[The Breast Cancer Cohort Study in Chinese Women: research design and preliminary results of clinical multi-center cohort].

Objective: The incidence of breast cancer in Chinese women continues to rise. The large breast cancer cohort studies in China are relatively scarce. There are many bottlenecks in the construction of large clinical cohort for breast cancer diagnosis, treatment, and prognoses, such as inconsistent standards, high rates of lost follow-up, repeated construction, and inability to share. To better solving the difficulties and problems faced by large-scale clinical cohort research in China, this project will cooperate with several tertiary A hospitals to establish a breast cancer cohort in Chinese women. It also provides a data platform and technical support for breast cancer multi-center clinical cohort research. Methods: Based on the evidence-based medicine and expert opinion and consensus, we established a breast cancer cohort standardized indicator set-recording baseline information, diagnosis and treatment-related information of the enrolled patients, and collecting biological specimens. According to the technical specification of long-term follow-up for the endpoint, data management, and data security and in the large population-based cohort study, a standardized follow-up system for the diagnosis, treatment, and prognosis of breast cancer prospective cohorts is formed. Results: Based on standardized data sets and the computer discipline's advantage from the University of Science and Technology Beijing, we integrate the new information technology methods, including dynamic information collection terminals and social networks. Thus, the quality of control programs on compliance and intelligence data was improved, and a Chinese women breast cancer cohort database was developed. By February 2020, 12 147 patients were included in the clinical cohort database. Biological specimens'resources in cohort construction were collected and cooperated with Shandong University to research the multi-center quality control system and shared evaluation system of biobanks. Building an open and shared biobank network and forming a full chain of breast cancer research platform. Conclusion: With the implementation of the "13(th) Five-Year Plan" precision medicine research, this study provides a research foundation for precision diagnosis and treatment of breast cancer and provides data support for the country to formulate relevant medical policies.

Resveratrol alleviates osteoporosis through improving the osteogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE: To investigate the protective effect of Resveratrol (RES) on TNF-α-induced inhibition of osteogenic differentiation, thus alleviating the progression of osteoporosis (OP). MATERIALS AND METHODS: OP model in rats was first conducted by performing ovariectomy (OVX). Rats were randomly divided into sham group, OVX group, and RES+OVX group. Body weight of each rat was regularly recorded every week. Bone mineral density (BMD) of rat femoral metaphysis was measured by micro-CT. Changes in radial degrees and loads of rat femora were examined through three-point bending experiments. Relative levels of OCN and Runx2 in each group were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Alkaline phosphatase (ALP) activity and calcification ability were assessed through ALP staining and alizarin red staining, respectively. Bone mesenchymal stem cells (BMSCs) were extracted from healthy rats and divided into control group, Tumor necrosis factor-α (TNF-α) group, RES group, and TNF-α+RES group based on different treatments. Relative levels of OCN and Runx2, ALP activity, and calcification ability in each group were detected in the same way. Finally, protein levels of NF-κB and ß-catenin in BMSCs were determined. RESULTS: Rats in each group gained body weight during the experimental period, especially those in OVX group and RES+OVX group. No significant difference in the body weight was found between OVX group and RES+OVX group. BMD in rat femora of RES+OVX group was higher than in OVX group but lower than sham group. Elastic/max radial degree and elastic/max load of femora were markedly reduced in OVX group compared to RES+OVX group. Relative levels of OCN and Runx2, ALP activity and calcification ability decreased in OVX group relative to sham group, which were partially reversed by RES treatment. After osteogenic differentiation in BMSCs induced with TNF-α, viability and calcification ability were markedly reduced and were upregulated by RES treatment. Moreover, RES treatment enhanced the downregulated levels of OCN and Runx2 in BMSCs undergoing TNF-α induction. Upregulated protein levels of nuclear factor kappa-B (NF-κB) and ß-catenin in TNF-α-induced BMSCs were downregulated by RES treatment. CONCLUSIONS: The inhibited osteogenic differentiation of BMSCs undergoing TNF-α induction is improved by resveratrol treatment, which contributes to alleviate the progression of osteoporosis.

Effects of miR-26a/miR-146a/miR-31 on airway inflammation of asthma mice and asthma children.

