[Application value of neutrophil lymphocyte ratio combined with age in the diagnosis of dual-phenotype hepatocellular carcinoma].

Objective: This study retrospectively analyzed the clinical medical records of patients with dual phenotypic liver cancer (DPHCC) and those (non-DPHCC) in the same period to seek quick and effective biomarkers for differential diagnosis. Methods: A retrospective study was conducted on 164 patients who underwent radical hepatocellular carcinoma resection at Affiliated Hospital of Nantong University from May 2017 to May 2020, including 29 patients with DPHCC, accounting for 17.7% (age: 53.9±10.0). There were 135 non-DPHCC patients, accounting for 82.3% (age, 62.6±9.1). The clinical records of the above patients were collected,including the basic information of the patients, clinical symptoms and signs, history of infection, laboratory test indexes one day before surgery, postoperative pathological report and other relevant data, The follow-up time was 18 months and the data were complete. By analyzing the clinical data of DPHCC patients and non-DPHCC patients in the same period, to find quick and effective differential diagnostic indicators, and to explore the indicators indicating poor prognosis of DPHCC patients. Results: One-way ANOVA showed significant differences in age, AFP[143(4.8-984.8) vs 9.9(2.8-71.3) µg/L], NLR (3.650±1.924 vs 2.220±1.486), neutrophil count, lymphocyte count, vascular infiltration rate, TNM stage, Chinese Hepatocellular carcinoma Staging (CNLC), Child grade, and Japanese General Staging Score (JIS) (P<0.05). Multivariate logistic regression analysis identified age(OR score:0.967,95%CI:0.860-0.957) and NLR(OR score:1.564,95%CI:1.205-2.029) as independent risk factors for DPHCC differential diagnosis. The receiver operating characteristic(ROC) curve was used to evaluate the diagnostic efficiency of NLR, and the best cut-off value was 2.586. The combination of age at onset improved the efficiency of differential diagnosis. When reaching the maximum diagnostic efficiency, the area under curve(AUC) was 0.836, the sensitivity was 89.66%, and the specificity was 65.93%. Conclusion: NLR combined with the age of disease has certain feasibility in predicting DPHCC and may be an effective index to distinguish DPHCC from non-DPHCC patients.

Effects of non-drug interventions on depression, anxiety and sleep in COVID-19 patients: a systematic review and meta-analysis.

OBJECTIVE: Patients with Coronavirus Disease 2019 (COVID-19) suffer from anxiety, depression and sleep disorders due to isolation treatment, among other reasons. Whether non-drug interventions can be alternative therapies for COVID-19 patients with anxiety, depression and sleep disorders is controversial. Therefore, we conducted a meta-analysis and systematic review to evaluate the effects of non-drug interventions on anxiety, depression and sleep in patients with COVID-19 to provide guidance for clinical application. MATERIALS AND METHODS: We searched the following databases for randomized controlled trials (RCTs) from December 2019 to July 2020: China Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI), Chongqing VIP Chinese Science and Technology Periodical Database (VIP), Wanfang, Cochrane Library, Web of Science, PubMed, MEDLINE and Embase. Two investigators independently screened the literature according to the inclusion and exclusion criteria, extracted data and evaluated the risk of bias in the included studies. Meta-analysis was performed using RevMan5.3 software. RESULTS: A total of 5 articles with 768 subjects were included. Meta-analysis results indicated that non-drug interventions can reduce anxiety [SMD=-1.40, 95% CI (-1.62, -1.17), p<0.00001] and depression [SMD=-1.22, 95% CI (-2.01, -0.43), p=0.002] scores in patients with COVID-19. Descriptive analysis indicated that non-drug interventions can improve the sleep status of COVID-19 patients. Sensitivity analysis indicated that the meta-analysis results were stable. Egger's test and Begg's test showed no publication bias. CONCLUSIONS: This meta-analysis found that non-drug interventions can reduce the anxiety and depression scores of patients with COVID-19. Due to the limitations of this study, more high-quality studies are needed to verify the findings, especially the effect of non-drug interventions on improving the sleep status of COVID-19 patients.

