Background: Considering the anti-oxidant properties of Pistacia atlantica and lack of data regarding its efficacy in the treatment of ulcerative colitis, this study aims at investigating the effect of the Pistacia atlantica fruit extract in treating experimentally induced colitis in a rat model.Methods: Seventy male Sprague-Dawley rats (weighing 220±20 g) were used. All rats fasted 24 hours before the experimental procedure. The rats were randomly divided into 7 groups, each containing 10 induced colitis with 2ml acetic acid (3%). Group 1 (Asacol), group 2 (base gel) and group 7 (without treatment) were assigned as control groups. Group 3 (300 mg/ml) and group 4 (600 mg/ml) received Pistacia atlantica fruit orally. Group 5 (10% gel) and group 6 (20% gel) received Pistacia atlantica in the form of gel as enema. Macroscopic, histopathological examination and MDA measurement were carried out.Results: All groups revealed significant macroscopic healing in comparison with group 7 (P<0.001). Regarding microscopic findings in the treatment groups compared with group 7, the latter group differed significantly with groups 1, 2, 4 and 6 (P<0.001). There was a significant statistical difference in MDA scores of the seven treatment groups =76.61, P<0.001). Post-hoc comparisons indicated that the mean±SD score of Asacol treated group (1.57±0.045) was not significantly different from groups 4 (1.62±0.024) and 6 (1.58±0.028).Conclusion: Our study showed that a high dose of Pistacia atlantica fruit oil extract, administered orally and rectally can improve colitis physiologically and pathologically in a rat model, and may be efficient for ulcerative colitis. Although a lower prevalence has been reported in Asia , IBD is on a rising trend.1-3 The exact cause of the disease is unknown, however few possible factors could be genetic, immunologic, environmental and oxidative stress.2,3 Additionally, evidence reveals that reactive oxygen species (ROS) and initiation of lipid peroxidation which are produced in inflamed mucosa, play an important role in the pathogenesis of colitis.4 Oxidative stress causes production of free radicals that elevate inflammatory mediators and damage intestinal mucosa.4,5 IBD patients also have low quality of life and risk factor for colon cancer.6Inflammatory bowel disease (IBD) is characterized by chronic, relapsing, idiopathic inflammation of the gastrointestinal tract. The two major forms of IBD are ulcerative colitis and Crohn’s disease.5 These drugs have various side effects; therefore, there is a rising trend in the world to use herbal drugs for treating ulcerative colitis.7 Herbal medicines are widely used in traditional methods of curing diseases among native people. Moreover, in recent years the therapeutic effects of some herbal drugs such as antioxidants are proven in clinical settings.4Anti-inflammatory agents such as glucocorticoids and salicylates as well as biological agents against tumor necrosis factor-α (TNF-α) are common treatments for ulcerative colitis.Pistacia atlantica or Bene (from the family Anacardiaceae) is a native fruit in Iran that is used in traditional herbal medicine.8Pistacia atlantica’s oil, which is called the Bene Hull Oil (BHO) has been recently introduced to the world as a highly stable compound with anti-oxidative properties. The components of BHO include 6.5% unsaponifiable matter (common in vegetable oil), carotenes, tocopherols and alcohols. Tocopherols and tocotrienols have antioxidant activity and act similar to vitamin E which is beneficial for human health.9,10Pistacia atlantica is believed to have several therapeutic properties such as relieving upper abdominal discomfort and pain, dyspepsia and peptic ulcer. Pistacia spp. is a diuretic and stimulant agents.11 Several studies have reported the antimicrobial and analgesic activity of Pistacia atlantica.12-14Pistacia atlantica might be an alternative medicine or beneficial food for the prevention or treatment of IBD patients. Considering the lack of data regarding its efficacy in treating ulcerative colitis, this study investigates the effect of the Pistacia atlantica fruit extract in treating experimentally induced colitis in a rat model.Antioxidant properties of Gel PreparationPistacia atlantica extracts  were added to deionized water. The mixtures were gradually added to the Na-CMC with glycerol and finally the prepared gel was homogenized for 30 minutes and all formulations were collected in an aluminum tube in the refrigerator.Gel preparation was done in a pharmacology lab under the supervision of expert technicians. The main substance was carboxymethyl cellulose (CMC). 