This ensures that the total mortality for any geographic area and gender is the same as Morris et al. [14], while maintaining an estimated distribution across wealth quintiles based on individual risk factors and quantitative relative risk estimates from the literature. Rotavirus mortality burden is estimated as deaths per 1000 live births. equation(2) RVBurdenr,q,s=RVMortr,s⋅RVRiskIndexr,q,sRVRiskIndexr,s All subpopulation means were calculated using appropriate sample weights

CH5424802 solubility dmso based on the design of each survey. Mortality risk was converted into Disability Adjusted Life Years (DALYs) based on standard methods using age weighting and discounting [27] and [28]. Previous studies have shown that over 98% of DALYs associated with rotavirus diarrhea in low income settings are associated with mortality [29] and [30], as a result we have not estimated DALYs associated with morbidity from acute cases. We estimated SCH727965 timing of projected deaths by combining overall rotavirus mortality estimates for each subpopulation and the estimated age distribution of events from Morris

et al. [14], combined with additional data from Clark and Sanderson [31] and [32]. Monthly rates were estimated for the first year of life, and annually for the subsequent 4 years of life. For any subpopulation and period t, mortality burden is estimated in Equation (3), as: equation(3) RVBurdenr,q,s,t=RVTimet⋅RVBurdenr,q,sRVBurdenr,q,s,t=RVTimet⋅RVBurdenr,q,swhere RVTimet is the fraction of deaths occurring in time period t. We estimated the coverage of a ‘generalized’ 3-dose rotavirus vaccine that would be delivered alongside DPT1–3 through a routine immunization program. Vaccine effectiveness was estimated for each subpopulation based on estimated coverage of each of three doses, the expected timing of receiving each dose, and expected efficacy of each dose over time. Vaccination coverage was estimated by geographic area, gender and wealth quintile. Dipeptidyl peptidase Due to concerted national and state efforts, coverage of routine vaccinations in India is

rapidly improving. We used three alternative sources to estimate coverage: 2005–2006 NFHS-3 [24], 2007–2008 District Level Health Survey (DLHS-3) [33], and the 2009 Coverage Evaluation Survey (CES) [34]. A fourth survey, the Annual Health Survey [35], [36] and [37], was also consulted but it does not provide national estimates and was used descriptively. For the NFHS and the DLHS3, we estimate coverage of DPT1, DPT2 and DPT3 for each geographic area r, sex s and wealth quintile q sub-population. Vaccination timing was estimated for all three doses using vaccination data for 1-year-olds from DLHS-3. Specifically, for each subpopulation we estimated the proportion of children receiving each dose by the end of each time period t.

Acute toxicity refers to harmful effects caused by high concentrations of aluminium. Descriptions are available particularly www.selleckchem.com/products/AZD2281(Olaparib).html with regard to dementia: The first description of the aluminium-related dementias can be traced back into the 1970s [23] and [24] and most studies report a positive link between aluminium accumulation and cognitive impairments. However, some study designs are highly variable and their quality is questionable. More recently, evidence has demonstrated that high aluminium exposure from, i.e., drinking water can trigger acute episodes of dementia in patients with renal insufficiency, providing strong evidence for the causal relationship with aluminium [25]. The use of silicic

acid has also been suggested to have a protective affect against the development of dementia [26], [27] and [28]. As previously mentioned, the bioavailability of aluminium in drinking water is, for instance, co-dependent on its silica content: large amounts of silicic acid in drinking water reduce the uptake of aluminium and vice versa [6] and [10]. Exley and co-workers [26] have demonstrated that

regular consumption of silicon-rich mineral waters reduce gastrointestinal uptake of aluminium and removal of systemic aluminium from the body. As a result, this buy GPCR Compound Library may provide the basis of a non-invasive means for a therapy to treat the symptoms of Alzheimer’s disease, in an attempt to reduce their body burden of aluminium. However, in-depth follow up studies involved in identifying clinical improvement of symptoms are at an early stage. In the 1940s, inhalation of aluminium was propagated as prophylaxis against silicosis in mine workers [29]. Examinations of these mine workers conducted in the study revealed the neurotoxic until effects of this aluminium