[Short-term efficacy evaluation of laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy].

Objective: To investigate short-term efficacy of laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy. Methods: A case series study was carried out. Case inclusion criteria: (1) patient was confirmed as gastric adenocarcinoma by gastroscopic biopsy before operation; (2) locally advanced gastric cancer was confirmed by abdominal CT before operation; (3) no distant metastases such as liver, lung, and posterior peritoneal lymph nodes, and no tumor directly invading the pancreas, spleen, liver, and colon were verified by superficial lymph node ultrasound, chest and abdominal CT before operation;(4) total gastrectomy or proximal gastrectomy plus D2 lymphadenectomy were performed, and R0 resection was confirmed by postoperative pathology. Exclusion criteria: (1) intraperitoneal dissemination or distant metastasis was found during laparoscopic exploration; (2) No.10 lymph nodes were significantly enlarged or fused into clusters; (3) pathological diagnostic data were incomplete. According to above criteria, the clinicopathological data of 36 patients who underwent laparoscopic spleen-preserving No.10 lymphadenectomy and left epigastrium mesogastric excision based on interspace anatomy for advanced proximal gastric cancer in The First Affiliated Hospital of Zhengzhou University from June 2017 to March 2018 were retrospectively collected and analyzed. The intraoperative conditions, postoperative recovery and complications of patients were analyzed. Results: In 36 patients, the mean age was (59.8±8.0) years, the mean BMI was (23.9±3.5) kg/m(2), and 8 cases (22.2%) received preoperative chemotherapy. All the patients underwent successfully the laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision. In the examination of postoperative resected specimens, it was found that the mesangial boundary of the upper and posterior part of the stomach was smooth, indicating the efficiency of complete mesangial resection. No case was converted to open operation. The mean time of lymph node dissection and mesangial resection was (34.2±11.4) minutes. The mean blood loss during operation was (44.8±21.3) ml. The mean number of lymph node dissection per patient was 45.6±17.6. The mean number of No. 11p+11d lymph node dissection was 3.1± 2.8 per patient, and 7 patients were pathologically positive with metastasis rate of 19.4% (7/36). The mean number of No.10 lymph node dissection was 2.9±2.5 per patient, and 2 patients were pathologically positive with metastasis rate of 5.6% (2/36). The time to postoperative flatus was (3.8±0.6) days, time to removal of nasogastric was (1.9±0.7) days, time to the first intake of fluid was (3.0±0.4) days, time to removal of drainage tube was (6.0±1.2) days. Postoperative mean hospital stay was (12.8±4.0) days. One case (2.7%) developed pulmonary embolism and 1 case (2.7%) developed gastroplegia after operation. The morbidity of postoperative complication was 5.6% (2/36). No operative site infection, postoperative bleeding and death within postoperative 30-day were observed. All the 36 patients were followed up and the median follow-up was 18 months (12-28 months). Seven patients died of tumor relapse and metastasis (3 cases died within postoperative 1 year) and another 1 case developed colonic cancer 17 months after operation. Conclusion: Laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy is safe and feasible.

[Experts consensus on the management of delirium in critically ill patients].

