[Source Apportionment of Morphine in Wastewater].

It is a new approach to identify legal or illegal use of morphine through information on municipal wastewater. However, the sources of morphine in wastewater are complex, and distinguishing the contribution of different sources has become a key issue. A total of 262 influent samples from 61 representative wastewater treatment plants in a typical city were collected from October 2022 to March 2023. The concentrations of morphine, codeine, thebaine, papaverine, noscapine, and monoacetylmorphine were analyzed in wastewater and poppy straws. Combined with the proportion of alkaloids in poppy straws, the source analysis of alkaloids in wastewater was analyzed using the ratio method and positive matrix factorization model (PMF). Only five alkaloids were detected in wastewater, and monoacetylmorphine, a metabolite of heroin, was not detected. The concentrations of morphine and codeine were significantly higher than those of noscapine, papaverine, and thebaine. By constructing the ratios of codeine/(morphine + codeine) and noscapine/(noscapine + codeine), the source of poppy straw could be qualitatively distinguished. The PMF results showed that three sources of morphine for medical use, poppy straw, and codeine contributed 44.9%, 43.7%, and 9.4%, respectively. The different sources varied in these months due to the COVID-19 and influenza A outbreaks, in which the use of drugs containing poppy straws and codeine was the main source, whereas the use of morphine analgesics remained relatively stable. Inventory analysis further demonstrated the reliability of the source contributions from the PMF model, and morphine was not abused in this city.

Taste traces: Capsaicin and sweeteners as anthropogenic markers in municipal wastewater.

Dietary-derived substances possess significant potential as anthropogenic markers owing to the large consumption and different intake habit. To investigate and evaluate such markers, wastewater samples from 35 wastewater treatment plants across 29 Chinese cities were collected to analyze artificial sweeteners (acesulfame and cyclamate) and natural spicy compounds (capsaicin and dihydrocapsaicin). Acesulfame (mean: 14.6 µg/L), cyclamate (mean: 24.3 µg/L), and capsaicin (mean: 101 ng/L) can be further investigated as anthropogenic markers due to their high detection frequency at high concentrations. Spatial use patterns revealed that acesulfame (5.31 g/d/1000 inhabitants (inh)) and cyclamate (8.16 g/d/1000 inh) use in northern China notably surpassed that in southern China (1.79 g/d/1000 inh and 3.23 g/d/1000 inh, p < 0.05). Conversely, chili pepper use was significantly higher (p < 0.05) in southern China (6702 g/d/1000 inh) than in northern China (2751 g/d/1000 inh), signifying a preference for sweetness in the northern regions and a predilection for spiciness in the southern regions. The total annual use of acesulfame (1842 t), cyclamate (3110 t), and chili (18.4 million tonnes) in China was estimated by this study, which was close to the national statistical production. In addition, sweetener use was negatively associated with the elderly population ratio, suggesting that the elderly population might not consume sweet foods. This study reveals the dietary sources of anthropogenic markers, highlighting the need for further research on the environmental implications of such markers.

Surveillance of COVID-19 and influenza A(H1N1) prevalence in China via medicine-based wastewater biomarkers.

The simultaneous monitoring of individual or multiple diseases can be achieved by selecting therapeutic medicines used to treat the primary symptoms of the condition as biomarkers in wastewater. This study proposes a novel approach to monitor the prevalence of COVID-19 and influenza A (H1N1) by selecting nine medicines to serve as biomarkers, including three antipyretics, three antivirals, and three cough suppressants. To verify our approach, wastewater samples were collected from seventeen urban and five rural wastewater treatment plants (WWTPs) in a Chinese city over a period of one year. The use of antipyretics increased notably during the COVID-19 pandemic, while the consumption of antivirals for influenza A (H1N1) rose in the post-COVID-19 pandemic period, indicating a minor spike in the occurrence of influenza A (H1N1) after the COVID-19 pandemic. Fever is a significant symptom of COVID-19 and can serve as a reliable indicator of disease prevalence. Our research found that the prevalence of COVID-19 in urban areas was significantly higher (at 78.5 %, 95 % CI: 73.4 % - 83.9 %) than in rural areas (with a prevalence of 48.1 %, 95 % CI: 42.4 % - 53.8 %). The prevalence of COVID-19 in urban areas in this study was consistent with the data reported by the Chinese center for Disease Control and Prevention (82.4 %). Continuous monitoring of WWTPs in urban areas with fluctuating populations and complex demographics can provide early disease warning. Our results demonstrate the feasibility of evaluating community disease prevalence by selecting major therapeutic medicines as biomarkers in wastewater.