2% sodium CMC was mixed in 5% glycerol and continuously stirred with a mixer at 500 RPM in order to prepare a gel-forming agent. In the next step, AnimalsThe present animal study was approved by the Ethics Committee of Shiraz University of Medical Sciences (registration number: 90-3894) and all relevant considerations regarding animal rights were taken into account. This study was carried out at the Laboratory Animal Center of Shiraz University of Medical Sciences, Shiraz, Iran. Seventy male Sprague-Dawley rats weighing 220±20g were used. Animals were housed in standard cages with free access to tap water and standard food (ad libitum) and maintained at a controlled temperature (23±1ºC) with a 12/12 hour light-dark cycle. All rats fasted for 24 hours before the experimental procedure. Extract PreparationPistacia atlantica fresh fruits were prepared from Shiraz  and authenticated by Pharmacology Department of Shiraz Medical School. For the preparation of oil extract, dried and finely powdered fruit (100 g) was soaked in an adequate volume of N-hexane (450 cc) and the extraction was continued for 48 hours to obtain the full extract using the percolation method. The extract was then filtered and evaporated in a rotary evaporator under reduced pressure resulting in a semisolid and oily extract. The final product was then prepared in two doses of 300 mg/kg and 600 mg/kg. The gel form was prepared in concentrations of 10% and 20%.Experimental DesignColitis was induced in the rats with 2 ml acetic acid 3% by intracolonic instillation through a polyethylene catheter, which was placed 8 cm from the anus under anesthesia with ketamine (90 mg/kg) and xylazine (5 mg/kg). The rats were randomly divided into 7 groups of ten, as follows:Groups 1 and 2 were the control groups, respectively receiving 1 ml Asacol  and 1 ml base gel (carboxymethylcellulose) as enema.Pistacia atlantica fruit oil extract orally, respectively.Groups 3 and 4 received 300 and 600 mg/kg Pistacia atlantica’s fruit as enema.Groups 5 and 6 received 10% and 20% gel from Group 7 was the third control group in which colitis were induced without treatment.2 chamber. The abdomen was opened and the colon was exposed. The distal 8 cm of the colon was removed and opened by longitudinal incision. The mucosal surface was washed with saline buffer and then the mucosal injury was macroscopically assessed using Morris et al.’s15 grading scale .All groups were subjected to relevant protocol and after 7 days, rats were sacrificed in a CO16 Inflammation and crypt damage was assessed and the sections were coded using a modified validated scoring scheme as described by Appleyard and Wallace.17For histological examination, the colon tissue was fixed in 10% formalin sectioned in 5 μm thick sections, and stained with haematoxylin and eosin. All slides were reviewed by a single blinded pathologist and sub-classified according to the “repair scoring system”.Tissue MDA Measurements18 MDA forms a colored complex in the presence of thiobarbituric acid (TBA) and n-butanol, which is detectable by the measurement of absorbance at 532 nm. Absorbance was measured using a Shimadzu UV-160 spectrophotometer. 1,1´,3,3´-tetramethoxypropane was used as standard and the results were expressed as µmol/g protein.19MDA production was measured in tissue samples by the method proposed by Ohkawa et al.Statistical AnalysisStatistical analyses were performed using SPSS 21 software . One way ANOVA, Kruskal-Wallis and Mann-Whitney tests were used. Data were expressed as mean±SD, frequency, and percentages. A P value <0.05 was considered as statistically significant. Moreover, Bonferroni correction was performed as an appropriate form of analysis. Macroscopic Findings15 Kruskal–Wallis test revealed significant macroscopic healing between group 7 (colitis induced without treatment) and other groups (P<0.001).All specimens were assessed according to the Moris scoring system.To determine the exact difference between the groups, Mann-Whitney U test between pairs of groups was performed. A statistically significant difference was seen between groups 7 and 1 (standard treatment with Asacol) (P<0.001), groups 7 and 4  (P=0.002) and groups 7 and 6 (extract 20% gel) (P=0.001). The Bonferroni correction method was applied according to the number of comparisons.Histopathological Findings16 With respect to the microscopic findings in the treatment groups compared with group 7 (colitis induced without treatment), as shown in Microscopic findings included focal crypt disorganization, ulceration, moderate colitis, and crypt disorganization, regeneration, granulation tissue and mild inflammation . The samMDA Test ComparisonA one-way between-group analysis of variance was used to analyze the impact of the new treatment on inflammation levels, as measured by the MDA test. There was a statistically significant difference in MDA scores of the seven groups =76.61, P<0.001). Post-hoc comparisons using the LSD test indicated that the mean±SD score for group 1 (1.57±0.045) was not significantly different from groups 4 (1.62±0.024) and 6 (1.58±0.028) as shown in Pistacia atlantica fruit extract as well as its effect on reducing inflammation and contribution to tissue repair was examined in acetic acid induced colitis in a rat model. Intrarectal acetic acid injection is an easy, repeatable, and cost-effective method for experimental colitis on small laboratory animals. It is comparable with inflammation and ulcers in the epithelium of the colon, and findings such as damaging the crypts of the intestine and killing epithelial cells and lowering goblet cells. These changes are similar to ulcerative colitis.19In this study, the effect of the Pistacia atlantica fruit oil extract reduces colonic injury by suppressing oxidative damage. The pathogenesis of the disease was assessed by evaluating different parameters such as macroscopic score, microscopic score and oxidative stress markers like MDA level. We found an improvement and lower MDA levels in the groups treated with the high dose of fruit extract compared with the untreated group. Histological and macroscopic data confirmed that both oral and rectal administration of Pistacia atlantica fruit