To establish the experts consensus on the management of delirium in critically ill patients. A special committee was set up by 15 experts from the Chinese Critical Hypothermia-Sedation Therapy Study Group. Each statement was assessed based on the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) principle. Then the Delphi method was adopted by 36 experts to reassess all the statements. (1) Delirium is not only a mental change, but also a clinical syndrome with multiple pathophysiological changes. (2) Delirium is a form of disturbance of consciousness and a manifestation of abnormal brain function. (3) Pain is a common cause of delirium in critically ill patients. Analgesia can reduce the occurrence and development of delirium. (4) Anxiety or depression are important factors for delirium in critically ill patients. (5) The correlation between sedative and analgesic drugs and delirium is uncertain. (6) Pay attention to the relationship between delirium and withdrawal reactions. (7) Pay attention to the relationship between delirium and drug dependence/withdrawal reactions. (8) Sleep disruption can induce delirium. (9) We should be vigilant against potential risk factors for persistent or recurrent delirium. (10) Critically illness related delirium can affect the diagnosis and treatment of primary diseases, and can also be alleviated with the improvement of primary diseases. (11) Acute change of consciousness and attention deficit are necessary for delirium diagnosis. (12) The combined assessment of confusion assessment method for the intensive care unit and intensive care delirium screening checklist can improve the sensitivity of delirium, especially subclinical delirium. (13) Early identification and intervention of subclinical delirium can reduce its risk of clinical delirium. (14) Daily assessment is helpful for early detection of delirium. (15) Hopoactive delirium and mixed delirium are common and should be emphasized. (16) Delirium may be accompanied by changes in electroencephalogram. Bedside electroencephalogram monitoring should be used in the ICU if conditions warrant. (17) Pay attention to differential diagnosis of delirium and dementia/depression. (18) Pay attention to the role of rapid delirium screening method in delirium management. (19) Assessment of the severity of delirium is an essential part of the diagnosis of delirium. (20) The key to the management of delirium is etiological treatment. (21) Improving environmental factors and making patient comfort can help reduce delirium. (22) Early exercise can reduce the incidence of delirium and shorten the duration of delirium. (23) Communication with patients should be emphasized and strengthened. Family members participation can help reduce the incidence of delirium and promote the recovery of delirium. (24) Pay attention to the role of sleep management in the prevention and treatment of delirium. (25) Dexmedetomidine can shorten the duration of hyperactive delirium or prevent delirium. (26) When using antipsychotics to treat delirium, we should be alert to its effect on the heart rhythm. (27) Delirium management should pay attention to brain functional exercise. (28) Compared with non-critically illness related delirium, the relief of critically illness related delirium will not accomplished at one stroke. (29) Multiple management strategies such as ABCDEF, eCASH and ESCAPE are helpful to prevent and treat delirium and improve the prognosis of critically ill patients. (30) Shortening the duration of delirium can reduce the occurrence of long-term cognitive impairment. (31) Multidisciplinary cooperation and continuous quality improvement can improve delirium management. Consensus can promote delirium management in critically ill patients, optimize analgesia and sedation therapy, and even affect prognosis.

SPAG5 promotes proliferation and suppresses apoptosis in bladder urothelial carcinoma by upregulating Wnt3 via activating the AKT/mTOR pathway and predicts poorer survival.

Sperm-associated antigen 5 (SPAG5) is involved in various biological processes. However, the roles of SPAG5 in bladder urothelial carcinoma (BUC) are unknown. This study showed that upregulation of SPAG5 was detected frequently in primary BUC tissues, and was associated with significantly worse survival among the 112 patients that underwent radical cystectomy (RC). Up and downregulating the expression of SPAG5 enhanced or inhibited, respectively, the proliferation of BUC cells in vitro and in vivo, and suppressed or enhanced, respectively, apoptosis in vitro and in vivo. Moreover, SPAG5 increased the resistance of BUC cells to chemotherapy-induced apoptosis. Mechanistic investigations showed that SPAG5 promotes proliferation and suppresses apoptosis in BUC at least partially via upregulating Wnt3 through activating the AKT/mTOR signaling pathway. The importance of the SPAG5/AKT-mTOR/Wnt3 axis identified in BUC cell models was confirmed via immunohistochemical analysis of a cohort of human BUC specimens that underwent RC. Collectively, our data suggested that in patients with BUC who underwent RC, high SPAG5 expression is associated with poor survival. In addition, targeting SPAG5 might represent a novel therapeutic strategy to improve the survival of patients with BUC.