Evaluation of eight psychoactive drugs used in Chinese cities by wastewater-based epidemiology.

With rapid economic development, an increasing number of people suffer from mental health diseases, which are gradually receiving the attention of society. However, basic data from surveys of mental disorders are limited. Composite influent samples were collected from 26 wastewater treatment plants in 23 major cities in China. The concentrations of the psychoactive drugs diphenhydramine, fluoxetine, doxepin, imipramine, sulpiride, zolpidem, carbamazepine, and flunitrazepam in the wastewater were determined. The detection frequency of diphenhydramine, sulpiride, and carbamazepine was close to 100 %, whereas that of the compounds was lower than 35 %. Carbamazepine had the highest mean consumption (31.1 mg/d/1000 people), followed by diphenhydramine (10.4 mg/d/1000 people) and sulpiride (11.3 mg/d/1000 people). Wastewater-based epidemiology (WBE) estimates of the average use of the three drugs were lower than those from the drug statistics data. Consumption of diphenhydramine in northern China was higher than that in southern China. A correlation analysis of psychotropic and illicit drugs revealed a correlation between sulpiride and heroin use, which may be related to the adverse effects of sulpiride treatment after heroin withdrawal. Psychotropic drug use is associated with both economic and social factors. We found associations between the use of the three drugs and age, occupation, and obesity, which are risk factors for mental disorders. The results showed that the monitoring of psychotropic drug using WBE has a certain reference value for public health care and for improving the understanding of mental disorders.

Assessment of correlations between sildenafil use and comorbidities and lifestyle factors using wastewater-based epidemiology.

Sildenafil (SIL) is widely used to treat erectile dysfunction. Information on its consumption and the factors influencing its use is limited in China. In this study, we sampled composite influent wastewater samples from 33 Chinese cities and analyzed SIL using liquid chromatography-tandem mass spectrometry. SIL consumption was estimated using wastewater-based epidemiology (WBE) and ranged from 10.6 mg/d/1000 people to 132 mg/d/1000 people, with a mean of 53 mg/d/1000 people. Prescription sales (3570 kg) accounted for 13.3% of the estimated SIL use (26842 kg) in 2018, thereby implying that SIL illicit use was greater than prescription use in China. Some regional differences were observed in SIL use, which was significantly higher in North China than South China (p < 0.05), thereby reflecting that the prevalence of SIL was affected by differences in lifestyle and socioeconomic factors. We found significant positive correlations between SIL use and consumption of allopurinol, hydrochlorothiazide, nicotine, and alcohol, thereby suggesting that the prevalence of SIL was associated with the prevalence of gout, hypertension, smoking, and drinking. Moreover, age structures, internet use, and marriage rates were positively correlated with SIL use, whereas the unemployment rate was negatively correlated with SIL use. Our study demonstrates that WBE is valuable for medical research to investigate licit and illicit drug use and to assess the underlying associations of different chemical uses.

Presence of the ketamine analog of 2-fluorodeschloroketamine residues in wastewater.

Ketamine (KET) analogs are increasingly emerging as new psychoactive substances (NPS). The present report describes the first detection of the KET analog, 2-fluorodeschloroketamine (2F-DCK), in influent samples collected from nine wastewater treatment plants in seven major Chinese cities from 2018 to 2020 by wastewater-based epidemiology (WBE). An analytical method based on solid-phase extraction and subsequent gas chromatography-mass spectrometry was developed for the detection of 2F-DCK and KET. The stability experiments showed that 2F-DCK and KET remained stable in wastewater for 15 days at room and frozen temperatures, and at two pH values (pH = 7 and pH = 2), with residue amounts between 90% and 110%. KET was detected in all samples, whereas 2F-DCK was detected in only four samples: from Guangzhou in 2018, Shenzhen in 2019, and Quanzhou and Nanning in 2020, indicating that 2F-DCK has been used as early as 2018 in China. The renal clearance of 2F-DCK was predicted based on the quantitative structure-pharmacokinetic relationship model, which was used to calculate an excretion factor of 3.7. The 2F-DCK consumption in four cities ranged from 3.71 ± 0.05 to 55 ± 0.09 mg/day/1000 inh, and KET ranged from 1.3 ± 0.04 to 76.5 ± 4.63 mg/day/1000 inh. This is the first study to investigate 2F-DCK by WBE, which provides relevant real-time data on the growth of NPS use, as well as useful information for the government to develop new policies.