oil extract could alleviate bowel inflammation. Asacole was used as the standard drug, which produced no significant difference in the above parameters in comparison with Pistacia atlantica at doses 600 mg/kg  and 20% gel form .The findings of this investigation demonstrated that Pistacia atlantica were found to be rich in proteins, oils, fibers, and unsaturated fatty acids, suggesting that they may be valuable as food. The major components of Pistacia atlantica are α-tocopherols and important sterols that have antioxidant properties and health benefits such as being precursors of vitamin D and lowering blood cholesterol.20 Their antioxidant activity is attributed to the presence of tocopherols and tocotrienols, and are as active as vitamin E.1,2 Shimizu et al.,21 showed that a vitamin E enriched diet could be beneficial in the treatment of ulcerative colitis in rats. The therapeutic role of Pistacia atlantica for ulcerative colitis in rats may be similar to vitamin E, which is consistent with findings by Shimizu et al.21The fruits of 22 found that vitamin E and selenium supplementation, significantly reduced both the severity of colonic lesions and the levels of MDA and protein carbonyl. The researchers offered antioxidants and some micronutrients as therapeutic agents. In another study, vitamin E levels were also found to be low in some patients with IBD and low levels of vitamin E was found during active disease.23Ademoglu et al.24,25 Vitamin E can also reduce intestinal inflammation by preventing the activation of the transcription factor NF-κB, which plays an important role in intestinal inflammation.26,27 This likelihood is supported by several in vitro studies in which vitamin E inhibited tumor necrosis factor-α-induced NF-κB activation in human Jurkat T-cells28 and inhibited the activation and translocation of NF-κB to the nucleus and the binding of the activated proteins to the κB DNA site.29Vitamin E has beneficial effects intraluminally. It is a lipid soluble radical scavenger and a lipid peroxidation suppressor, which reduces the free radical generating capacity of the feces in ulcerative colitis.4 found that Hypericum perforatum is effective in decreasing MDA levels in TNBS-induced colitis in rats due to its antioxidant activity. Consistently, in our study, MDA levels decreased in the treatment groups and this reduction may be attributed to the protective effect of Pistacia atlantica against lipid peroxidation, which may in turn be due to the presence of tochpherols and tocotrienols.Dost et al.30 CMC appears to be beneficial despite high levels of pro-inflammatory cytokines, suggesting a mechanism of action independent of an imbalance in cytokine production. Possible mechanisms could include an increase in cellulose derived nutrients to promote mucosal healing or immune function as well as variations in the intestinal flora. Thus, CMC may have exerted its positive effects on the intestinal lumen by increasing the viscosity of the mucous layer, thereby altering patterns of pathogenic bacteria migration through the mucous layer, where they would have otherwise led to inflammation and damage to the intestine.29In another study, CMC-treated mice had significantly lower histology scores and lower colon weight/length ratios, indicating that CMC-treatment alone may play a role in reducing colonic injury.Boswellia Serrata are effective in ulcerative colitis based on their antioxidant properties, which is similar to that of Pestacia atlantica.31 However, not all studies have shown consistent and promising results with respect to the role of herbal therapy in IBD. The reasons for such controversial findings could be related to different study designs, small sample size, lack of adequate control, and the variation in drug formulations and doses.31 All these confounding factors were considered in this study.Common belief indicates that herbal therapies are safer and less toxic than conventional treatment in patients with IBD. Herbal medicines such as Curcumin, Aloe Vera, and Pistacia atlantica, also has no proven hepatotoxicity effects.32,33 This treatment will be attractive to many patients and cost effective in comparison with glycocorticoid therapy. No toxic effect of Pistacia atlantica has been reported, showing that higher doses of this extract may be effective and safe. Recently, resistance to drugs, their toxicity and side effects have been reported; therefore, there is a great need for the development of an effective, safe and topical herbal drugs such as a Pistacia atlantica extract for different forms of ulcerative colitis.Most of the herbal therapies used for IBD have been associated with minimal adverse effects compared with glycocorticosteroids. According to previous studies, Pistacia atlantica exerts its beneficial effects and its efficacy should be confirmed through clinical trials. This study opens a path for future clinical trials in humans.This study had limitations such as the lack of evaluation of anti-inflammatory markers and stereology of colon tissue. Certainly, further studies are required to clarify the mechanisms by which Pistacia atlantica fruit extract orally and rectally can improve induced colitis physiologically and pathologically in a rat model. This therapy is potentially safe, cost-effective, and efficient for treating ulcerative colitis. Further investigations of its clinical application in human model are warranted.


