Nano-LC-MS/MS for the quantitation of prostanoids in immune cells.

Prostanoids, derivatives of arachidonic acid, are involved in inflammation and immune reactions. To understand the role of prostanoids produced by diverse immune cells, a highly sensitive quantitation method for prostaglandin E2 (PGE2), prostaglandin D2 (PGD2), 6-keto prostaglandin F1α (6-keto PGF1α), prostaglandin F2α (PGF2α), and thromboxane B2 (TXB2) by means of nano-liquid chromatography-tandem mass spectrometry has been developed. It was validated according to the guidelines of the Food and Drug Administration (FDA) in terms of linearity, precision, accuracy, recovery, stability, and lower limit of quantitation (LLOQ). The LLOQ were 25 pg/mL in the injected solution (75 fg on column (o.c.)) for PGE2 and PGD2 and 37.5 pg/mL (112.5 fg on column) for 6-keto PGF1α, PGF2α, and TXB2, respectively. It was successfully applied to murine mast cells isolated from paws after zymosan injection and to CD4(+) and CD8(+) T lymphocytes from blood of sensitized versus non-sensitized mice in context of a delayed type hypersensitivity model. About 5,000 (T cells) to 40,000 (mast cells) cells were sufficient for quantitation. In the mast cells, the production of PGE2 increased at a significantly higher extent than the synthesis of the other prostanoids. The T lymphocytes did not show any difference in prostanoid production, no matter whether they were obtained from sensitized mice or non-sensitized mice.

Effectiveness of fast-track rehabilitation vs conventional care in laparoscopic colorectal resection for elderly patients: a randomized trial.

AIM: The aim of the study was to evaluate the efficacy and safety of fast-track rehabilitation in elderly patients over 65 years of age, following laparoscopic surgery to remove colorectal cancer. METHOD: A total of 78 elderly patients with colorectal cancer who underwent laparoscopic colorectal resection were randomly assigned to receive either the fast-track care programme (n = 40) or the conventional perioperative care protocol (control group, n = 38). Medical personnel conducting the study were blinded to patients' clinical outcomes prior to statistical analysis. The fast-track protocol included no preoperative mechanical bowel irrigation, immediate oral alimentation and earlier postoperative ambulation exercise. The length of postoperative hospital stay, the length of time to regain bowel function and the rate of postoperative complications were compared between the two groups. RESULTS: The length of time to regain bowel function, including the passage of flatus[31 (26-40) h vs 38 (32-51) h, P = 0.001], to the first bowel movement [55 (48-63) h vs 64 (48-71) h, P = 0.009] and to start a liquid diet (12 [11-16] h vs 47 [35-50] h, P = 0.000) were significantly shorter in patients receiving the fast-track care protocol compared with those receiving the conventional care protocol. A shorter duration of postoperative hospital stay was recorded in patients receiving the fast-track program than in those receiving conventional care (P = 0.0001). A reduced percentage of patients who developed general complications was also observed in the fast-track group (5.0%vs 21.1%, P = 0.045). CONCLUSION: This randomized controlled trial has shown that in the elderly undergoing laparoscopic colorectal surgery, the fast-track recovery programme resulted in a more rapid postoperative recovery, earlier discharge from hospital and fewer general complications compared with a conventional postoperative protocol.

The contraceptive effect of a novel filtering-type nano-copper complex/polymer composites intra-vas device on male animals.

Contraceptive techniques which target vas deferens have been paid great attention for their good efficacy, safety and reversibility. We have made a filtering-type intra-vas device (IVD) using nano-copper complex/polymer composites. Twenty male adult Beagle dogs and 40 male rabbits were randomly assigned to four groups (sham-operation, IVD, reversal and vasectomy groups). Dogs' semen parameters, concentration of α-glucosidase, copper and zinc ions were tested pre-operation and 1, 3, 6 and 12 months post-operation. The pregnancy rates of the rabbits were evaluated by mating trials after the IVDs were implanted. The histology of testis, epididymides and vas deferens of the animals was examined using an electron microscope. Apoptosis of the cells in the testes, epididymides and vas deferens was detected by TUNEL method. There was no sperm in the semen of dogs, which had been inserted IVD and vasectomized at 1, 3, 6 and 12 months post-operation. The concentration of α-glucosidase in the IVD group, reversal group and sham-operation group was not significantly different between pre- and post-operation. The pregnancy rates of the female rabbits in the vasectomy, IVD and reversal groups were all zero, but the pregnancy rate in the reversal group, after taking out IVD, and that of the sham-operation group was 60% and 80%, respectively. The ultrastructures of the testes, epididymides and vas deferens of the male animals in the IVD group and sham-operation group were in normal ranges compared with the vasectomy group. The apoptosis of the cells in the testes, epididymides and vas deferens in the vasectomy group of both dogs and rabbits was obvious compared with the other groups. No significant changes in the quantities of copper and zinc ions were found in semen of the male dogs both pre- and post-operation. Our studies demonstrated that the filtering-type nano-copper complex/polymer composites intra-vas device may be an efficacious, safe and reversible male contraceptive device.