exposure [30]. In 1988, the drinking water of the Camelford community in Cornwall, UK, was accidentally contaminated with 20 t of aluminium sulphate. Follow-up examination in the affected population demonstrated the consecutive neurotoxic effects of aluminium [31]. In another study, a neuropathological examination of an exposed individual who died from an unspecified neurological condition was performed. High aluminium levels were measured in affected regions of the cortex, where a rare form of β amyloid angiopathy was identified [32]. Chronic toxicity refers to the harmful effects of protracted low-dose contamination. Increased concentrations of aluminium have been demonstrated in senile plaques in the brains of Alzheimer patients. The property of aluminium to produce amyloid-beta and cause damage to neurons, as well as epidemiologic connections, have been indicative of a relationship between aluminium and Alzheimer’s disease for decades. Current reviews cite respective, but sometimes contradictory, studies [33]. To summarise the current state of knowledge, Bondy et al.

Infected pigs may therefore become a source of infection for humans, even if the virus would not succeed in becoming endemic in the pig population. Humans in contact with high concentrations of infected pigs may be exposed to much higher amounts of virus than when exposed to infected humans. This could result in much more severe clinical symptoms, even in a higher mortality. Possible contact persons are not just the farmers and their family, but also include veterinarians, pig consultants, traders, transporters, visitors of pig markets and slaughterhouse personnel. A way to decrease the risk for people involved may be vaccination of pigs, with the primary aim of reducing virus excretion and therefore exposure of humans

to the virus. Conventional vaccines consist of whole viruses propagated in either embryonated chicken eggs or cell cultures, which are subsequently inactivated and adjuvanted. In case new such vaccines, based on new influenza check details subtypes, are needed, the development, registration and subsequent production takes a relatively long time, taking care of safety, efficacy and production issues. As an alternative a recombinant purified hemagglutinin (HA) could be used as a vaccine. One such recombinant, a secretable, soluble 3-Methyladenine chemical structure trimer of the HA ectodomain from the H1N1v influenza strain, was constructed and formulated

as a vaccine to be tested in swine. The aim of this study was to determine to what extent this vaccine is able to protect against infection with the H1N1v influenza strain, especially with respect to reducing virus replication and excretion. It was shown that the HA trimer was almost complete able to prevent virus replication and excretion Linifanib (ABT-869) after a double vaccination. The study was carried out with 18 pigs, divided into two groups of 9. In one group the pigs were vaccinated twice, with a four week interval. At the age of 10 weeks they were vaccinated for the first time. The other group was an unvaccinated control group. Three weeks after the second vaccination the animals in both groups were challenged, resp. inoculated with the H1N1v virus. At days 1 and 3 post inoculation

(p.i.) 3 pigs from each group were euthanized. The remaining 3 pigs in each group were euthanized at day 21 p.i., the end of the experiment. The design of the experiment was evaluated and approved by the Ethical Committee for Animal Experiments of the Animal Sciences Group. Nine-week-old piglets were purchased from a high-health breeding herd in which no seroconversions against any influenza subtype had been observed for more than 2 years. Before purchasing the pigs, all were tested individually with an NP-ELISA (IDEXX) and in hemagglutination inhibition assays against H1N1, H1N2 and H3N2 influenza virus strains that are endemic in the swine population. Based on H3 numbering, a cDNA clone corresponding to residues 16–524 of the HA from A/California/04/2009(H1N1) (Genbank accession no. ABW90137.

A concern with this trial, however, is the description of the control group as conventional therapy. The description of the activities includes mostly passive, non-goal directed movement; this would not be considered

typical by many therapists. At this stage in upper limb research there are proven interventions that find more can be used as comparison in order to determine a truly superior treatment. In this trial though the amount of time spent in therapy was equivalent, the repetition of the activities were not; if this had been comparable the conclusion of ‘more effective’ could be made. The conclusion is thus difficult to accept. There is mounting evidence that high repetitions of active, goal directed interventions are necessary for improved upper limb function and therefore need to be a key ingredient in conventional rehabilitation. “
“Summary of: Frobell RB, et al (2013) Treatment for acute anterior cruciate ligament tear: five year outcome of randomized trial. BMJ 346: f232. doi: 10.1136/bmj.f232. [Prepared by Nicholas Taylor, CAP