Rapamycin induces human acute promyelocytic leukemia cell HL-60 autophagic apoptosis.

OBJECTIVE: We aimed at investigating the effects of rapamycin on apoptosis and autophagy of human acute promyelocytic leukemia cell line HL-60, and to preliminarily explore the mechanism of extra medullary infiltration of leukemia cells with human acute promyelocytic leukemia cell line HL-60 as the object of study, providing a theoretical basis for the clinical treatment of leukemia. MATERIALS AND METHODS: After HL-60 cells were cultured in vitro, the effect of rapamycin on proliferation ability of HL-60 cells was determined by methyl thiazolyl tetrazolium (MTT) method, the cell apoptosis ratio was detected by flow cytometer, the change of autophagy after HL-60 cells acted by rapamycin was tested by monodansylcadaverine (MDC) fluorescence staining, the mRNA expression of autophagy-related molecule was detected by polymerase chain reaction (PCR), and the expressions of apoptosis-related protein and autophagy-related protein were determined by Western blotting (WB). RESULTS: HL-60 cell proliferation could be significantly inhibited by rapamycin (80 µg/mL-640 µg/mL), which was in a dose-dependent manner. HL-60 cell apoptosis ratio and apoptosis-related protein expression were distinctly improved by rapamycin. Cell autophagy level, mRNA expression of autophagy-related molecule and autophagy-related protein expression were remarkably induced by rapamycin. CONCLUSIONS: Rapamycin can induce HL-60 cell apoptosis, which is produced mainly by inducing cell autophagy.

Analysis on kidney injury-related clinical risk factors and evaluation on the therapeutic effects of hemoperfusion in children with Henoch-Schonlein purpura.

OBJECTIVE: To investigate risk factors related to kidney injury in children with Henoch-Schonlein purpura (HSP) and to study the therapeutic effects of hemoperfusion (HP) on kidney injury in HSP children, providing clinical evidence for early prevention and treatment of HSP. PATIENTS AND METHODS: Children who suffered from HSP for the first time were selected as study objects and they were followed up for 12 months. Single factor analysis and multi-factor Logistic regression analysis were performed for children's demographic characteristics (age, gender), clinical manifestations (rash duration time, rash recurrence times, digestive tract hemorrhage, abdominal pain, arthralgia, HSP recurrence) and laboratory indexes (peripheral blood WBC, PLT, ESR, CRP, serum IgG, serum IgA, IgM, serum C3, serum C4, TC, TG, HDL, LDL). Meanwhile, participants were divided into treatment group (HP treatment) and control group, and the protective effects of HP on renal function of HSP children were discussed. RESULTS: Single factor analysis indicated age ≥ 6 years, rash recurrence ≥ 3 times, rash duration time ≥ 1 month, digestive tract hemorrhage, peripheral blood PLT, WBC, serum TC and serum LDL levels had statistically significant differences between the two groups. Multi-factor Logistic regression analysis indicated rash recurrence ≥ 3 times, digestive tract hemorrhage, decline of peripheral blood PLT count, and increases of serum TC and LDL were closely related to kidney injury of HSP children. After discharge, kidney injury comparison between treatment group and control group in follow-up had a statistical difference. CONCLUSIONS: Rash recurrence ≥ 3 times, digestive tract hemorrhage, decline in peripheral blood PLT count, increases of serum TC and LDL, are risk factors of kidney injury in HSP children. HP can protect renal function of HSP children.

Asthma symptoms among Chinese children: the role of ventilation and PM10 exposure at school and home.