The nationwide seroprevalence of hepatitis E IgG was determined among young men in Thailand. Overall seroprevalence was 14% (95% CI 13%–15%); range by province was 3%–26%. Seroprevalence was lowest in the south, an area predominantly occupied by persons of the Islam religion, whose dietary laws proscribe pork. Hepeviridae. HEV, identified in 1990, was a major cause of what was previously called non-A, non-B hepatitis (Hepatitis E virus (HEV), the etiologic agent of hepatitis E, is a nonenveloped, positive–sense single-stranded RNA virus, ≈7.2 kb in length, of the family In Thailand, hepatitis E outbreaks have not been reported; however, sporadic cases have been reported from many areas  during 2007–2008. The RTA uses a lottery system to select ≈60,000 young Thai men at the district level of their family residence for enlistment annually. The men enlisted comprise approximately 10% of young men at the district level in Thailand. Sample sizes were calculated to detect a seroprevalence of ≈50% in each province to within 10% of the true value with 95% confidence. A total of 7,760 stratified randomized samples were chosen on the basis of the reported province of residence of the men before RTA entry.2 2-tailed test; p value <0.05 was considered statistically significant. We performed the analyses using SPSS version 12 .We measured hepatitis E IgG antibody using a commercial IgG ELISA kit following the manufacturer's instructions . We tested associations between demographic characteristics and hepatitis E prevalence using the χThe study population is described in The hepatitis E IgG seroprevelence was lowest in the southern region. This region has the highest percentage of persons whose religion is Islam, known as Muslims, in Thailand. Because consumption of pork is proscribed by the religion of Islam, the region had the lowest pork consumption and pork production  (11).This study determined the nationwide seroprevalence of hepatitis E IgG in Thailand in young men from all 76 provinces. The overall hepatitis E seroprevalence at 14% (95% CI 13%–15%) is similar to results of previous hepatitis E seroprevalence studies in Thailand (9%–23%)  (In Thailand, reported swine HEVs belong to the G3 genotype and were genetically closely related to HEV isolates from patients (This study revealed evidence of widespread HEV circulation in Thailand. Hepatitis E is not currently a major public health problem in Thailand, but outbreak reports in many countries in Asia, including the neighboring countries of Vietnam and Myanmar, suggest that outbreak prevention and disease awareness of hepatitis E in Thailand should be enhanced (


For transplantation group, BMSCs were injected into the perihemorrhage area in the base ganglia through an intracranial drainage tube 5.5 (3.01–6.89) days after surgery, followed by a second injection into the subarachnoid space through lumbar puncture 4 weeks later. Neurologic impairment and daily activities were assessed with National Institute Stroke Scale (NIHSS), Barthel index, and Rankin scale before transplantation and 6 months and 12 months after transplantation. Our results revealed that, compared with control group, NIHSS score and Rankin scale were both significantly decreased but Barthel index was increased in transplantation group after 6 months. Interestingly, no significant difference was observed between 12 months and 6 months. No transplantation-related adverse effects were investigated during follow-up assessments. Our findings suggest that surgery combined with autologous BMSCs transplantation is safe for treatment of ICH, providing short-term therapeutic benefits.Bone marrow stromal cells (BMSCs) may differentiate into nerve cells under a certain condition; however, the clinical application for treating nervous system disease remains unclear. The aim is to assess the safety profile, feasibility, and effectiveness of surgery combined with autologous BMSCs transplantation for treating ICH. 206 ICH patients who had received surgical procedure were divided into transplantation ( ICH ranks the third leading cause of death, following cardiovascular disease and malignant tumors. Most cases with ICH have various neurological deficits, including aphasia, hemiparalysis, and sphincter abnormalities . Repair Stem cell transplantation and cell engineering have become candidates for treatment of various nerve injuries . CurrentA group of ICH patients were treated with autologous BMSCs transplantation after surgery in the Department of Neurosurgery, Liaocheng People's Hospital of Taishan Medical University, China, based on the previous results . The tren = 431) or refused to participate in this study (n = 109). 215 ICH patients were enrolled in this study. Informed consent was obtained from all participants or their authorized clients.Transplant procedures were approved by the Ethics Committee of Liaocheng People's Hospital . From January 2009 to September 2011, 755 cases with acute ICH were enrolled in the Department of Neurosurgery. 540 cases were excluded for not meeting the criteria (The inclusion criteria used for the study were (1) <80 years old; (2) Glasgow Coma Scale score of 5–12; (3) ICH location on CT scan being limited in the brain basal ganglia; (4) indications for decompression surgery: ICH volume > 20 mL on CT scan and > 10 mm shift of brain from the midline.The exclusion criteria used for the study were (1) ICH caused by factors such as head injuries, anticoagulants, or tumor, excluding hypertension; (2) a history of allergy; (3) mild ICH focal neurological deficits with no indication for decompressive craniotomy or surgical evacuation of hematoma; (4) concurrent chronic illnesses such as hepatic or renal dysfunction; (5) coagulation disorders; and (6) body temperature being more than 37.5 degrees before transplantation.After surgical drainage and decompressive craniotomy, the selected patients or their relatives were further consulted on the potential benefits and risks of this BMSCs treatment, particularly on the uncertainties in its clinical effects and long-term side effects. In the end, full written consent on the cell implantation was obtained from 114 patients. The other 101 patients who did not wish to receive BMSCs treatment participated in this study as the control group.After preoperative examinations, detailed surgery plans were designed for the individuals based on their clinical information and hematoma volume. Indwelling drainage tubes were maintained in the cavities of cases receiving hematoma evacuation via small bone window craniotomies or bone flap craniotomies.Bone marrow aspiration was operated by a hematologist under local anesthesia in a class 100 laminar flow operating room, 5.5 (3.01–6.89) days after surgery. Autologous bone marrow (200 mL) was collected with sodium citrate to prevent clotting and added in stem cell isolating reagent  on a superclean worktable. The cells were centrifuged at 1100 g for 30 minutes. The cells in the interphase were recuperated and washed twice with PBS (300 g for 10 minutes). A stem cell suspension (0.5 mL) was obtained after removing red cells, centrifuging, harvesting, and flushing.The harvested cell suspension in a volume of 0.25 mL was diluted 1 : 20 with saline. An aliquot (0.5%) was removed at 4°C in the neurological laboratory to count nucleated cells and BMSCs, with the rest being used for transplantation. Stem cells were administered through indwelling drainage tubes, which had been drawn to the rim of the hematoma cavity before injection and were removed after injection.