Co-ordinator.] Question: Doesearly Caspase inhibitor anterior ligament (ACL) reconstruction plus early rehabilitation improve outcomes 5 years after injury in patients with an ACL ligament tear compared with rehabilitation with the option of delayed surgery? Design: Randomised, controlled trial included blinded outcome assessment. Setting: Two hospitals in Sweden. Participants: Adults aged 18 to 36 years with an ACL tear not more than 4 weeks old to a previously uninjured knee were included. Key exclusion were playing professional sport, being less than moderately active, and having a full thickness meniscal lesion. Randomisation of 121 participants allocated 62 to the early ACL reconstruction group and 59 to a group having the option of delayed ACL reconstruction if needed. Interventions: Both groups received a similar rehabilitation program supervised

by physiotherapists in outpatient clinics with goals for attaining range of motion, muscle function, Tolmetin and functional performance. In addition, the intervention group had ACL reconstruction surgery within 10 weeks of injury. The comparison group with the option of delayed reconstruction had ACL reconstruction surgery when presenting with symptomatic knee instability. Outcome measures: The primary outcome was the change in the Knee Injury and Osteoarthritis Outcome score (KOOS) at 5 years. The KOOS comprises an overall score and 5 subscales (pain, symptoms, activities of daily living, sport and recreation, and knee related quality of life) scored from 0 to 100 with higher scores indicating better results. Secondary outcome measures included the short-form health survey (SF-36), the Tegner Activity Scale, and radiographic osteoarthritis. Results: 120 participants completed the study.

[1]) and assuming that vaccination does not affect duration of colonisation. The main learn more factor affecting how the bias in the estimated vaccine efficacy becomes negligible is the prevalence of colonisation at the time of vaccination. When the prevalence is close to 0 (left-hand panel), the mean of VEacq estimates from cross-sectional data closely approximate the true VEacq as long as the samples are collected

2–3 months after vaccination. When the prevalence of colonisation is higher (right-hand panel), the bias is initially clearly negative and becomes relatively small only after several months since vaccination. As a rule-of-thumb for both scenarios, the time from vaccination until nasopharyngeal RGFP966 research buy sampling is determined by the rate of clearance rather than the rate of pneumococcal acquisition. This is shown by comparison between the “high” vs. “moderate” scenarios for overall acquisition in Fig. 1. Under both scenarios, colonisation should be sampled

only after at least twice the average duration of a carriage episode has passed since the immune-response. In the example, the mean duration was approximately 2 months and the sampling should thus occur 4 months after the immuno-response or somewhat later. The results for the combined vaccine efficacy against acquisition and duration (VET) were similar (data not shown). Apart from the requirement of approximate steady-state at the time

of sampling, these there are other factors that rather favour early measurement of colonisation (e.g. the possibility of waning immunity or changes in exposure with age and/or season). In addition to bias, the precision of estimation and sample size (cf. Section 5) need to be considered. In general, the precision was poor in the first 2 months, in particular with low individual prevalence and moderate rate of pneumococcal acquisition (data not shown). Also serotype-specific estimates can be obtained from a cross-sectional study (cf. Section 4 in [1]). In general, their estimation performs similarly to the aggregate (i.e., all vaccine-type) efficacy. For serotypes with very low prevalence, however, the negative bias in the efficacy estimates is obviously somewhat bigger unless the sample size is very large. The sensitivity of detecting pneumococcal colonisation depends on the technique of specimen sampling and handling, and the methodology to culture, identify and serotype pneumococci [2]. The current standard, which is based on using a single nasopharyngeal swab to measure the prevalence of pneumococcal carriage, is simple and rapid. The sensitivity of a single swab to detect and identify the dominant pneumococcal serotype is high, being in the range of 85–100% [2], [3] and [4]. A key challenge to nasopharyngeal sampling remains the identification of multiple serotypes simultaneously colonising the nasopharynx.

The mixture was then poured into ice water (500 ml) and the separated solid product was collected by filtration, washed

with water, dried and crystallized from ethanol to afford compound 4. Yield: 65%. M.P: 239–240 °C. 1H NMR (DMSO-d6): δ 11.4 (s, 1H, NH), 7.9 (s, 1H, NH), 7.0–7.4 (m, 5H, SC6H5), 5.6 (s, 1H, C5H of pyrimidine). Anal Cacld for C10H8N2SO2: C, 54.54; selleckchem H, 3.63; N, 12.72. Found: C, 54.52; H, 3.62; N, 12.70. A mixture of 6-phenylthiouracil (4) (3 g, 0.0125 mol) and POCl3 (12.2 ml, 0.125 mol) was refluxed for 4–5 h. Excess of POCl3 was removed under reduced pressure and the mixture was treated with ice/water. The separated solid was extracted with ether (3 × 50 ml) and washed with 5% aq. sodium bicarbonate