SETTING: Respiratory symptoms in relation to environment in Taiyuan, China. OBJECTIVE: To study associations between school/home exposure and prevalence of respiratory symptoms and self-reported respiratory infections (RIs) among students. DESIGN: A total of 2134 pupils from 10 schools answered a questionnaire; air pollution at school was measured. RESULTS: The prevalence of current wheeze, daytime attacks of breathlessness, nocturnal cough, nocturnal respiratory symptoms and RI was respectively 4.4%, 18.7%, 11.6%, 3.6% and 32.3%. Environmental tobacco smoke (ETS) at home was associated with daytime attacks of breathlessness (OR 1.41), nocturnal cough (OR 1.41) and RI (OR 1.26). Redecoration or indoor painting at home was associated with wheeze (OR 2.02), daytime attacks of breathlessness (OR 1.57) and nocturnal symptoms (OR 1.83). Dampness or mould at home was associated with daytime attacks of breathlessness (OR 1.68), nocturnal cough (OR 1.68) and RI (OR 1.69). Particulate matter of diameter  10 microns (PM10) in the classroom was associated with nocturnal cough (OR 1.20/10 µg/m3). PM10 outside school was associated with daytime attacks of breathlessness (OR 1.07/10 µg/m3) and nocturnal cough (OR 1.13/10 µg/m3). Indoor carbon dioxide, a marker of poor ventilation (OR 1.52/1000 parts per million), and relative humidity (RH) (OR 1.33/10%) were associated with nocturnal cough. CONCLUSION: Dampness, redecoration and ETS at home, as well as poor ventilation flow, and high RH and PM10 concentration at school may influence respiratory health.

[Three-dimensional circulation perfusion culture of hepatocytes in the liver decellularizedscaffold].

Objective: The intact rat liver decellularized scaffolds were preparedand repopulated hepatocytes by continuous perfusion technology.Toprovideexperimental support for the application of decellularized liver scaffolds in liver engineering. Method: Decellularized liver scaffolds were obtained by perfusing method. The composition and structure was examined by HE, Masson, Sirius red stain and immunofluorescence. The ultrastructure was examined by scanning electron microscope (SEM). DNA content was used to confirm the effect of decellularization. The circulation perfusion device was established. Hepatocytes were recellularized into the scaffolds by multiposition parenchymal injection method and infusion method, thenthe scaffolds were cultured in the circulation perfusion device in vitro. After cultivation, HE staining, immunofluorescence and SEM were conducted to observe the growth situation of hepatocytes in the scaffolds. Results: The rat decellularized liver scaffolds were successfully obtained by perfusion method. Histological staining demonstrated the remove of cellular component and the reservation of extracellular cellmatrix. Immunofluorescence staining demonstrated the retention of collagen I. SEM showed that the ultrastructure of the extracellular cell matrix presented thereticular structure. DNA content of the scaffolds was 47.5±18.1 ng/mg. The circulation perfusion device was composed of a peristaltic pump, oxygenator, chamber and the convey tubes. The multipositional parenchymal injection method resulted in a better engraftment rate. HE staining, immunofluorescence and SEM revealed that the growth and function of hepatocytes were goodin the scaffold. Conclusion: The decellularized rat liver scaffolds have favorable biochemical properties. The liver decellularized scaffolds applied with the circulation perfusion device could provide a well 3D plat form for culture of hepatocytes.

[PM2.5 and polluted gas exposure induced acute injury in rats'larynx membrane].

Objective:To study acute injury of rat's larynx membrane which were exposured to PM2.5 and polluted gas in serious air pollution. Method:Rats were divided into 3 groups of 5 animals each, the normal control group, gas exposured group, PM2.5 +gas exposured group. The control group served as the normal healthy control, rats of the gas exposured group had been exposured polluted gas in the A cabin, rats of PM2.5 +gas exposured group had been exposured PM2.5 and polluted gas in the B cabin. The total exposure time was six days. We collected specimen from 3 groups immediately after exposure. Histopathological changes in the rat's larynx membrane were assessed using hematoxylin and eosin staining(HE), the expression of IL-1ß in laryngeal mucous membrane were assessed using immunohistochemical staining(IHC), and the ultrastructure changes were observed under TEM. Result:HE staining showed the rat's laryngeal mucosa cells in the normal control group had no obvious abnormalities, laryngeal mucosa of rats in the two experimental group had different degrees of cell proliferation, cell polarity change, etc. IHC staining showed that there's no obvious expression of IL-1ß in laryngeal mucous membrane in the normal control group, and positive expression in the two experimental group. TEM showed the control group had no obvious abnormalities, the laryngeal mucosa cells in the two groups of experimental groups had been found irregular nucleus and damaged mitochondria. Conclusion:Exposed to PM2.5 and polluted gas in severe atmospheric pollution 6 days, the acute damage in rats laryngeal mucosa can be observed.