α-MEM  (2–7.5 × 106 cells/mL) at 10% of fetal bovine serum  and inoculated (2.5–10 × 105 cells/cm2); medium was changed 2-3 d later to remove nonadherent cells, followed by a subsequent change 3–5 d later. Digestion passage was performed when confluence attained 80%. Microbial levels, cell counts, and survival rates were independently examined by a third party to ensure culture quality. Cells were harvested and counted at passage 3 , CD29-FITC (Cat: 555005), CD44-PE (Cat: 550989), and CD166-PE (Cat: 559263). All antibodies were purchased from Becton Dickinson . Then cells were washed 3 times with phosphate buffered saline (PBS). Cells were measured by FACSCalibur (BD Biosciences).Six months after BMSCs implantation, the patient's swallowing difficulty (Kubota water experiment), muscle strength (MMT), muscle tension (MAS), language ability (Part “V” of GCS), calculation , cognition (MoCA), and responsiveness to painful stimulation (Part “M” of GCS) were separately evaluated by a neurologist who was blinded to patient's treatment according to the international uniform scale.NIHSS score, Barthel index, and Rankin scale were assessed before transplantation and 6 and 12 months after transplantation. Statistical comparisons were performed on results of the transplantation group obtained before and after transplantation and between the transplantation and control groups. The safety of autologous BMSCs transplantation was monitored using cranial CT or MRI, blood routine examination, biochemical indices, and tumor biomarkers  before transplantation and at 6 and 12 months after the transplantation.  t-test or nonparametric Mann-Whitney test, and P < 0.05 was considered as statistical significance.Data were presented as the mean ± SEM or median (interquartile range). SPSS software package  was used for the statistical analysis. The distribution of the samples was determined by Kolmogorov-Smirnov test. The data from experiments were analyzed by Student'sP > 0.05, P > 0.05, Four patients from the implantation group and five from the control did not return to the hospital for schedule follow-up, who were excluded from the final analysis. 110 patients in the implantation group and 96 patients in the control group were included in the final analysis. There was no significant difference in the demographic data including age, sex, neurological findings, and the mean volumes of bleeding between the two groups . There were 12 cases and 3 cases of complete recovery of all symptoms in MSC group and control group, respectively . No significant difference in the scores was observed between 12 months and 6 months  from the transplantation group were identified with low grade fever (<38.5°C) in 24 h after the first transplantation. Two patients (1.81%) also experienced low grade fever after the second transplantation. The fever in all 9 patients subsided in 3 days without specific pharmacological intervention. Except for one patient (0.91%) in the transplantation group with an increase in CA-153, no abnormalities in brain CT or MRIs , routineBone marrow stromal cells (BMSCs) used in our study have significant advantages: (1) ease of obtaining large numbers of cells with bone marrow aspiration; (2) fast amplification in a short period; (3) the probability of these cells crossing the blood brain barrier ; and (4)The safety of the transplantation is our utmost concern. In a study performed by Rice et al. , autolog7 via three generations of culture. As a result, 91.9% (101/110) of transplantation group cases showed improved performance and quality of life 6 months after surgery, characterized by lower NIHSS scores and higher Barthel's scores. Meanwhile, the cases (n = 9) without significant improvement were not worse than before. Comparisons between follow-up results obtained 6 and 12 months following transplantation showed no significant differences, even a moderate decrease in Barthel's scores 12 months after transplantation indicating that the effective window of recovery was not prolonged. We speculated that some changes have occurred with time prolongation, such as neural plasticity, functional reorganization, lesions gliosis, and scar formation. BMSCs might not play an important role in this procession. We suggested that BMSCs should be transplanted within 6 months for the best effect.The curative effect of BMSCs transplantation for nerve injuries occurs via multiple pathways. First, transplanted BMSCs migrate towards the lesion site and fuse into tissues, replace the damaged cells, repair and recreate the neural circuitry, and thus repair neurological functions. Second, the interactions between BMSCs and host nerve tissue may generate cytokines that improve neurological function recovery. Sixty ICH cases that receive autologous BMSC transplantation have improved quality of life in a study conducted by Li et al. . In our  in situ transplantation is impossible, injection into the subarachnoid space through a lumbar puncture provides another convenient, low-damage, and repeatable strategy for transplanting stem cells. All 110 cases in the transplantation group received cells via a lumbar puncture to the subarachnoid space for their second transplantation as a means of eliminating the need to repeatedly indwell a tube.The location of grafts relative to lesions was suggested by Kelly et al. to be critical for the survival of BMSCs transplanted for treatment of neurovascular diseases . Current in situ transplantation and possible injury due to placement of a second indwelling. We did also perform the clinical evaluations 3 weeks after first infusion, such as NIHSS and Barthel's score, and found