The clinicopathological and prognostic value of programmed death-ligand 1 in colorectal cancer: a meta-analysis.

INTRODUCTION: Programmed death-ligand 1 (PD-L1) is reportedly expressed in colorectal tumors. However, the prognostic role of PD-L1 in colorectal cancer (CRC) remains controversial. Therefore, we performed a meta-analysis to investigate the clinicopathological and prognostic impact of PD-L1 in CRC. METHODS: A comprehensive search in PubMed, Embase, the Cochrane Library, Web of Science and the ClinicalTrials.gov for publications about PD-L1 expression in colorectal cancer was done. The correlation between PD-L1 expression and clinicopathological features or survival outcomes was analyzed by odds ratios (OR) or hazard ratios (HR), at 95% confidence intervals (CI). RESULTS: The results show that the pooled HR of (1.34, 95% CI 1.02-1.65, p = 0.01) indicated the association of PD-L1 expression with overall survival (OS) in CRC patients. Meanwhile, the expression of PD-L1 was positively correlated with the lymph node metastasis (OR: 0.70, 95% CI 0.51-0.95, p = 0.00), gender (OR: 0.86, 95% CI 0.76-0.98, p = 0.05) and tumor location (OR: 1.39, 95% CI 1.14-1.71, p = 0.12). CONCLUSIONS: These results suggest that high expression of PD-L1 is associated with low OS in CRC. High PD-L1 expression may act as a negative factor for patients with CRC and help to identify patients suitable for anticancer therapy.

Factors associated with a clinician's decision to stop anti-tuberculosis treatment before completion.

OBJECTIVES: To investigate the diagnosis of pulmonary tuberculosis (PTB) and factors associated with a clinician's decision to stop anti-tuberculosis treatment before completion. DESIGN: The medical charts of all citizens of Taipei City, Taiwan, reported to have received treatment for PTB in 2003 were investigated. RESULTS: Of 1126 PTB patients, 512 (45.5%) started treatment immediately based solely on chest X-ray (CXR) findings; treatment for 214 (19.0%) was based on a positive sputum smear for acid-fast bacilli, for 261 (23.2%) it was based on other findings and for 139 (12.3%) it was based on a positive mycobacterial culture. Of the 1126 PTB patients, 156 (13.9%) had their diagnosis of TB changed by a clinician. Multivariate analysis shows that patients whose diagnosis was based on CXR or other findings, female patients, patients who interrupted treatment for 2 months, patients who continued care at other health facilities (transfer) and patients with lung cancer were significantly more likely to have their diagnosis changed than other groups. CONCLUSION: A substantial proportion of patients were prescribed anti-tuberculosis treatment based on CXR findings alone, and a considerable proportion were advised to stop treatment before completing a full course, findings that require the immediate attention of Taiwan's National Tuberculosis Programme.

Tuberculin reactivity in adults after 50 years of universal bacille Calmette-Guerin vaccination in Taiwan.

We aimed to assess whether tuberculin reactivity in adults is affected by bacille Calmette-Guerin (BCG) vaccination after 50 years of universal BCG vaccination with 80-95% coverage. A community-based study on tuberculin reactivity in 619 participants was conducted in February 2000 in Keelung city, Taiwan. Information on BCG vaccination policies and annual risk of infection (ARI) in the underlying population was extracted from consecutive national prevalence surveys relating to the period 1952-1997. Compared with the expected ARI estimate, the standardized morbidity ratio of positive tuberculin response for vaccination in infancy was 2.2 (95% CI 0.3-15.5) for those aged <10 years. The corresponding figures for older age groups ranged from 3.6 (95% CI 2.2-5.9) for those aged 10-12 years to 0.7 (95% CI 0.5-0.9) for those aged 57-67 years. This suggests that the effect of BCG vaccination on positive tuberculin response in adults aged >30 years is probably negligible irrespective of age at vaccination or revaccination and that the tuberculin skin test can be used to diagnose TB in control programmes in countries with moderate or high incidence of TB.