Role of inflammatory responses in the pathogenesis of human cerebral aneurysm.

We aimed to detect expressional profiles of intracellular cell adhesion molecule-1 (ICAM-1), nuclear factor-kappa B (NF-κB), and monocyte chemoattractant protein-1 (MCP-1) in human cerebral aneurysm, in order to investigate the effect of chronic inflammation on the pathogenesis of cerebral aneurysm. Samples from 40 cases of human cerebral aneurysms diagnosed at our hospital were selected along with 20 normal cerebral artery samples. Western blotting and immunohistochemical (IHC) staining were used to reveal expressional profiles of ICAM-1 and NF-κB in the aneurysmal wall of patients and normal cerebral artery tissues. Reverse transcription (RT)-PCR was employed to detect changes in transcript levels of MCP-1 mRNA. Western blotting showed significantly higher expressions of ICAM-1 and NF-κB in patients with cerebral aneurysm compared to the normal group (P < 0.01), which was consistent with IHC staining results. RT-PCR revealed significantly higher MCP-1 transcripts in cerebral aneurysm tissues compared to the normal group (P < 0.01), in addition to a positive relationship between ICAM-1 and NF-κB expression levels. In conclusion, expression levels of ICAM-1, NF-κB, and MCP-1 in patients are significantly elevated, suggesting an enhanced chronic inflammatory response and a significant correlation between inflammatory factors/adhesion molecules and the pathogenesis of cerebral aneurysm.

Epithelial-mesenchymal transition biomarkers and support vector machine guided model in preoperatively predicting regional lymph node metastasis for rectal cancer.

BACKGROUND: Current imaging modalities are inadequate in preoperatively predicting regional lymph node metastasis (RLNM) status in rectal cancer (RC). Here, we designed support vector machine (SVM) model to address this issue by integrating epithelial-mesenchymal-transition (EMT)-related biomarkers along with clinicopathological variables. METHODS: Using tissue microarrays and immunohistochemistry, the EMT-related biomarkers expression was measured in 193 RC patients. Of which, 74 patients were assigned to the training set to select the robust variables for designing SVM model. The SVM model predictive value was validated in the testing set (119 patients). RESULTS: In training set, eight variables, including six EMT-related biomarkers and two clinicopathological variables, were selected to devise SVM model. In testing set, we identified 63 patients with high risk to RLNM and 56 patients with low risk. The sensitivity, specificity and overall accuracy of SVM in predicting RLNM were 68.3%, 81.1% and 72.3%, respectively. Importantly, multivariate logistic regression analysis showed that SVM model was indeed an independent predictor of RLNM status (odds ratio, 11.536; 95% confidence interval, 4.113-32.361; P<0.0001). CONCLUSION: Our SVM-based model displayed moderately strong predictive power in defining the RLNM status in RC patients, providing an important approach to select RLNM high-risk subgroup for neoadjuvant chemoradiotherapy.

A serum proteomics approach to the diagnosis of ectopic pregnancy.