that there was no significant difference compared with the control group, although the value was higher than that of control group (data not shown). Therefore, we did the second infusion. Since the BMSCs were acquired with a low yield of 105 cells, we amplified the number of cells to 107 via three cycles. Transplantation of these increased cells into the subarachnoid space about one month after surgery is between the acute phase and the formation of glial scar, and this helps to solidify and improve the curative effect.The optimal time for transplanting BMSCs for treatment of ICH remains unclear, although the data from animal and clinical models suggest that BMSCs can survive when administered during a range of pathological stages –20. We sBMSCs transplantation provides a new approach for the treatment of neurological dysfunction resulting from ICH. Our result revealed that this study was a success of large scale clinical trial on BMSC transplantation in ICH patients, which is in line with previous study in the therapy of ICH , 24. How


The etiologies of conjugated hyperbilirubinemia in infancy are diverse.Determine the prevalence rates of the specific etiologies of conjugated hyperbilirubinemia in infancy.EMBASE and Pubmed were searched electronically and the bibliographies of selected studies were search manually. The search was conducted independently by two authors.(1) prospective or retrospective case series or cohort study with 10 or more subjects; (2) consecutive infants who presented with conjugated hyperbilirubinemia; (3) subjects underwent appropriate diagnostic work-up for conjugated hyperbilirubinemia; (4) no specific diagnoses were excluded in the studied cohort.Patient number, age range, country of origin, and categorical and specific etiologies.From 237 studies identified, 17 studies encompassing 1692 infants were selected. Idiopathic neonatal hepatitis (INH) occurred in 26.0 % of cases; the most common specific etiologies were extrahepatic biliary atresia (EHBA) (25.89 %), infection (11.47 %), TPN- associated cholestasis (6.44 %), metabolic disease (4.37 %), alpha-1 anti-trypsin deficiency (4.14 %), and perinatal hypoxia/ischemia (3.66 %). CMV was the most common infection identified (31.51 %) and galactosemia (36.49 %) was the most common metabolic disease identified.Major limitations are: (1) inconsistencies in the diagnostic evaluations among the different studies and (2) variations among the sample populations.INH is the most common diagnosis for conjugated hyperbilirubinemia in infancy while EHBA and infection are the most commonly identified etiologies. The present review is intended to be a guide to the differential diagnosis and evaluation of the infant presenting with conjugated hyperbilirubinemia. In the newborn period and in early infancy, cholestatic jaundice, or conjugated hyperbilirubinemia, results from hepatobiliary dysfunction. The prevalence of this disorder is estimated at 1 out of every 2500 live births; extrahepatic biliary atresia (EHBA) and idiopathic neonatal hepatitis (INH) account for two thirds of cases of infantile cholestatic jaundice [Most studies of infantile cholestatic jaundice focus on EHBA. Although this disorder is rare , timely diagnosis and surgical intervention are required to avoid liver failure and death [Excluding EHBA and INH, a myriad of potential etiologies for infantile cholestatic jaundice have been identified; these can be categorized into obstructive and intrinsic processes. Obstruction can be grossly structural or microscopic in nature . Intrinsic processes affect hepatic cellular function  . Given tThis study involved the use of published, summary data from the world’s literature. There was no contact with primary, patient- identified, source data. Temple University's Human Research Protection Program has declared that evaluations of publically available, secondary databases of de-identified patient data are not subject to its review.This study followed the PRISMA guidelines .a priori. The inclusion criteria for this review were:prospective or retrospective case series or cohort study with 10 or more subjects;consecutive infants who presented with conjugated hyperbilirubinemia defined as elevations of total and direct serum bilirubin concentrations beyond the immediate newborn period;appropriate diagnostic work-up for conjugated hyperbilirubinemia and;no specific diagnoses were excluded in the studied cohort.The study protocol was developed by the authors The main search terms were “conjugated hyperbilirubinemia” or “cholestatic jaundice” and “neonates” or “infants.” The following filters were used: human. No language filter was used; however, when abstract review was required, citations without English abstracts were eliminated. The searches were performed independently by two of the authors and the results were compared.Initial evaluation of each article was performed by author LEG and then reviewed by author MTD. Differences in judgment were resolved first by consensus; ties were adjudicated by the third author (SCA).For each selected study, the following information was recorded: number of patients, age range of patients, and country of origin. The categorical and specific etiologies of conjugated hyperbilirubinemia and the number of patients classified into each group for all selected studies were recorded. The categories of disease included EHBA, INH, total parenteral nutrition (TPN)—associated cholestasis, Alagille’s syndrome, alpha 1-antitrypsin deficiency, cystic fibrosis (CF), infection, hypopituitarism/hypothyroidism, progressive familial intrahepatic cholestasis (PFIC), perinatal hypoxia/ischemia, interlobular bile duct paucity, inspissated bile syndrome, choledochal cysts, hemolysis, metabolic diseases, and others. Within the categories of infection and metabolic diseases, specific etiologies were catalogued based on the data provided in each study. Data collection was reviewed by the second author (MTD).Results from all of the studies were pooled to provide rate estimates of categorical and specific etiologies of cholestasis. Rate estimates for etiologic categories were calculated from the number of subjects used in the entire review. Rate estimates for specific infectious and metabolic etiologies used the total number of subjects reported with specific infectious and metabolic disorders as the denominator.Variability in diagnostic evaluations and the potential for overestimation of INH represent possible sources for bias. The identification of multiple etiologies in individual patients as well as inconsistencies and vagaries in