Opioid mediated effects on the immune system: sympathetic nervous system involvement.

Opioids have been hypothesized to suppress parameters of immune function by acting within the central nervous system to increase the activity of the hypothalamic-pituitary-adrenal axis and the sympathetic nervous system. Production of catecholamines and adrenocorticoids have been demonstrated to be responsible for many of the observed immunomodulatory effects which occur following opioid administration. In general, the sympathetic nervous system has been shown to play a role in regulating lymphocyte proliferation and natural killer cell activity as well as several other parameters of immune function. Here, we will focus primarily on the role of the sympathetic nervous system in modulating opioid induced immunosuppression. The role of the hypothalamic-pituitary adrenal axis is reviewed elsewhere in this issue.

The impact of national health insurance on the notification of tuberculosis in Taiwan.

SETTING: General notification of tuberculosis in Taiwan. OBJECTIVES: To ensure the completeness of notification of tuberculosis (TB) in Taiwan, the Bureau of National Health Insurance (NHI) introduced two policies in 1997: 1) the no-notification-no-reimbursement (NNNR) policy, and 2) the notification-fee (NF) policy. The goals of this study were to investigate the impact of the NNNR and NF policies on notification of TB. DESIGN: Review of all cases notified to the National TB Register from 1995 to 1999 to determine calendar trend, type of case and source of notification. RESULTS: There were 11,453 and 13,612 reported cases in 1995 and 1996, respectively. Following the implementation of the NHI policies, there was a 47% increase in 1997, with 20 021 reported cases. Quarterly reporting of cases reached a historic peak in the third quarter of 1997. The increase in reported cases was mainly from general hospitals/clinics. Since 1998, the number of reported cases has declined steadily, at a rate of 7% and 3% in 1998 and 1999, respectively. CONCLUSIONS: The NNNR and NF policies had a significant impact on notification of TB in Taiwan. These policies substantially improved completeness of reporting, an observation with implications for surveillance of other reported diseases.

Comparative evaluation of the BACTEC MGIT 960 system with solid medium for isolation of mycobacteria.

SETTING: The utilisation of new, rapid methods of diagnosis of tuberculosis is currently of great interest for tuberculosis control. This study was carried out in a teaching hospital in the eastern region of Taiwan. OBJECTIVE: The BACTEC MGIT 960 system was evaluated and compared with Lowenstein-Jensen (LJ) medium and Middlebrook 7H11 plate for recovery rate and time to detection of mycobacteria. DESIGN: A total of 1396 sputum samples were tested for the presence of mycobacteria. Specimens were processed and inoculated separately in the BACTEC MGIT 960 system, on LJ medium and 7H11 for comparative study. RESULTS: The BACTEC MGIT 960 detected 235 isolates (100%), followed by LJ with 205 isolates (87.2%) and 7H11 with 178 isolates (75.7%). The mean time to detection of Mycobacterium tuberculosis complex was 11.6 days with MGIT 960, 20.1 days with LJ, and 18.7 days with 7H11. The contamination rates were 15.1% with MGIT 960, 10.1% with LJ and 9.7% with 7H11. CONCLUSION: The BACTEC MGIT 960 system is a sensitive, rapid mycobacterial culturing system. However, the high contamination rate is a concern that should be carefully evaluated in the clinical setting.

Pulmonary resection in the treatment of patients with pulmonary multidrug-resistant tuberculosis in Taiwan.