An ectopic pregnancy (EP) occurs when implantation of the embryo occurs outside of the uterus. If left untreated, the developing fetus will continue to grow, leading to life-threatening consequences for the mother. A major difficulty with the diagnosis of ectopic pregnancy is that methods of detection are limited, and some, such as ultrasound, are not very reliable in the earliest days of gestation. Currently, no effective serum test exists to distinguish an ectopic pregnancy from a normal intrauterine pregnancy. The incidence of ectopic pregnancy is increasing and has doubled in the last 20 years. It is now the second most common cause of maternal death in the first trimester of pregnancy. To address this issue, we initiated a project to identify serum markers of ectopic pregnancy. The subjects for these studies presented at the Hospital of the University of Pennsylvania. We obtained over 140 serum samples from women with suspected ectopic pregnancy: women presenting with pain and/or bleeding in the first trimester of pregnancy. The approximate racial breakdown of the subjects is as follows: African American, 36%; Caucasian, 3%; Asian, 2%; Hispanic, 1%; unknown, 58%. Serum samples from 139 women (62 with ectopic pregnancy and 77 with a normal intrauterine pregnancy) were applied to WCX2 (weak ion exchange) protein chip surfaces and analyzed for serum markers using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Several proteins in the 7500-18,000 Da mass range were identified that may discriminate an ectopic pregnancy from an intrauterine pregnancy. The most promising markers were analyzed using classification and regression tree analysis (CART) with and without clinical variables (serum hCG value, length of amenorrhea). Two different algorithms were developed that classify the patients on the basis of sensitivity (number of EPs who screen positive/# of EPs) or specificity (# of healthy patients who screen negative/# of healthy). Our current approach is to refine these two "rule sets" to segregate patients into three groups: those who need immediate intervention for a probable ectopic pregnancy, those who appear to have a normal pregnancy, and those who need further monitoring for diagnosis.

A new method for characterizing denitrifying phosphorus removal bacteria by using three different types of electron acceptors.

This study investigated the characteristics of denitrifying phosphorus removal bacteria by using three different types of electron acceptors as well as the positive role of nitrite in phosphorus removal process. Denitrifying phosphorous removal bacteria was enriched under anaerobic-anoxic (A/A) condition. To understand A/A sludge better, sludge from two other sources were also studied. These include sludges obtained from a lab-scale anaerobic-anoxic-aerobic (A/A/O) system and a local sewage treatment plant. Three types of possible electron acceptors (oxygen, nitrate and nitrite) were examined for their roles in phosphorus uptake. The results obtained indicated that oxygen, nitrate and nitrite were able to act as electron acceptors successfully. This observation suggested that in addition to the two well-accepted groups of phosphorus removal bacteria (one can only utilize oxygen to take up phosphorus, P(O), while the other can use both oxygen and nitrate, P(ON)), a new group of phosphorus removal bacteria, P(ON(n)), which could use oxygen, nitrate or nitrite to take up phosphorus was identified. The relative population of these three types of bacteria could be calculated from results obtainable from phosphorus uptake batch experiments with either oxygen or nitrate or nitrite as electron acceptor. The results obtained in this study showed that A/A sludge had similar phosphorus removal performance as the A/A/O sludge. However, it has better denitrifying phosphorus removal capability, which was demonstrated by the relative population of the three groups of bacteria. The results also suggested that nitrite was not an inhibitor to phosphorus removal process. Instead, it is an alternative electron acceptor to oxygen or nitrate.

Proteomic analysis of nipple aspirate fluid to detect biologic markers of breast cancer.

The early detection of breast cancer is the best means to minimise disease-related mortality. Current screening techniques have limited sensitivity and specificity. Breast nipple aspirate fluid can be obtained noninvasively and contains proteins secreted from ductal and lobular epithelia. Nipple aspirate fluid proteins are breast specific and generally more concentrated than corresponding blood levels. Proteomic analysis of 1 microl of diluted nipple aspirate fluid over a 5-40 kDa range from 20 subjects with breast cancer and 13 with nondiseased breasts identified five differentially expressed proteins. The most sensitive and specific proteins were 6500 and 15 940 Da, found in 75-84% of samples from women with cancer but in only 0-9% of samples from normal women. These findings suggest that (1) differential expression of nipple aspirate fluid proteins exists between women with normal and diseased breasts, and (2) analysis of these proteins may predict the presence of breast cancer.