nomenclature were potential sources of error when studies were combined. In some cases, delayed clinical presentation may have affected diagnostic accuracy.The results of the literature search are shown in the Fig. The 17 studies that met the inclusion criteria are presented in Table The etiologies of infantile conjugated hyperbilirubinemia, by study, are shown in Table The etiologies of conjugated hyperbilirubinemia in infancy were defined categorically, by process, and by specific disease entity, where adequate data existed. The categorical etiologies are shown in Table Pseudomonas aeruginosa, Staphylococcus species, Klebsiella species, E. coli, cocksackie B, and parainfluenza type 3 [The specific infectious etiologies associated with infantile conjugated hyperbilirubinemia are shown in Table a type 3 , 17, 20.Metabolic disorders associated with infantile conjugated hyperbilirubinemia are shown in Table Study sizes ranged from 20 to 249 subjects; the largest study accounted for 14.7 % of the total sample reducing the risk of selection bias in the pooled results. The subjects represented 12 countries and five continents; 2 studies drew patients from entire regions and the remaining studies each represented one clinical site. Eight of the centers were referral sites. Nine of the studies were prospective and eight were retrospective. Five studies focused on specific diagnostic techniques to differentiate biliary atresia from neonatal hepatitis.Tiker et. al and Ipek et. al studied subjects admitted to neonatal intensive care units , 17. In Vagaries associated with terminology, e.g., “neonatal hepatitis,” “idiopathic neonatal hepatitis,” and “cholestatic jaundice,” were encountered in multiple studies , 12, 13.Concurrent diagnoses were also a source of bias across studies. Ipek et. al categorized each subject under a single etiology but reported that “the majority of infants (80.4 %) had concomitant clinical disorders that might have contributed to the development of conjugated hyperbilirubinemia.”  AanpreunAlthough Downs syndrome is not a proven cause of conjugated hyperbilirubinemia, it was cited as the etiology in 4 studies , 10, 13.Infantile conjugated hyperbilirubinemia presents with persistent jaundice as part of a clinical constellation that may include other symptoms based on the underlying etiology. The differential diagnosis is broad and requires timely evaluation . The datThe diagnostic evaluation should be guided by symptomatology and may include various imaging studies and serologic, hematologic, and urine investigations for various infections and endocrinopathies, as well as genetic testing for inborn errors of metabolism. While the definitive diagnosis of EHBA requires a percutaneous liver biopsy, ongoing research is investigating less invasive methods of differentiating EHBA from other etiologies of infantile conjugated hyperbilirubinemia .Inconsistency of the diagnostic approach is a major limitation of this review. While there are general guidelines directing the evaluation of an infant with conjugated hyperbilirubinemia, reports published prior to establishment of these guidelines are included in the present review . MoreoveVariability in sample populations is also a potential source of bias. While 12 countries and 5 continents are represented, there is no data from Eastern Europe or South America. Disorders that may be uniquely prevalent in these areas may be underrepresented. Finally, summary data that includes subjects from all over the world may be less relevant to any specific country given the uneven distribution of etiologies between developed and developing countries as well as diseases endemic to the East and West.Finally, the use of the category of INH to include all idiopathic cases of infantile conjugated hyperbilirubinemia is a potential source of bias in this review , 18, 19.The etiologies of infantile conjugated hyperbilirubinemia are numerous. Because of the consequences of untimely correction of EHBA as well as the potential consequences of untreated galactosemia, hypothyroidism and other etiologies associated with this problem, clinical evaluation needs to be prompt, focused, and complete. While specific symptoms may narrow the diagnostic possibilities, a complete history and physical examination, diagnostic imaging and laboratory investigations directed at the more common etiologies is required to make a prompt, definitive diagnosis. This systematic review provides evidence to direct the investigation of an infant with conjugated hyperbilirubinemia.


EPR tooth in vivo dosimetry is an attractive approach for initial triage after unexpected nuclear events. An X-band cylindrical TM010 mode resonant cavity was developed for in vivo tooth dosimetry and used in EPR applications for the first time. The cavity had a trapezoidal measuring aperture at the exact position of the cavity’s cylindrical wall where strong microwave magnetic field H1 concentrated and weak microwave electric field E1 distributed. Theoretical calculations and simulations were used to design and optimize the cavity parameters. The cavity features were evaluated by measuring DPPH sample, intact incisor samples embed in a gum model and the rhesus monkey teeth. The results showed that the cavity worked at designed frequency and had the ability to make EPR spectroscopy in relative high sensitivity. Sufficient modulation amplitude and microwave power could be applied into the aperture. Radiation induced EPR signal could be observed remarkably from 1 Gy irradiated intact incisor within only 30 seconds, which was among the best in scan time and detection limit. The in vivo spectroscopy was also realized by acquiring the radiation induced EPR signal from teeth of rhesus monkey whose teeth was irradiated by dose of 2 Gy. The results suggested that the cavity was sensitive to meet the demand to assess doses of significant level in short time. This cavity provided a very potential option for the development of X-band in vivo dosimetry. Radiation disasters such as Three Mile Island incident, Chernobyl and Fukushima stricken nuclear reactors, vividly illustrate that populations potentially expose to unknown doses of ionizing radiation, especially terrorisms such as the release of dirty bomb probably cause great panic and disruption in public Radiation induces radical materials in hard tissue, especially the radiation induced signal in human teeth would maintain measurable and stable even after quite a long time In order to meet the desirable triage demand in nuclear disaster rescue, scientists are developing EPR spectrometer