SETTING: Chronic Disease Control Bureau, Department of Health, Taiwan. OBJECTIVE: To evaluate the role of pulmonary resection in the treatment of pulmonary tuberculosis resistant to isoniazid and rifampin (MDR-TB). DESIGN: In a retrospective cohort study, 27 MDR-TB patients who underwent pulmonary resection between December 1990 and March 1999 were reviewed. Individually-tailored treatment regimens were selected at a once-weekly staff conference following review of the patient's case history and drug susceptibility results. Surgery was performed for selected patients, essentially those: 1) whose medical treatment had failed, or for whom treatment failure seemed highly likely, or for whom post-treatment relapse seemed likely, 2) with predominantly localised disease, 3) with adequate cardiopulmonary reserve, and 4) whose treatment regimen had been composed of at least two effective drugs to diminish the mycobacterial burden. RESULTS: There was no surgical mortality apart from one peri-operative death (4%). Three patients (11%) developed complications, and 24 (92%) patients demonstrated sputum conversion and/or remained negative after surgery. Twenty-three patients have already completed treatment, and during a mean of 42 +/- 18 follow-up months (range 15-80 months), one patient relapsed. This patient was disease-free after another course of treatment. CONCLUSION: For selected patients, pulmonary resection may improve the outcome of pulmonary MDR-TB.

Centrally-mediated opioid-induced immunosuppression. Elucidation of sympathetic nervous system involvement.

Opioid-induced modulation of the immune system is a complex phenomenon involving opioid receptors, central and sympathetic neural pathways, catecholamine receptors, and other regulatory mechanisms. The precise neural pathways involved in centrally-mediated immune modulation are not currently defined. In addition, the physiological purpose for endogenous opioid modulation of the immune system is not well understood. Perhaps this modulation phenomenon represents an integral feedback loop within a much larger homeostatic control system. Indeed, the role of the HPA axis in immune regulation can not be discarded, and in fact, probably serves to balance immune function, in concert with multiple feedback systems, around some undiscovered parameter of efficiency. Perhaps the physiological role of endogenous opioid control is to act as a monitor poised to subvert chronic inflammatory processes and autoimmune disorders. Regardless of the evolutionary heritage of this and despite the overwhelming complexity of immune regulation, important work substantiating a bidirectional communication link between the brain and the immune system has created a foundation for further elucidation of the intricacies of immunoregulation.

Suppression of hepatic natural killer activity by liver metastasis of cancer and restoration of killer activity by oral administration of a Basidomycetes-derived polysaccharide, PSK.

PSK (Krestin) is a protein-bound polysaccharide with antitumor and immunomodulatory activity. In this study, the effects of the oral administration of PSK were investigated on the natural killer (NK) activity of liver-associated lymphocytes and their subfractions separated by density gradient centrifugation, in WKAH rats with liver metastasis of KDA hepatoma. PSK was administered orally, at a dose of 500 mg/kg once a day for 3 weeks. The NK activity of nonparenchymal liver cells (NPLC) and their subfractions, including large granular lymphocytes (LGL), was markedly augmented by this treatment. The effects of oral PSK were also examined in CDF1 mice with liver metastases of Colon 26 adenocarcinoma; the survival of tumor-bearing mice was prolonged and both metastatic foci and liver weight were decreased. These results suggest that PSK may be effective for the suppression of liver metastasis through activation of liver-associated NK cells.

Percutaneous endovascular embolization of intracerebral arteriovenous malformations. Experience in 72 cases.

Endovascular therapeutic embolization of arteriovenous malformations (AVMs) of the brain was performed in 72 patients between October 1986 and March 1993. From October 1986 to May 1991, 38 patients in this series were treated with isobuty1-2-cyanoacrylate (IBCA) glue. After June 1991, in the remaining 34 patients the embolic materials used included surgical silk (5-0), polyvinyl alcohol (PVA), ethanol, and estrogen. After treatment, 29 patients (40.3%) had complete angiographic obliteration of AVM, 15 (21%) transient neurologic postembolization deficit, and 3 (4%) permanent deficit without death. Embolic agents and procedures are discussed as to their curative effects, safety, normal perfusion pressure breakthrough (NPPB) problem, with silk mixture fluid being considered preferable.

In vitro activity of ofloxacin against Mycobacterium tuberculosis.