solution (1 × 25 ml). Ether layer was collected and dried over anhydrous sodium sulfate. Evaporation of the solvent furnished the title compound 5. Yield: 72%. M.P: 48–50 °C. IR (cm−1): 749 & 705 (C–Cl). 1H NMR (DMSO-d6): δ 7.2–7.6 (m, 5H, SC6H5), 5.9 (s, 1H, C5H of pyrimidine). Mass: m/z = 257 (M+, 100%). Anal Cacld for C10H6N2SCl2: C, 46.91; H, 2.43; N, 10.94. Found: C, 46.45; H, 2.36; N, 10.60. To a solution of appropriate phenol (0.004 mol) in dry toluene (10 ml) was treated with 60% w/v sodium hydride (0.004 mol) in oil under an inert atmosphere. The mixture was warmed to 50–60 °C for 30 min to facilitate the formation of sodium salt. why After all the sodium hydride had reacted, the suspension www.selleckchem.com/products/CP-673451.html was cooled and a solution of 2,4-dichloro-6-(phenylthio)pyrimidine (5) (0.001 mol) in toluene

(10 ml) was added slowly at room temperature. After stirring the reaction mixture at 75–80 °C overnight, it was allowed to cool and the mixture was treated with water (25 ml). The separated solid was extracted with ether (3 × 25 ml) and washed with 10% aq. sodium hydroxide (3 × 25 ml). Ether layer was collected, dried over anhydrous sodium sulfate and evaporation of the solvent furnished the crude compounds, which were recrystallized from spirit yielded the title compounds 6a–g in 62–86% yield. Yield: 86%. M.P: 130–132 °C. 1H NMR (DMSO-d6): δ 7.0–7.5 (m, 15H, ArH), 5.9 (s, 1H, C5H of pyrimidine). Mass: molecular ion peak at m/z = 374 (M+, 100%). Anal Cacld for C22H16O2N2S: C, 70.96; H, 4.30; N, 7.52. Found: C, 70.89; H, 4.28; N, 7.50. Yield: 70%. M.P: 79–80 °C. 1H NMR (DMSO-d6): δ 6.8–7.5 (m, 13H, ArH), 5.9 (s, 1H, C5H of pyrimidine), 2.3 (s, 6H, CH3). Anal Cacld for C24H20O2N2S: C, 72.00; H, 5.00; N, 7.00. Found: C, 71.96; H, 4.97; N, 7.06. Yield: 63%. M.P: 80–82 °C 1H NMR (DMSO-d6): δ 6.9–7.5 (m, 13H, ArH), 6.4 (s, 1H, C5H of pyrimidine), 2.3 (s, 6H, CH3). Anal Cacld for C24H20O2N2S: C, 72.00; H, 5.00; N, 7.00. Found: C, 71.46; H, 4.96; N, 6.94. Yield: 68%.

We generated mouse monoclonal antibodies to determine B cell epitopes of the recombinant Hsp70 protein and focused on linear epitopes. Subsequently, epitope-specific antibody responses, induced by vaccination of cattle and goats with recombinant MAP Hsp70, were analyzed to assess whether these this website antibodies recognized the same linear epitopes. Lastly, the monoclonal antibodies were used to study if these antibodies recognized native MAP Hsp70 protein in lesional tissue in naturally infected animals and if they interact with intact bacteria. Two Balb/c mice,

obtained from Charles River (Someren, the Netherlands), were used for the generation of MAP Hsp70 specific monoclonal antibodies. Animals were kept under standard housing and care conditions at the Central Animal Facilities of Utrecht University (Utrecht, the Netherlands). Thirty female goat kids (Saanen breed dairy Galunisertib mw goats, age 14 ± 3 days at the start of the experiment) were used. The kids were raised using conventional procedures and feeds, and were checked daily for general health. They were randomly assigned to one of the four experimental groups. Goat kids in groups 1 (n = 7) and 2 (n = 8) (uninfected controls) were housed separately from goat kids in groups 3 (n = 7) and 4 (n = 8) (MAP infected). Goat kids assigned to groups 2 and 4 were immunized once at the start of the experiment (day 0). The immunization consisted of the administration of 200 μg of recombinant