Estimating dynamic population served by wastewater treatment plants using location-based services data.

Wastewater-based epidemiology is a useful approach to estimate population-level exposure to a wide range of substances (e.g., drugs, chemicals, biological agents) by wastewater analysis. An important uncertainty in population normalized loads generated is related to the size and variability of the actual population served by wastewater treatment plants (WWTPs). Here, we built a population model using location-based services (LBS) data to estimate dynamic consumption of illicit drugs. First, the LBS data from Tencent Location Big Data and resident population were used to train a linear population model for estimating population (r2 = 0.92). Then, the spatiotemporal accuracy of the population model was validated. In terms of temporal accuracy, we compared the model-based population with the time-aligned ammonia nitrogen (NH4-N) population within the WWTP of SEG, showing a mean squared error of < 10%. In terms of spatial accuracy, we estimated the model-based population of 42 WWTPs in Dalian and compared it with the NH4-N and design population, indicating good consistency overall (5% less than NH4-N and 4% less than design). Furthermore, methamphetamine consumption and prevalence based on the model were calculated with an average of 111 mg/day/1000 inhabitants and 0.24%, respectively, and dynamically displayed on a visualization system for real-time monitoring. Our study provided a dynamic and accurate population for estimating the population-level use of illicit drugs, much improving the temporal and spatial trend analysis of drug use. Furthermore, accurate information on drug use could be used to assess population health risks in a community.

Wastewater analysis reveals urban, suburban, and rural spatial patterns of illicit drug use in Dalian, China.

Illicit drug use in rural and suburban areas of China has not been studied extensively, as most studies have focused on illicit drug use in urban areas. To compare the differences between urban, suburban, and rural drug use, we collected influent samples from 19 urban, 9 suburban, and 18 rural wastewater treatment plants in Dalian, respectively. A method using solid-phase extraction combined with derivatization for gas chromatography -mass spectrometry analysis was applied to detect biomarker concentrations. The concentrations of methamphetamine and morphine ranged from 3.12 to 605 ng/L and < 2.35 to 198 ng/L, respectively. Norketamine was found in only four samples (5.56 to 14.5 ng/L), while 3,4-methylenedioxymethamphetamine and benzoylecgonine were not detected in any samples. Methamphetamine use in rural areas (16.3 mg/day/1000 inhabitant (inh), prevalence: 0.06%) was significantly lower than those in urban (77.1 mg/day/1000 inh, prevalence: 0.23%) and suburban (234 mg/day/1000 inh, prevalence: 0.70%) areas. Heroin use in suburban areas (57.6 mg/day/1000 inh, prevalence: 0.10%) was significantly higher than that in urban (13.9 mg/day/1000 inh, prevalence: 0.02%) and rural (8.68 mg/day/1000 inh, prevalence: 0.02%) areas. The results indicate relatively low levels of illicit drug use in rural areas of Dalian, related to low incomes and outflow of the working-age population. Illicit drug use was most prevalent in suburban areas of Dalian, which may be influenced by large floating populations and few anti-drug efforts in suburban areas.

Tracing consumption patterns of stimulants, opioids, and ketamine in China by wastewater-based epidemiology.

Illicit drug use has long been a key issue of international concern, and the true situation is unknown to the relevant authorities. To develop a profile of comprehensive consumption patterns of illicit drugs in China, data from 34 wastewater treatment plants in 25 cities were collected to analyze four classes of drugs, including amphetamine-type stimulants, opioids, ketamine, and cocaine. They were identified and quantified in samples using methods based on gas chromatography coupled to mass spectrometry. According to the wastewater-based epidemiology (WBE) approach, an analysis of the consumption pattern was performed regarding per inhabitant consumption based on the revised metabolic rate. The consumption quantity of illicit drug and precursor was divided into four categories based on statistical difference analyses: methamphetamine and ephedrine (precursor) were the predominant drugs in the first category, followed by ketamine and heroin in the second category, methcathinone and 3,4-methylenedioxymethamphetamine (MDMA) in the third category, and cocaine and methadone in the fourth category. There were distinctive spatial patterns: heroin and cocaine consumption was higher in Southern China than in Northern China, heroin consumption was higher in Western China than in Eastern China, and the consumption of each drug differed across seven regions of China, especially with ephedrine and methcathinone consumption higher in North China; heroin consumption higher in Southwest, Central, and Northwest China; and ketamine and MDMA consumption higher in East, South, and Central China. Compared with findings in previous studies, there were temporal patterns, in which ketamine consumption presented a downward trend but heroin remained stable. Based on correlation analyses, there were the polydrug abuse patterns between heroin and cocaine, methcathinone and ketamine, and cocaine and MDMA. In general, this study based on WBE provides a comprehensive evaluation of drug consumption in China.