For the past 3 years, ofloxacin has been widely used in treating patients with drug-resistant tuberculosis in Taiwan. To study its usefulness in treating these patients, 139 isolates of Mycobacterium tuberculosis from patients treated at the Taiwan Provincial Chronic Disease Control Bureau from September 1994 to September 1995 were tested to determine the in vitro antituberculosis activity of ofloxacin. Of these, 131 had not been previously exposed to ofloxacin, and 130 (99.2%) were susceptible to ofloxacin. Sixty-four isolates were found to be susceptible to all conventional antituberculosis drugs, and all of these were also susceptible to ofloxacin. Of the remaining 67 isolates that were resistant to one or more conventional antituberculosis drugs, 66 (98.5%) were susceptible to ofloxacin. There was no association between susceptibility to ofloxacin and susceptibility to conventional antituberculosis drugs among the isolates tested. Of the eight isolates of M. tuberculosis previously exposed to ofloxacin, seven (87.5%) were resistant. Our results indicate that patients with multidrug-resistant strains of M. tuberculosis who have not received prior ofloxacin treatment may be safely treated with ofloxacin even without knowing the result of pretreatment ofloxacin susceptibility tests. We also found that ofloxacin resistance emerges frequently. Therefore, an adequate combination of antituberculosis drugs, along with ofloxacin, should be prescribed to prevent the development of resistance to ofloxacin.

Occupational exposure and respiratory morbidity among asbestos workers in Taiwan.

To determine the prevalence of asbestos-related lung disease and the impairment of lung function among asbestos workers, we conducted a cross-sectional health survey of 459 workers in 33 asbestos-related factories in Taiwan. Each worker was asked about his medical and occupational history and was given a medical examination, chest roentgenogram and pulmonary function test. Manufacturing processes included production of asbestos cements, textiles, friction materials and insulation products. Exposure assessments were based on asbestos sampling and counting using a phase contrast microscope. The average age of the participants in the study was 41.6 years. They had an average of 8.1 years of dust exposure, with a range of one to 42 years. The majority had a cumulative asbestos exposure of less than 20 fiber years/mL. No case of asbestos-related lung disease was found during our investigation. No roentgenogram showed unequivocal changes of asbestosis. However, a multiple linear regression analysis of the pulmonary function test showed that both FVC and FEV1 decreased significantly with an increasing cumulative dose of exposure after controlling for age, height and smoking effects during analysis. FEV1/FVC and FEF25-75% were not affected by exposure dose. The absence of asbestosis and other asbestos-related lung diseases may be due to an inadequate induction time of asbestos exposure and a possible healthy selection of workers. We conclude that among workers in Taiwan, there is a significant effect on the respiratory system, especially pulmonary function, due to asbestos exposure.

Quantitative measurement of drug susceptibility of Mycobacterium tuberculosis for monitoring chemotherapy response.

Changes in the degree of drug susceptibility of the bacterial population in tuberculosis patients during chemotherapy can be monitored by testing Mycobacterium tuberculosis isolates against various concentrations of drugs in 7H12 broth to determine radiometrically the minimal inhibitory concentration (MIC). The MICs were determined for multiple cultures isolated from 13 patients, of whom nine responded to chemotherapy within a few months, and four failed to respond at all. No changes in MICs were observed with the isolates obtained from the patients with a favorable response to chemotherapy. Determination of the MICs indicated that the failure in two cases out of four was associated with a steady increase in the degree of resistance of the patients' bacterial population. In two other failure cases, lack of change in the MIC values for multiple cultures suggests that the failure was associated with factors other than drug resistance. During the period of chemotherapy, quantitation of the degree of drug susceptibility in terms of broth-determined MICs may help to identify promptly those patients whose bacterial population is becoming resistant to the administered antimicrobial agents. For those patients who do not respond to chemotherapy, lack of change in the degree of resistance of their bacterial population may alert the physician to attempt to identify causes other than drug resistance for the chemotherapy failure.

Annual risk of tuberculous infection in Taiwan, 1996-1998.