MAP Hsp70 in 1 mL phosphate buffered saline (PBS) containing 10 mg/mL dimethyl dioctadecyl ammonium bromide (DDA) adjuvant (Sigma Aldrich, USA) in the final preparation, subcutaneously in the lower neck region. Goat kids assigned to groups 3 and 4 were infected orally with 3 oral doses, at days 0, 2 and 4, of 2 × 109 cfu of MAP strain G195, originally isolated from a goat with clinical signs of paratuberculosis, grown on Middlebrook 7H10 supplemented with OADC and Mycobactin J (a generous gift from D. Bakker, CVI, Lelystad, the Netherlands). The cfu of the infection dose was determined by colony counts of serial dilutions on 7H10 agar plates. Blood samples

were taken from the vena jugularis on a weekly basis for a period of 3 months. Serum was stored at −20 °C, until further use. Goats were euthanized at the end of the experiment Rebamipide and tissue samples from ileum, jejunum, the ileocecal and a jejunal mesenteric lymph node were analyzed using MAP specific IS900 PCR [16], bacterial culture on mycobactin J supplemented HEY medium (BD Biosciences, Belgium) and histopathology. Sera from cattle subjected to a Hsp70 vaccination – challenge experiment, published previously [9], were used to characterize MAP Hsp70 specific antibody responses. In short, 4 groups of 10 female calves aged 29 ± 9 days, randomly assigned to one of 4 experimental groups, were used in that study. Treatment of the groups was identical to the goat kids described in Section 2.1.3.

1%) blood samples and 21/50 (42.0%) CSF samples. As expected, CSF is the most suitable sample for diagnosis of meningococcal meningitis and blood is the most suitable sample in meningococcal sepsis. RT-PCR has always a greater sensitivity (2–8 times higher) when compared to culture, ranging from

2.3 times in the CSF of patients with meningitis, to 8.7 times in CSF of patients with sepsis. Over the study period there were 18 deaths, constituting an overall case fatality ratio (CFR) of 13.2%. Five out of 18 (27.8%) deaths occurred in the first year of age, 9 out of 18 (50.0%) occurred between the second and the fifth year of age; 3 cases occurred in adolescents (13–17 years of age). One case occurred at 6.2 years. CFR was 24.4% (11/45 cases) in children admitted with a diagnosis of sepsis, and 7.7% (7/91 cases) in children admitted for meningitis and in whom sepsis Olaparib clinical trial was not mentioned at admission. Twelve patients (8.9%)

had complications during the acute phase of disease (cutaneous or subcutaneous necrosis, acute renal failure, seizures). During the follow-up, severe sequelae selleck chemical such as abnormalities in Nuclear Magnetic Resonance of brain (gliosis, idrocephalus) associated with neurologic symptoms, mental retardation, amputation of both hand and foot fingers have been reported in 4 patients (3.0%). The results, obtained in a large pediatric population of Italian patients, demonstrate that invasive meningococcal infection has the highest incidence in the first 5 years of life where over 70% cases occur and in particular in the first year of age, where over 20% of all cases found in pediatric age are found. The incidence peak, similarly to what reported in other countries [16], is between the 4th and the 8th month of life. In parallel with the introduction of routine MenC vaccination in different Italian regions, the incidence of

meningococcal infection due to serogroup C has progressively decreased in infants and adolescents [8], [9], [13] and [17]. However, invasive meningococcal disease is still the first cause of meningitis and is second only to pneumococcal infection for cases of Chlormezanone sepsis. The most common cause of invasive meningococcal disease, accounting for over 80% of cases found in patients younger than 24 years of age [9] and [17] is now MenB. Culture has been, so far, the most used technique for meningococcal surveillance; however, bacterial culture leads to an important underestimation of disease burden. Confirming previous results, [16], [18] and [19] once again Realtime PCR results significantly more sensitive than culture in identifying meningococcal infection, independent of the biological sample used and the clinical presentation. In fact, in our data obtained in patient tested at the same time with both methods, sensitivity of culture was less than one third that of Realtime PCR.