Spatial analysis of metformin use compared with nicotine and caffeine consumption through wastewater-based epidemiology in China.

Monitoring the consumption of pharmaceuticals and licit drugs is important for assessing the needs of public health owing to the impact on individuals as well as society. The present work applied wastewater-based epidemiology to profile the spatial patterns of metformin, nicotine, and caffeine use and their correlations. Influent wastewater samples were collected from 27 wastewater treatment plants in 22 typical Chinese cities that covered all geographic regions of the country. The consumption of metformin ranged from 0.02 g/d/1000 inh to 8.92 g/d/1000 inh, whereas caffeine and nicotine consumption ranged from 4.33 g/d/1000 inh to 394 g/d/1000 inh and 0.17 g/d/1000 inh to 1.88 g/d/1000 inh, respectively. There were significant regional differences in the consumption of caffeine, with the highest consumption in East China and the lowest consumption in Northeast China. The consumption and concentration of caffeine were related to the gross domestic product and per capita disposable income of urban residents, respectively. There was a correlation between the concentrations of caffeine and cotinine (a nicotine metabolite), thereby indicating that individuals that use one of these substances are likely to use the other substance. A significant relationship was found between the concentration of metformin and cotinine, thereby implying that the use of tobacco may be correlated with type 2 diabetes. Co-analysis of these substances in wastewater may provide a more accurate picture of substance use situations within different communities and provide more information on human health, human behavior, and the economy. This report describes the newest study related to the consumption of metformin among the general population in China.

Assessment of metformin, nicotine, caffeine, and methamphetamine use during Chinese public holidays.

Metformin, nicotine, caffeine, and methamphetamine are widely used in China. However, the consumption pattern of these substances among the general population during Chinese public holidays may be different. Influent wastewater samples were collected from a municipal wastewater treatment plant in Yingkou, China on public holidays (n = 6) and working days (n = 20) to examine the changes in metformin, nicotine, caffeine, and methamphetamine consumption. The consumption of metformin in the city ranged from 1.94 to 14.4 g d-1 1000 people-1, while that of nicotine, caffeine, and methamphetamine ranged from 0.46 to 2.18 g d-1 1000 people-1, 31.8-89.8 g d-1 1000 people-1, and 87.0-657 mg d-1 1000 people-1, respectively. The highest consumption of metformin, caffeine, and methamphetamine was observed during the Spring Festival. The results indicated that the consumption of these substances showed almost the same trend and was higher during the Spring Festival, which may be influenced by personal habits and traditional festival reunions. There is a correlation between the consumption of nicotine and caffeine. Moreover, there are correlations between the consumption of metformin and nicotine, caffeine, as well as methamphetamine. The information based on wastewater epidemiology and consumption behavior, suggests that drug use in the region requires more attention or monitoring during holidays.

Methamphetamine use in typical Chinese cities evaluated by wastewater-based epidemiology.

Methamphetamine has become one of the most widely used illicit drugs in China. To understand the current situation in China, the prevalence and consumption of methamphetamine were estimated through wastewater-based epidemiology (WBE) in the present study. Methamphetamine concentrations ranged from 42.6 ng/L (Harbin) to 700 ng/L (Xi'an) in influent wastewater samples collected from 27 wastewater treatment plants (WWTPs) in 22 Chinese cities. The estimated consumption of methamphetamine was 23.0 (Dingxi) to 376 (Xi'an) mg/day/1000 inhabitants with a mean value of 157 mg/day/1000 inhabitants. The annual consumption in 2018 was estimated to be 84 tons (95% confidence interval, 44-136), which was 26% lower than that in 2014. The prevalence of methamphetamine use was 0.64% (95% confidence interval, 0.18-1.25), indicating that more than five million people used methamphetamine in 2018. Although drug abuse is common in the country, the consumption showed a different spatial pattern, with the highest values in Central China and the lowest use in Northeast China, so drug use is still considered a geographic and culture-dependent behaviour. The results indicated that WBE can not only be used to assess the trends of illicit drug use, but also to analyse the spatial differences in the whole country, which will provide complementary evidence for the prevention and control of methamphetamine use.