Tuberculosis is still an important public health issue in Taiwan, and monitoring the trend of annual risk of infection (ARI) with Mycobacterium tuberculosis is essential. In this study, we conducted tuberculin skin tests to estimate the prevalence and annual risk of M. tuberculosis infection in first-grade schoolchildren in Taiwan Province. Because mass bacille Calmette-Guérin (BCG) vaccination programs have been carried out here, only non-BCG-vaccinated students were tested. From September 1996 through June 1998, there were 520,866 registered first-grade elementary school students in Taiwan Province. Of them, 15,147 (2.9%) were non-BCG-vaccinated, as determined by the absence of a BCG scar. All of them were tested for M. tuberculosis infection with 1 tuberculin unit (0.1 mL injection) of purified protein derivative RT23, by means of the Mantoux technique. Among the tested schoolchildren, 430 (2.8%) had a positive tuberculin reaction. Thus, the calculated ARI was 0.44%. The ARI varied in different areas of Taiwan, being highest (1.04%) in Nantou County and lowest (0.14%) in Miaoli and Tainan Counties. The ARI in aboriginal areas (1.16%) was 2.7 times that in nonaboriginal areas (0.42%). Our results indicate that the M. tuberculosis ARI is still high in Taiwan. To achieve the World Health Organization target of less than 0.1% for industrialized countries, we must intensify tuberculosis control programs in Taiwan.

Morbidity and mortality trends of pulmonary tuberculosis in Taiwan.

An epidemiologic study of pulmonary tuberculosis was carried out to examine the secular trend and geographic variations of the incidence, prevalence and mortality of pulmonary tuberculosis in Taiwan. The prevalence and incidence rates declined-steadily from 1910 to 1980, but have remained unchanged during the last decade. Mortality also showed an 8.0% annual decrease from 1974 to 1986, but has remained constant without any apparent changes in recent years. Mortality for pulmonary tuberculosis is much higher in the aboriginal areas, where patients have a poor adherence to treatment and where a high prevalence of alcohol abuse exists. The mortality for pulmonary tuberculosis in Taiwan is higher than that in most developed countries, such as the U.K. and the U.S.A., but is lower than that in mainland China and the Republic of Korea.

Initial drug resistance of Mycobacterium tuberculosis in Taiwan.

The prevalence and mortality rate of pulmonary tuberculosis in adults are high in Taiwan. Because the emergence of drug-resistant tuberculosis is one of the major causes of this sustained high tuberculosis mortality, surveillance of initial drug resistance is important. We tested Mycobacterium tuberculosis isolates from 1,935 newly diagnosed tuberculosis patients from January 1990 through December 1995 at the Taiwan Provincial Chronic Disease Control Bureau. The overall initial drug resistance rate was 12.3%; 8.7% of isolates were resistant to only one drug, 2.6% to two drugs, 0.7% to three drugs, and 0.3% to four drugs. The resistance rates to individual drugs were: streptomycin, 5.7%; isoniazid, 9.2%; ethambutol, 0.7%; and rifampin, 1.5%. The frequency of multidrug-resistant M. tuberculosis (resistant to at least isoniazid and rifampin) was 1.2%. In view of the high initial isoniazid resistance rate and low initial ethambutol resistance rate, ethambutol should be added to the regimen for the initial treatment of tuberculosis in Taiwan. The emergence of multidrug-resistant M. tuberculosis is ominous and should be considered when treating patients in Taiwan.

Family factors affecting the outcome of tuberculosis treatment in Taiwan.

Despite the progress of tuberculosis (TB) control, the failure of treatment mainly due to non-adherence to medical recommendations remains a major obstacle to the eradication of this disease in both industrialized and developing countries. We interviewed 397 active pulmonary TB cases (274 males and 123 females) in Taiwan and followed up their treatment outcomes. No significant differences were found between any of the groups of either sex on TB cognition scores. Positive correlations between cognition scores and compliance scores, between cognition scores and family apgar scores, and between compliance scores and family apgar scores were noted in this study. In males, the complete treatment group had a higher compliance score (mean +/- standard deviation = 13.45 +/- 2.80) and family apgar score (22.44 +/- 2.29) than the incomplete treatment group (11.61 +/- 3.21 and 14.77 +/- 3.92, respectively). In females, the complete treatment group had a higher cognition score (65.99 +/- 6.75), compliance score (13.65 +/- 2.55) and family Apgar score (22.78 +/- 2.30) than the incomplete treatment group (62.05 +/- 6.91, 11.70 +/- 3.04 and 14.84 +/- 3.80, respectively). In the logistic regression analysis, both the family Apgar score and compliance score were significantly related to the TB treatment outcomes. The subjects with a family Apgar score higher than 20 had a chance to complete treatment as high as 72-fold (95% confidence interval = 31.7-164.0) that of those with a score less than 20.(ABSTRACT TRUNCATED AT 250 WORDS)