It seems that the growing use of Kinesio Taping is due to massive marketing campaigns (such as the ones used during the London 2012 Olympic learn more Games) rather than high-quality, scientific evidence with clinically relevant outcomes. The widespread use of Kinesio Taping in musculoskeletal and sports physical therapy is probably further reinforced by the authors in some of the included trials concluding that Kinesio Taping was effective when their data did not identify significant benefits. Policymakers and clinicians should carefully consider the costs and the effectiveness of this intervention when deciding whether

to use this intervention. Although Kinesio Taping is widely used in clinical practice, the current evidence does not support the use of this intervention. However, the conclusions from this review are based on a number of underpowered studies. Therefore large and well-designed trials are greatly needed. The research group for this review is currently conducting two large randomised

controlled trials, which are investigating the use of Kinesio Taping in people with chronic low back pain; they should provide new and high-quality information on this topic. One of them31 click here compares different types of application of Kinesio Taping in 148 participants with non-specific chronic low back pain, with the outcomes of pain intensity, disability and global impression of recovery. The second trial32 tests the effectiveness of the addition of Kinesio Taping to conventional physical therapy treatment in 148 participants with chronic non-specific low back pain, with the outcomes of pain intensity, disability, global impression of recovery and satisfaction with care. It is expected that these two trials will contribute to a better understanding of this

intervention’s effectiveness. What is already known on this topic: Kinesio Tape is thinner and more elastic than conventional tape. Kinesio Taping involves application of the tape while applying tension to the tape and/or with the target muscle in a stretched position. Recent systematic reviews of trials of Kinesio Taping have identified insufficient, low-quality evidence about its effects, but new trials of Kinesio Taping are being Levetiracetam published frequently. What this study adds: When used for a range of musculoskeletal conditions, Kinesio Taping had no benefit over sham taping/placebo and active comparison therapies,the benefit was too small to be clinically worthwhile, or the trials were of low quality. Therefore, current evidence does not support the use of Kinesio Taping for musculoskeletal conditions. Some authors concluded that Kinesio Taping was effective when their data did not identify significant benefit. eAddenda: Figure 3 and Appendix 1 can be found online at doi:10.1016/j.jphys.2013.12.

8 ml/min was used. Detection was carried out at 220 nm. The injection volume was 20 μl; analysis was performed at ambient temperature. An accurately weighed quantity of miglitol (10 mg) was transferred to 10 ml volumetric flask and dissolved in water and diluted up to the mark with water to get a 1 mg/ml solution.

The series standard solutions were prepared by dilution of aliquots of the standard stock solution with mobile phase to get concentration in the range of 10–50 μg/ml of miglitol. Twenty microliter of the each standard solution was injected to HPLC system. The peak areas were plotted against the corresponding concentrations to obtain the calibration graph. The system suitability is used to verify whether the resolution and reproducibility of the chromatographic system are adequate for analysis to be done. The tests ABT-737 manufacturer were performed by collecting data from five replicate injections of standard solutions. A 20 μl standard drug solution was injected separately and system suitability parameters click here were recorded. Twenty tablets were weighed and average weight was calculated. The tablets were triturated to a fine powder. An accurately weighed quantity of powder equivalent to 10 mg of miglitol

was transferred to 50 ml volumetric flask. About 20 ml of water was added and sonicated for 15 min; further volume was made up to the mark with same solvent. The resulting solution was filtered and filtrate was appropriately diluted with mobile phase to get approximate conc. of 25 μg/ml of miglitol. Twenty micro liters of the test and standard solutions were injected separately after the equilibration of mobile phase with stationary phase. The chromatograms were recorded upto 8 min and area of each peak was noted. The optimized RP-HPLC method was completely validated according to the procedure described in ICH guidelines and United State Pharmacopoeia for validation of analytical methods. The performance parameters evaluated for the method were linearity, precision, accuracy, limits of detection and quantitation

and ruggedness. Linearity was studied by diluting standard stock solution at five Ergoloid different concentrations (n = 3) covering the range of 10–50 μg/ml for miglitol, respectively. A graph was plotted for the concentration of the corresponding drug versus peak area. The correlation coefficient (r2) for each drug was calculated. Repeatability study was carried out by analyzing sample solution six times, at 100% of test concentration within the same day using proposed method. Similarly, the intra and inter day precision was evaluated by analyzing tablet sample on the same day and on different days at different time interval, respectively. The contents of drugs and the % relative standard deviation (% R.S.D.) value were calculated.