Assessing the trend of diabetes mellitus by analyzing metformin as a biomarker in wastewater.

According to International Diabetes Federation estimates, China has the highest rate of diabetes in the world. To monitor the prevalence of diabetes mellitus (DM) in near real-time, a first-line medication for the treatment of type 2 diabetes, metformin, was used. Wastewater-based epidemiology (WBE) was applied to estimate the consumption of metformin in Dalian from 2015 to 2018. Quantification of metformin was undertaken using solid-phase extraction (SPE) and N-methyl-bis (trifluoroacetamide) derivatization prior to GC-MS analysis. The concentrations of metformin in eleven wastewater treatment plants (WWTPs) ranged from 1.7 µg/L to 239.0 µg/L, with an average value of 68.3 µg/L. For metformin consumption, there was a gradual increase from 12.1 mg/d/capita in 2015 to 28.4 mg/d/capita in 2018. Meanwhile, the prevalence of metformin in the Dalian population ranged from 1.6% in 2015 to 3.8% in 2018. Similarly, the prevalence of DM showed an increasing trend from 12.2% in 2015 to 21.6% in 2018, which is consistent with the data predicted by traditional surveys (15.2-19.8%). Additionally, the prevalence of DM in 2015 estimated based on WBE was 12.2%, which agreed with the results from the traditional survey (12.3%). These results indicated that the proposed method provided a feasible way to reveal the prevalence of DM through metformin monitoring by the WBE approach.

Wastewater analysis reveals spatial pattern in consumption of anti-diabetes drug metformin in China.

Metformin has been widely used as an oral drug for the treatment of diabetes mellitus. However, its consumption can be influenced by many economic and social factors. In this study, we investigated the spatial consumption pattern of metformin in China through wastewater-based epidemiology (WBE) approach. Influent wastewater samples were collected from 21 wastewater treatment plants (WWTPs) in 19 cities of the northeast China. A method using solid-phase extraction combined with N-Methyl-bis (trifluoroacetamide) derivatization for GC-MS detection was applied for metformin analysis. In 21 days, metformin showed high stability in wastewater at 24 °C and -20 °C. The mean concentrations of metformin in all WWTPs ranged from 2.42 µg L-1 to 53.6 µg L-1. The consumption of metformin was 0.66-15.6 mg d-1 capita-1 with the mean value of 5.54 ±â€¯4.28 mg d-1 capita-1. The prevalence of metformin ranged from 0.09% to 2.10% with an average of 0.74%. Both the consumption and prevalence of metformin displays significant spatial variations in northeast China. A statistical correlation analysis indicated that the consumption of metformin increases with the decrease of per capita disposable income of urban residents. To further predict the use of metformin in China, we developed a regress model and depicted a consumption map. The annual consumption of urban residents in Chinese provinces range from 1085-63,828 kg yr-1 with mean value of 25,347 kg yr-1, which would provide a certain reference value for public health care and diabetes control.

Reduction in methamphetamine consumption trends from 2015 to 2018 detected by wastewater-based epidemiology in Dalian, China.

BACKGROUND: Wastewater-based epidemiology (WBE) has become a useful tool in long-term or short-term continuous monitoring of illicit drugs consumption over the world. METHODS: We investigated the trend of methamphetamine (METH) use between 2015 and 2018 through WBE in Dalian, a typical Chinese city. Samples were collected in 11 municipal wastewater treatment plants (WWTPs). An analytical method, solid-phase extraction combined with trifluoroacetic anhydride derivatization prior to gas chromatography-mass spectrometry (GC-MS) analysis was applied to detect METH concentrations. RESULTS: During the sampling period, the METH concentrations increased slowly from 315 ± 243 ng/L in 2015 to 523 ± 549 ng/L in 2016, followed by a significant decrease with the concentrations 188 ± 187 ng/L in 2017 and 54.6 ± 42.9 ng/L in 2018. Ammonium nitrogen (NH4-N) was applied to estimate population size. The average coefficient of variation for population in 11 WWTPs was 35.3 ± 8.9%, reflecting the dynamic variations of population effectively. For METH consumption, there was a gradual increase from 2015 (231 mg/day/1000 people) to 2016 (414 mg/day/1000 people) and a significant linear decrease to 2017 (206 mg/day/1000 people) and 2018 (53.9 mg/day/1000 people). The prevalence of METH increased from 2015 (0.78%) to 2016 (1.06%), then decreased to 2017 (0.55%) and 2018 (0.17%), showed similar trends with the consumption. CONCLUSIONS: The obvious reduction trends of METH consumption via WBE over the period in Dalian provides objective evidence for declined METH consumption in local population. The reduction is probably due to the severe crack-down of illicit drugs by the government.

