was transfected in human rhabdomyosarcoma cells and the gene expression was confirmed by immunoblot. Injection of mice three times 50-100 ug per dose ply DNA produced high serum levels of Ply IgG and IgM antibodies and showed rapid bacterial clearance from the blood.

To investigate the mechanism of apoptosis of HL60 cells induced by the annonaceous acetogenin, squamocin.Induction of apoptosis was determined through Hoechst33258 dye staining and DNA agarose gel electrophoresis. Expression of the proteins was detected using Western blot analysis. Caspase-3 activity was detected using caspase-3 kit.Treatment of HL-60 cells with squamocin resulted in extensive nuclear condensation, DNA fragmentation, cleavage of the death substrate poly(ADP-ribose) polymerase (PARP) and induction of caspase-3 activity. Pretreatment of HL-60 cells with caspase-3 specific inhibitor DEVD-CHO prevented squamocin-induced DNA fragmentation, PARP cleavage and cell death. Stress-activated protein kinase (SAPK/JNK) was activated after treatment with squamocin in HL-60 cells.These results suggest that apoptosis of HL-60 cells induced by squamocin require caspase-3 activation, and could be related to SAPK activation.

To study the biological function of ceramide signaling in Bel7402 cells.Inhibition of cell growth was assayed using MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was detected by electrophoresis and flow cytometry. The levels of protein p53, Bcl-2, and Bax were measured with Western blot.Bel7402 cells treated with C2-ceramide underwent cell proliferation inhibition. IC50 value was 14.28 mumol.L-1. After treatment of Bel7402 with ceramide, the morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed. SubG1 peaks were detected on flow cytometry (FCM). Agarose gel electrophoresis of DNA from cells treated with ceramide revealed "ladder" pattern. The Western blot assay from cell extracts showed that the levels of protein p53 were decreased after ceramide treatment. The levels of protein Bcl-2 were decreased also. But the levels of Bax protein showed no difference between untreated cells and treated cells.Ceramide induces apoptosis in Bel7402 cells, related to Bcl-2 down-regulation.

The leukocyte CD44 and CD45 cell surface receptors are associated via the linker proteins ankyrin and fodrin with the cytoskeleton, which itself is important in immune cell functions such as adherence, chemotaxis, and phagocytosis. The effects of rat antihuman CD44 and CD45 monoclonal antibodies on phagocytosis of fluoresceinated heat-killed Staphylococcus aureus 502A by normal human neutrophils (PMNs) during 2 hr incubation in RPMI-1640 was studied via flow cytometry and confocal microscopy. Flow cytometry was performed using an excitation wavelength of 488 nm, fluorescence being measured at 515-560 nm on 50,000 PMNs per sample. Confocal microscopy was performed on samples after further incubation with rhodamine-conjugated antiankyrin. Anti-CD44 resulted in an increase of 27-31% compared to control (P = 0.004) in the proportion of PMNs fluorescing, an increase of 17-24% (P = 0.001) in mean intracellular fluorescence per PMN, and an increase in total PMN fluorescence of 50-58% compared to control (P0.001). In contrast, anti-CD45 had little effect on phagocytosis. Colchicine (a microtubule-disrupting agent) enhanced, whereas cytochalasin-D (a microfilament inhibitor) inhibited bacterial phagocytosis; cytochalasin-D completely abrogated the effect of anti-CD44 on this PMN function. Hyaluronic acid augmented phagocytosis by an increment similar to that observed with anti-CD44. Two-color flow cytometry and confocal microscopy demonstrated that ankyrin always colocalized with ingested fluorescein isothiocyanate (FITC)-labeled bacteria. These data strongly suggest that CD44 is involved in bacterial phagocytosis, provide further evidence of CD44 receptor linkage to cytoskeletal elements in human leukocytes, and suggest that ankyrin has a significant role in the transport of phagosomes.

That L-arginine (L-Arg) augments the host response to acute bacterial sepsis suggests that this amino acid intervenes early in the immune response, perhaps via the nitric oxide synthetase (NOS) pathway. The effect of L-Arg supplementation on in vitro phagocytosis of fluorescein-labeled, heat-killed Staphylococcus aureus by peripheral blood neutrophils (PMNs) from 12 normal human volunteers was studied. Separated PMNs were incubated for 2 h with labeled bacteria, with and without supplemental L-Arg, D-arginine, glycine, and/or the NOS inhibitors L-canavanine, aminoguanidine, or L-NG-nitroarginine methyl ester. PMNs were fixed and extracellular fluorescence quenched with crystal violet. By flow cytometry and confocal microscopy, L-Arg supplementation was shown to result in a highly significant increase in PMN bacterial phagocytosis, the maximal effect being seen with L-Arg 380 microM and falling off with higher concentrations. This augmentation was completely abrogated by NOS inhibitors in molar excess, but inhibitors alone did not suppress phagocytosis below that of unsupplemented controls. Neither D-arginine nor glycine affected phagocytosis; the L-Arg effect was stereospecific and not related to utilization of L-Arg as an energy source. L-Arg supplementation significantly enhances bacterial phagocytosis in human neutrophils, perhaps by effects on cytoskeletal phenomena, and this appears to be mediated through NOS activity. Phagocytosis by nonspecific immune cells which intervene early in the response to sepsis is critically important, and beneficial effects of L-Arg on the clinical course of sepsis may be due at least in part to augmentation of phagocyte function.

Asthma is one of the most frequent and common diseases in China. It seriously threatens the health of the population. It is evident that mites present in rural houses may serve as an allergic antigen. In our survey, we have found several kinds of mites in farmers' houses in the northeastern part of China which have very close relation with asthmatic diseases. Investigations in rural houses further proved that the cause of asthma is certainly related with the allergic antigen of mites. The methods of prevention and control of mites are enumerated.

Endomyocardial biopsy (EMB) was performed in 24 cases suspected of having myocarditis and 42 cases with cardiac symptoms and signs of unknown etiology. There was an obvious correlation between the clinical manifestations and the pathological findings in cases suspected of having myocarditis (r = 0.5296, P less than 0.005). The ratio of correspondence was 78.8%. It is shown that EMB is more sensitive than 201TI myocardial imaging in diagnosing myocarditis and serial EMB can follow its clinical process closely. Moreover, EMB can discriminate myocarditis from myocardiopathy.

An in vitro study by using cross-linked agarose coated activated charcoal (CAAC-II) as adsorbent was conducted. Endotoxin was quantitatively measured by chromogenic method of limulus lysate test. Results of the study demonstrated that endotoxin was fairly efficiently removed by CAAC-II. It may therefore be possible for effective adsorption of endotoxin in vivo studies and warrant further experimental study for CAAC-II hemoperfusion in clinical endotoxemia.

Various types of macroporous resin beads were screened for the adsorption of middle molecules in vitro. In order to obtain high adsorption capacity, the pore size of the resin should be large enough to allow diffusion of the middle molecules. The adsorption capacity of X-5 resin for Vitamin B12, inulin and cytochrome C was 35.3 mg/g, 1.2 mg/g and 42.9 mg/g respectively in vitro tests. Hemoperfusion with acute renal failure rat model showed good clearance (0.35 +/- 0.25) with X-5 resin for middle molecules.