OBJECTIVE: This study detected the expressions of microRNA-26a (miR-26a), miR-146a and miR-31 in lung tissues and BALF (bronchoalveolar lavage fluid) of asthma mice and children. Besides, cytokine levels of interleukin-5 (IL-5), IL-8, IL-12 and tumor necrosis factor-α (TNF-α) were detected as well. We aim to provide an experimental basis for clinical treatment of asthma. PATIENTS AND METHODS: Forty female BALB/c mice were randomly assigned into control group and asthma group, respectively. Mice in asthma group (n=20) were immunized by intraperitoneal injection of OVA (ovalbumin) and provoked by atomization inhalation of OVA from the 15th day for 10 days. Mice in control group (n=20) were immunized and provoked with isodose saline during the same period. At the 26th day, mice were sacrificed for collecting lung tissues and BALF. Besides, we enrolled 17 cases of asthma children and 13 cases of children with airway foreign body as controls. BALF of each subject was collected. Total cellular score and differential counting in BALF were recorded. Expression levels of miR-26a, miR-146a, and miR-31 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Levels of IL-5, IL-8, IL-12, and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: The total cellular score in BALF of asthma mice and asthma children was higher than that of controls (p<0.05). Percentages of eosinophils, neutrophils, and lymphocytes in BALF of asthma mice and asthma children were higher than those of controls, whereas the percentage of macrophages was lower (p<0.05). Levels of IL-5, IL-8, IL-12, and TNF-α in lung tissues of asthma mice were markedly elevated compared with those of controls (p<0.05). Similarly, levels of IL-5, IL-8, IL-12, and TNF-α were higher in BALF of asthma children than controls (p<0.05). RT-PCR data showed higher mRNA levels of miR-26a, miR-146a, and miR-31 in lung tissues of asthma mice than controls (p<0.05). The mRNA levels of miR-26a, miR-146a, and miR-31 in BALF of asthma children were highly expressed compared with those of controls as well (p<0.05). CONCLUSIONS: MiR-26a, miR-146a, and miR-31 are involved in asthma progression mainly through regulating inflammatory factors and cells.

Effects of isobutyrate supplementation on ruminal microflora, rumen enzyme activities and methane emissions in Simmental steers.

The objective of this study was to evaluate the effects of isobutyrate supplementation on rumen microflora, enzyme activities and methane emissions in Simmental steers consuming a corn stover-based diet. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. The treatments were control (without isobutyrate), low isobutyrate (LIB), moderate isobutyrate (MIB) and high isobutyrate (HIB) with 8.4, 16.8 and 25.2 g isobutyrate per steer per day respectively. Isobutyrate was hand-mixed into the concentrate portion. Diet consisted of 60% corn stover and 40% concentrate [dry matter (DM) basis]. Dry matter intake (averaged 9 kg/day) was restricted to a maximum of 90% of ad libitum intake. Population of total bacteria, cellulolytic bacteria and anaerobic fungi were linearly increased, whereas that of protozoa and total methanogens was linearly reduced with increasing isobutyrate supplementation. Real-time PCR quantification of population of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Fibrobacter succinogenes was linearly increased with increasing isobutyrate supplementation. Activities of carboxymethyl cellulase, xylanase and ß-glucosidase were linearly increased, whereas that of protease was linearly reduced. Methane production was linearly decreased with increasing isobutyrate supplementation. Effective degradabilities of cellulose and hemicellulose of corn stover were linearly increased, whereas that of crude protein in diet was linearly decreased with increasing isobutyrate supplementation. The present results indicate that isobutyrate supplemented improved microflora, rumen enzyme activities and methane emissions in steers. It was suggested that the isobutyrate stimulated the digestive micro-organisms or enzymes in a dose-dependent manner. In the experimental conditions of this trial, the optimum isobutyrate dose was approximately 16.8 g isobutyrate per steer per day.

Expression of vimentin and survivin in clear cell renal cell carcinoma and correlation with p53.