[Activated carbon catalyzed persulfate oxidation of azo dye acid orange 7 in aqueous solution].

Persulfate oxidation is a recently emerging advanced oxidation process for the pollution control. Persulfate (PS) degradation of azo dye acid orange 7 (AO7) in an aqueous solution was studied in the presence of suspended granular activated carbon (GAC) at ambient temperature (e. g., 25 degrees C). It was observed that in the GAC/PS combined system AO7 was not only a good decolorization as well as a good mineralization. The AO7 decolorization ratio, the removal efficiency of the naphthalene ring and the removal of TOC were 80%, 50% and 48%, respectively, at PS/AO7 molar ratio of 100: 1, GAC dosage of 1.0 g/L in 5 h reaction time. As a catalyst, GAC can be reused for several times. For each reuse cycle, AO7 removal efficiencies gradually decreased, however, with the oxidant concentration increased, the extent of the deactivation of GAC decreased. The removal efficiencies of AO7 for each reuse cycle hardly changed, and were all above 90% at PS/AO7 molar ratio of 500: 1, GAC dosage of 1.0 g/L in 5 h reaction time. The adsorption capacity of reused GAC decreased as the reuse time increased. The results of surface oxygen function groups, scanning electron microscopy (SEM) and energy disperse spectroscopy (EDS) analysis of reused GAC changed slightly. Therefore, the reason leading to deactivation of GAC could be that adsorption took up the active sites, thus reducing PS's opportunities to contact with the surface of GAC.

Construction of a recombinant adenovirus vector expressing IL-18BP/IL-4 fusion gene and the anti-inflammatory effect induced by this gene on lipopolysaccharide-stimulated synovial fibroblasts.

OBJECTIVE: To construct a recombinant adenovirus vector for expressing the IL-18 binding protein (IL-18BP)/IL-4 fusion gene and confirm the anti-inflammatory effect of this gene. MATERIALS AND METHODS: The recombinant virus expressing IL-18BP/IL-4 fusion protein (AD-IL-18BP/IL-4) was constructed. AD-IL-18BP/IL-4 was used to infect synovial fibroblasts (SF). ELISA and Western blot analysis were used to determine the expressions of the proteins IL-4 and IL-18BP. To investigate the protective effects of this vector on rheumatoid arthritis, SF were infected with AD-IL-18BP/IL-4 and stimulated by LPS (1 microg/ml) 4 h later. The expression levels of TNF-alpha, IL-6, IL-8, and IL-18 in the culture supernatant were detected by ELISA and production of PGE2 and NO was estimated. The protein expression of COX-2, iNOS, and NF-kappaB p50 in treated SF was analyzed by Western blot. RESULTS: AD-IL-18BP/IL-4 can effectively express the IL-18BP/IL-4 fusion protein. The expressions of TNF-alpha, IL-6, IL-8, and IL-18 were significantly inhibited in LPS-stimulated SF after treatment with AD-IL-18BP/IL-4. The production of PGE2 and NO was significantly decreased. Moreover, NF-kappaB p50, COX-2, and iNOS levels in SF were markedly suppressed by AD-IL-18BP/IL-4. CONCLUSION: AD-IL-18BP/IL-4 can suppress the production and expression of inflammatory cytokines such as COX-2, iNOS, and NF-kappaB in LPS-stimulated SF.

Sesquiterpene lactone parthenolide markedly enhances sensitivity of human A549 cells to low-dose oxaliplatin via inhibition of NF-kappaB activation and induction of apoptosis.