[The analysis of rice total RNA by capillary electrophoresis].

When the RNase was eliminated through backing at 200 degrees C or treated by diethylpyrocarbonate (DEPC), the total RNA of rice could be separated within 15 min using 1.0%T, 0%C linear polyacrylamide as sieving matrix and 7 mol/L urea as denaturant. The tRNA of rice can be separated into two classes and about nine peaks when high concentrated polyacrylamide sieving matrix (5.0%T, 0%C) was used. This technique could provide one of the rapid and accurate methods for the determination of RNA in plants.

[Determination of human PDGF-B promoter binding nuclear protein by capillary electrophoresis].

With the use of 1.0% T, 0% C linear polyacrylamide as sieving matrix, 0.25 x TBE(Tris 89 mmol/L, boric acid 89 mmol/L, EDTA 2 mmol/L) as running buffer and 15 degrees C as column temperature, the human PDGF-B promoter binding nuclear protein can be determined within 50 min with good resolution. The results proved that there are two proteins having strong ability binding human PDGF-B promoter, which similar to that in slab gel electrophoresis. This technique can provide one of the rapid and accurate separation methods in the studying of the formation and repression behavior of DNA binding protein based on PDGF gene as the target.

Molecular determinants of the caspase-promoting activity of Smac/DIABLO and its role in the death receptor pathway.

Smac/DIABLO is a mitochondrial protein that is released along with cytochrome c during apoptosis and promotes cytochrome c-dependent caspase activation by neutralizing inhibitor of apoptosis proteins (IAPs). We provide evidence that Smac/DIABLO functions at the levels of both the Apaf-1-caspase-9 apoptosome and effector caspases. The N terminus of Smac/DIABLO is absolutely required for its ability to interact with the baculovirus IAP repeat (BIR3) of XIAP and to promote cytochrome c-dependent caspase activation. However, it is less critical for its ability to interact with BIR1/BIR2 of XIAP and to promote the activity of the effector caspases. Consistent with the ability of Smac/DIABLO to function at the level of the effector caspases, expression of a cytosolic Smac/DIABLO in Type II cells allowed TRAIL to bypass Bcl-xL inhibition of death receptor-induced apoptosis. Combined, these data suggest that Smac/DIABLO plays a critical role in neutralizing IAP inhibition of the effector caspases in the death receptor pathway of Type II cells.

Helicobacter pylori increases proliferation of gastric epithelial cells.

The direct and indirect effects of helicobacter pylori on cell kinetics of gastric epithelial cell line AGS were investigated by flow cytometric analysis of Ki-67 positive cells and by MTT assay. Flow cytometric analysis of Ki-67 positivity permits detection of cells that are in S-phase, whereas the MTT assay is a colometric measure of the number of viable cells. In the absence of added stimulants, 23.06 (4.88)% mean (SD) of AGS cells were Ki-67 positive. When cells were preincubated in the presence of H pylori, there was a significant increase in Ki-67 positivity (66.20 (7.89)%, p < 0.001). This increase was not seen in cells cultured in the presence of Campylobacter jejuni (24.63 (8.11)% or Escherichia coli (21.66 (9.78)%). Pre-incubation of AGS cells with supernatants from both H pylori and mitogen activated peripheral blood lymphocytes also increased the per cent of cells that were Ki-67 positive (72.93 (8.68) and 69.96 (12.35)%; p, 0.001) respectively. Similar results were also found in MTT assay. These data show that both H pylori directly and the immune/inflammatory response to H pylori indirectly can influence the rate of epithelial cell proliferation, suggesting this bacterium may be an initiating step in gastric carcinogenesis and an important co-carcinogenic factor in H pylori positive subjects.