PURPOSE: This study focuses on investigating the expression correlation of vimentin, survivin and p53 in clear cell renal cell carcinoma (ccRCC) and the clinical significance. METHODS: The mRNA and protein expression levels of the vimentin, survivin and p53 were determined in ccRCC and adjacent normal renal tissues, using quantitative real-time-polymerase chain reaction (qRT-PCR) and Western blot. We detected the expression and localization of vimentin, survivin and p53 protein in ccRCC by immunohistochemistrical SP method and analyzed the relationships among clinical pathologic parameters and patient prognosis. RESULTS: The expression of vimentin and survivin was significantly increased in ccRCC compared with adjacent normal renal tissues, which were positively correlated with the pathological grade and clinical stage (P < 0.05). p53 was highly expressed in ccRCC compared with normal tissues (P < 0.05), which was not positively correlated with the pathological grade and clinical stage (P > 0.05). Furthermore, univariate and multivariate analysis showed that high expression levels of vimentin and survivin were independent prognostic indicators for ccRCC. The levels of vimentin and survivin were positively correlated in ccRCC (r = 0.428, P < 0.01). CONCLUSIONS: Reliable basis about biological behavior and prognosis judgments of ccRCC can be provided by combining detection of vimentin and survivin. Foundation and new ideas for gene therapy of ccRCC may be provided by further studying the relationship among vimentin, survivin and p53 in ccRCC.

Preparation and evaluation of zirconia-coated silica monolith for capillary electrochromatography.

Silica monoliths were fabricated inside fused-silica capillaries. Then the monolithic columns were coated with membrane-like zirconia. The zirconia-coated silica monoliths exhibited different EOF behavior comparing with that of bare silica monoliths. The magnitude and direction could be manipulated by changing the running buffers. Due to the amphoteric characteristic of zirconia, the silica monoliths with zirconia surface facilitate the separation of basic compounds. Aromatic amines and alkaloids were separated without obvious peak tailing. The zirconia surface was easily modified with octadecylphosphonic acid for the separation of neutral compounds. Column efficiency as high as 90,000 and 80,000m(-1) was obtained for beberine and naphthalene, respectively. Furthermore, the zirconia coating increased the stability of the monolithic columns. Even after being exposed to severe condition, there was no apparently efficiency decrease for the test samples.

Effect of recombinant bactericidal/permeability-increasing protein on endotoxin translocation and lipopolysaccharide-binding protein/CD14 expression in rats after thermal injury.

OBJECTIVE: To investigate the potential mechanisms underlying the in vivo effect of recombinant bactericidal/permeability-increasing protein (rBPI21) on endogenous bacteria or endotoxin translocation and lipopolysaccharide-binding protein/CD14 expression secondary to thermal injury. DESIGN: Prospective, randomized, controlled animal study. SETTING: College hospital animal research laboratory. SUBJECTS: Thirty-six male Wistar rats weighing 250-300 g. INTERVENTIONS: The rats were anesthetized, and a 35% total body surface area full-thickness burn was created. Animals were randomized to receive treatment with either rBPI21 or the control protein (albumin). rBPI21 (2 mg/kg body wt, BPI group) or a protein control preparation (burn group) in the same dose was administered in an intravenous bolus at 30 mins and 4 hrs after thermal injury. All animals were killed at 12 and 24 hrs postburn (six to ten rats for each interval). In addition, eight rats were taken as normal controls. MEASUREMENT AND MAIN RESULTS: Our data showed that treatment with rBPI21 was effective in preventing endotoxin translocation secondary to severe burns. Meanwhile, tissue lipopolysaccharide-binding protein, CD14, and tumor necrosis factor-alpha mRNA expression in various organs were inhibited markedly by rBPI21 secondary to acute insults (p <.05-.01). Furthermore, significant reduction in serum aminoleucine transferase concentrations and elevation in intestinal diamine oxidase activities in the rBPI21-treated group were found compared with controls (p <.05-.01). CONCLUSIONS: These findings indicate that endotoxin accumulated in local sites after thermal injury can markedly up-regulate lipopolysaccharide-binding protein/CD14 and tumor necrosis factor-alpha mRNA expression in various organs. Meanwhile, up-regulation of lipopolysaccharide-binding protein/CD14 expression would be the major molecular mechanism of increasing sensitivity to endogenous endotoxin response after burns. Early treatment with rBPI21may be effective in attenuating multiple organ damage resulting from gut-origin endotoxin translocation. This might be associated with the down-regulation effects of tissue lipopolysaccharide-binding protein and CD14 gene expression by the use of rBPI21.

[The biological effect of tetrahydrobiopterin and its potential role in sepsis].