Aberrant activation of NF-kappaB has been proposed as the major cause of chemoresistance in lung cancer. Low-dose chemotherapeutic agents with limited toxicity and achieving profoundly enhanced efficacy by blocking NF-kappaB activation may be a useful strategy in cancer therapy. Thus, this study was performed to explore the effect of parthenolide, a natural NF-kappaB inhibitor, on human lung cancer A549 cells treated with low-dose oxaliplatin, as well as to determine the potential mechanisms involved. We incubated A549 cells with different concentrations of parthenolide in the absence or presence of a low-dose of oxaliplatin for 48 h. Then, cell proliferation was determined by MTT assay, and flow cytometry was used to study apoptosis. PGE(2) production in culture supernatants was detected by competitive ELISA, while expression of NF-kappaB/p65, COX-2, caspase-3 and caspase-9 proteins were analyzed by Western blot. Finally, compared to parthenolide or oxaliplatin alone, significant improvements in cell apoptosis and growth inhibition indexes were observed in the combined treatment. NF-kappaB/p65, COX-2, and PGE(2) expression were suppressed by the co-application; meanwhile, caspase-3 and caspase-9 proteins were obviously activated. These findings indicate that parthenolide could markedly enhance sensitivity of A549 cells to low-dose oxaliplatin by inhibiting NF-kappaB activation and inducing apoptosis. Parthenolide in combination with a low dose of oxaliplatin may be a beneficial chemotherapeutic strategy for patients who cannot tolerate the severe side effects of the drug at therapeutic concentrations.

[Cordyceps sinensis polysaccharide enhances apoptosis of HL-60 cells induced by triptolide].

OBJECTIVE: To investigate the effects of polysaccharide fraction of Cordyceps sinensis (PSCS) on triptolide (TPL)-induced apoptosis in the HL-60 cells and the involved molecular mechanism. METHODS: The cultured leukemia HL-60 cells were divided into three groups: control group, TPL group (cells were treated with 5 ng/ml TPL only), and PSCS+TPL cells group (cells treated with 5 ng/ml TPL and 100 microg/ml or 200 microg/ml PSCS for 18 h). Cell viability was tested by MTT assay and apoptotic cells were quantitatively measured by flow cytometry with Annexin V/PI double stain.The expressions of Caspase-3, 6, 7, 9 and NF-kappa B proteins were tested by Western blot. RESULT: MTT assay showed that different concentrations of PSCS inhibited the cell viability. Flow cytometry indicated that TPL markedly increased the apoptosis rate of the HL-60 cells, and PSCS enhanced the apoptosis in a dose-dependent manner. Western blot showed that TPL did not inhibit the expression of the Caspase-3, 6, 7, 9 and NF-kappa B proteins, and when cells were treated with PSCS, the expression of proteins decreased with the PSCS concentration rising. CONCLUSION: PSCS can enhance TPL-induced apoptosis in HL-60 cells and inhibit the expression of NF-kappa B and Caspase 3,6,7,9,which might be the possible signaling pathway of inducing apoptosis.

Expression of interleukin-18, IL-18BP, and IL-18R in serum, synovial fluid, and synovial tissue in patients with rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic immunological disease, the invasive monocytes/macrophages and lymphocytes present in synovial cells and synovial tissue produce many cytokines and inflammatory mediators by paracrine signaling and plays a role in the pathological progress in RA patients. Interleukin-18 (IL-18) is a representative proinflammatory factor and displays multiple biological functions. This study was designed to investigate the expression of IL-18 and its receptor (IL-18R) and IL-18 binding protein (IL-18BP) in serum, synovial fluid, and synovial tissue of patients with RA, and to identify the pathological role of IL-18 in RA. Serum, synovial fluid, and synovial tissue were obtained from RA patients. Samples from patients with osteoarthritis and healthy people were obtained as controls. Levels of IL-18, IL-18BP, and PGE2 in serum and synovial fluid were measured by enzyme-linked immunosorbent assay. The biological activity of IL-18 in serum and synovial fluid was detected on the basis of IFN-gamma secretion from IL-18-responding human myelomonocytic KG-1 cells. NO in serum and synovial fluid was detected by Griess reaction. Expression of IL-18, IL-18BP, IL-18R, iNOS, and COX-2 mRNA and protein in synovial tissues was determined by quantitative reverse transcriptase polymerase chain reaction and Western blot. This study shows the expression levels of IL-18, IL-18R, iNOS, COX-2, and the biological activity of IL-18 in both serum and synovial fluid and tissue of patients with RA were significantly increased compared with the corresponding samples from the two control groups. In addition, expression of IL-18BP in patients with RA was decreased compared with samples from the two control groups. In conclusion, the overexpression of IL-18 and IL-18R may play an important role in the pathogenesis of RA.