It is well known that overproduction of nitric oxide (NO) is the final common pathway in septic shock. Tetrahydrobiopterin (BH4), a low molecular weight pterdine, is an essential cofactor required for the activity of NO synthase (NOS). Existing evidences show that lipopolysaccharide and proinflammatory cytokines can cause significant rises in bipoterin levels, which in turn augments the synthesis of NO. Also inhibition of biosynthesis of BH4 can decrease NO formation, implying that BH4 may be involved in the pathophysiological alterations of sepsis. However, the precise mechanisms of BH4 in regulating NO formation are not yet fully understood. In this review, we focus on the biological effects and regulation of BH4, as well as its potential role in sepsis. The therapeutic significance of biopterin synthesis inhibitors in septic symptoms is also discussed.

Influence of selective decontamination of the digestive tract on cell-mediated immune function and bacteria/endotoxin translocation in thermally injured rats.

OBJECTIVE: To determine the influence of pretreatment with selective decontamination of the digestive tract (SDD) on systemic immunosuppression, and the relationship between bacteria/endotoxin translocation and abnormalities of immune function in thermally injured rats. DESIGN, MATERIALS, AND METHODS: Animals were subjected to a 40% full-thickness scald injury, and divided into SDD-treated and control groups. The treatment group received SDD (polymyxin E, tobramycin, and 5-flucytosine) by gavage twice daily for 3 days before the experiment and continued for 5 days after thermal injury. The control group was given the same amount of water. The parameters reflecting cell-mediated immunity, including splenocyte proliferation in response to mitogens, interleukin 2 (IL-2) production, and lymphocyte subpopulation, were measured before injury and 1 and 5 days after burn, respectively. MEASUREMENTS AND MAIN RESULTS: Thermal injury resulted in marked reduction in splenocyte proliferative response to T-cell mitogens, IL-2 production, and T-helper/suppressor cells (CD4/CD8) ratio. Prophylactic treatment with SDD significantly decreased the incidences of bacterial translocation and endotoxemia, prevented suppressive mitogenic response and inadequate IL-2 production (p < 0.05-0.01) but did not affect the abnormal ratio of CD4 to CD8 T lymphocytes in blood (p > 0.05). CONCLUSIONS: These results suggest that bacteria/endotoxin translocation from the gut appears to be involved in cell-mediated immune dysfunction as a consequence of thermal injury. Pretreatment with SDD might attenuate postburn immunosuppression by preventing translocation events.

Reduction of circulating prostaglandin E2 level by antiserum against core lipopolysaccharide in a rabbit model of multiple organ failure.

OBJECTIVE: To determine the potential role of prostaglandin E2 (PGE2) in the development of multiple organ dysfunction or failure (MOF), the possible effects of antiserum directed against Re chemotype lipopolysaccharide (LPS, from Re mutant of Escherichia coli F515) on circulating PGE2 level and survival rate, and whether there is an elevation in the plasma LPS concentration that could account for the induction of arachidonic acid metabolite in a rabbit model of MOF caused by acute hypovolemic insult. DESIGN, MATERIALS, AND METHODS: An animal model of MOF in rabbits, engendered by feeding live Escherichia coli O111:B4 before hemorrhagic shock (35-40 mm Hg for 60 min), was used in the present study. Re-LPS antiserum was given intravenously in the treatment group at the onset of hemorrhage and 4 hours after resuscitation. The animals that received equal volumes of normal rabbit serum and antiserum served as the control group. MEASUREMENTS AND MAIN RESULTS: The circulating PGE2 level was not increased at the end of shock (p > 0.05), but it was found to be significantly elevated 24 hours after hemorrhage and resuscitation in both groups. However, Re-LPS antiserum administration markedly decreased peak PGE2 level (p < 0.05) and attenuated multiple organ damage caused by acute insult. Concomitantly, there were also lower LPS concentrations in the treatment group as compared with the control group (p < 0.05-0.01). The survival rate was significantly increased in antiserum-treated rabbits 96 hours postinjury (treatment vs. control: 58.0% vs. 11.1%, p < 0.01). CONCLUSIONS: The results suggest that an excessive generation and release of PGE2 may be involved in the pathogenesis of immunosuppression and MOF following hemorrhage and resuscitation. Re-LPS antiserum has an inhibitory effect on overproduction of circulating PGE2 and the ability to improve survival with MOF. Gut-derived endotoxemia, bacterial translocation, or both, could account, at least in part, for the PGE2 formation and release in animals response to acute hypovolemic insult.

[An experimental study of the mechanism of shock-induced enterogenic infection after long-term standard total parenteral nutrition].

In this study, gnotobiotic rats were subjected to total parenteral nutrition (TPN) and subsequent hemorrhagic shock to study the mechanism of enterogenic infection in these circumstances. (1) Long-term (8-12 days) TPN induced impairment of gut barrier function, evidenced by atrophy of intestinal mucosa, significant decrease in diamine oxidase activity of intestinal mucosa and blood, and marked micro-ecologic imbalance of the intestinal mucosa flora with dominant growth of aerobes and relative decrease in anaerobes. The degree of mucosal damage were proportional to the duration of TPN. (2) In TPN+shock groups, further damage was found in the mucosa, with a large number of invading gram-negative organisms and a significant decrease in DAO activity as compared to that with TPN only groups. These changes were significantly correlated with enhanced bacterial translocation, elevation of LPS and MDA levels in the plasma. These findings suggested that long term TPN impairing gut barrier function, precipitated posttraumatic gut barrier failure. The determination of plasma DAO activity may help in the early diagnosis of gut injury during TPN and after trauma.

Protective effect of Re-LPS antiserum on experimental multiple system organ failure.

This study examines the possible beneficial effect of Re-LPS (F515) antiserum on experimental multiple system organ failure (MSOF) in rabbits. The results showed that the plasma LPS level was significantly decreased, and it took a shorter period to clear up LPS in experimental than in control rabbits after receiving Re-LPS antiserum. Pretreatment with antiserum can markedly improve the function of the liver, lungs, kidneys, blood and gastrointestinal tract. The MSOF incidence in the group of rabbits receiving immune sera was only 11.2% and the survival rate was raised by about 40.0%. The results suggest that early passive immunotherapy may neutralize gut-derived endotoxin, inhibit endotoxin-induced mediators release and prevent development of severe complications due to sepsis. It is therefore postulated that LPS core antiserum may provide a prophylactic effect on the development of experimental MSOF.

[Bacterial translocation from the gastrointestinal tract and endogenous infection induced by immunosuppression after burn].

This study described the endogenous infections and bacterial translocation from GI tract caused by immunosuppression after burn. In the group of burned plus injected dexamethasone (DXM) (BIS, n = 31), the rate of enteric bacteria translocation was 67.4%, the rate of visceral abscess was 65.5%, much more higher than in the group of only DXM (IS, n = 15). The translocation of intestinal bacteria also was found in the group of only burned (BU 3/15) and control (ck, 1/23), but endogenous infection did not occur in both group. The bacteria cultured from the rat organs are mainly enteric bacilli and corynebacterium. It was assumed that the endogenous infection was originated from the conditions in which the micro-ecologic system of indigenous intestinal flora was disturbed, immunologic function was suppressed by the overlapped effect of burn and injection of DXM, the biological antagonism among the intestinal flora was attenuated, and the intestinal bacilli overgrew, passed through the epithelia of intestine into lymphatic vessel and mesenteric lymph nodes, then colonized and multiplied in other organs, resulting in endogenous infection.

Active oxygen in neuromuscular disorders.

Although muscle and nerve are reasonably well protected against active oxygen and related free radicals, environmental or inherited malfunctions can overpower their defences. Active oxygen is involved in many neuropathies and myopathies. In every case the damage is caused by agents which exert effects disproportionately greater than the quantities encountered, through a variety of amplification mechanisms. We can categorize these amplification mechanisms as follows: (a) non-replacement of targets (e.g. loss of genetic information, ataxia telangectasia being an hereditary ataxia in which an oxygen mediated chromosomal instability is apparent), (b) non-removal of unwanted materials (e.g. lipofuscin accumulation in brain and heart), (c) redox cycling, usually involving catalysis by trace-metal ions (e.g. some forms of Parkinsonism), (d) non-redox catalysis (e.g. toxicity in cardiac muscle or brain due to vanadium or aluminium respectively), (e) modification of ion transport (e.g. calcium ionophore or acrylamide induce histopathological changes in muscle, similar in some respects to those seen in Duchenne muscular dystrophy), (f) compromised defences (e.g. muscle and nerve become particularly susceptible to free radical damage after loss of the protective actions of vitamin E), and (g) amplification by inflammatory and immune responses (e.g. multiple sclerosis, reperfusion injury to brain and heart, and traumatic injury to nervous tissue). Unfortunately, a variety of therapeutic agents which might be expected to protect against almost every conceivable form of oxygen mediated damage have proved clinically ineffective in most of these disorders. The reasons for this will be explored with an emphasis on common features, differences, mechanisms, and potential therapeutic approaches.