especially for in vivo dosimetry, quickly and in situ, which will offer an ideal tool for accident rescue and greatly expand the EPR applications 111 mode cylindrical cavity based on the previous work of Ikeya 010 mode cavity which was more suitable for in vivo tooth measurement. TM010 mode cylindrical cavity has been rarely used in common EPR practice. However, the microwave distribution of this cavity has the superiority to be developed into an outside-measurement cavity for special usage of in vivo tooth dosimetry. Compared with TE111 mode cavity, TM010 cavity costs much shorter scan time and could acquired radiation induced signal from irradiated tooth of lower dose. It would be a significant expansion of current EPR cavity resonators and applications.After re-engineering a TE010 mode cavity was designed and calculated according to theoretic functions, then simulated by finite element software to verify and optimize the parameters. This paper introduces the design principle, microwave configuration and the theoretical calculation of the design process, as well as the feature test by measuring DPPH and 18 intact incisor samples. Radiation induced EPR signal could be observed remarkably from 1 Gy irradiated intact incisor within only 30 seconds, which is among the best in scan time and detection limit of in vivo dosimeters. As a supplement experiment for comprehensive verification, the in vivo rhesus monkey measurement experiments demonstrated how the X-band spectrometer worked and proved this scheme had the potential to be developed into feasible dosimeters.This TM010; 3) higher quality factor Q; 4) as small as possible.The resonant mode, frequency and unloaded quality factor Q were related to cavity’s dimension, manufacture process and cavity wall materials. The following conditions were taken into account when designing the cavity’s configuration: 1) the resonant frequency was about 9.6 GHz to match microwave bridge; 2) the resonant mode was TMr and the height be d. First think about the resonant frequency was 9.6 GHz. The relation between r, d and frequency f of ideal cylindrical resonator was referred byLet the ideal right circular cylindrical cavity radius be rε was relative permittivity, rµ was relative permeability, nm’p was the m-th root of n’J, and nJ was the Bessel function of the first kind. n, m and l were the numbers of half-wavelength variations in the standing-wave pattern of cylindrical TMnml mode 010 mode, among all the modes in cylindrical cavity, worked at the lowest cutoff frequency when its dimension fitted certain conditions. This guaranteed the cavity works at designed lowest frequency, and the cavity was smallest in size for convenience of being placed in mouth. Let n = 0, m = 1, l = 0, f was a function of r dimension:TMd about f010 meant that the resonant cavity could be made as narrow or wide (d dimension) accommodating to the magnet gap and teeth measurement. While taking into account that TE111 mode was usually regarded as the dominant mode in a cylindrical cavity, but TM010 mode worked at lower resonant frequency when:The lack of dependence of r and d dimensions were selected to ensure its resonant frequency was 9.6 GHz. d dimension was slightly larger than the width of standard 3 cm wavelength waveguide (about 10.16 mm) for convenient manufacture and assemble. rε and rµ were referred as air filling.As a summary consideration: the So, the equations were given by010f was only controlled by r but had no relation with d, therefore d was designed that the cavity worked at TM010 mode and was easy to manufacture and assemble as well as convenient to imbed into oral cavity. Let d be 12 mm, the solution of Function 3 showed that Matlab . The constraint conditions were referred to 4. It turned out that the theoretical frequency was 9.58 GHz, quality factor Q under ideal conditions was 8000. And it indicated that 1∼3 mm in depth in the cavity was the most efficient depth of magnetic field H1.The cavity microwave distribution and resonant frequency was calculated by HFSS (HFSS is a microwave simulator by ANSYS corporation) using the following parameters: Material, silver; Filling dielectric, air; Mode, Eigen mode; Cavity Radius, 12 mm; Cavity Height, 12 mm; aperture size, 12 mm×3 mm.The design was aided by finite elements simulation calculations. Its working condition was calculated by HFSS using the following parameters: Analysis, parameter sweep analysis; Sweep parameter, coupling size; Sweep range, linear step, from 1 mm×1 mm to 2 mm×8 mm, step was 0.1 mm×0.1 mm; Scan, frequency sweep from 9.5 GHz to 9.7 GHz, step was 0.01 GHz; Results, the scattering-matrix parameter S11 against frequency.The coupling unit was simulated by HFSS using the following parameters: Analysis, sweep analysis; Sweep parameter, aperture width; Sweep range, linear step from 1 mm to 8 mm, step = 1 mm; Results, eigen mode resonant frequency and quality factor Q.The aperture size was simulated by The cavity was manufactured by professional microwave instruments factory using copper material and polished rigorously and plated by silver for lower surface resistance. The wall’s thickness was about 1.5 mm so that it can supply enough structure strength. The resonant frequency and quality factor Q of the cavity were tested using reflection method by hp 8350B scan oscillator and hp 8757D scalar network analyzer.−4 in a sphere area of diameter 10 mm. Microwave bridge was a Bruker-ER041 unit. Signal processor was the SR830 digital phase-locked amplifier of Stanford Research System. This platform offered in vivo detection conditions and made it possible to investigate the cavity performance.As the foundation of research, we designed an in vivo EPR dosimetry platform including clinical magnet which had relative large gap for in vivo use, modulation coils separated with cavity and fixed on magnet poles, alternating current power amplifier for modulation, signal receiver and data process unit and other relevant equipments. The homogeneity of the magnet was approximately 5×10DPPHhydrazyl) sample was used for signal intensity calibration. DPPH powder was inserted in quartz capillary to form a standard sample of 1 mm in length and 1 mm in diameter. The DPPH sample was fixed in the centre of the cavity’s aperture. Typical experiments conditions were: microwave power 1∼200 mW and fixed at 1 mW when modulation amplitude changed to obtain modulation feature, modulation amplitude 0.11 mT and fixed at 0.1 mT when microwave power changed to obtain microwave power feature, scan time 20 s, time constant 0.04 s, scan width 10 mT, centre field 0.34 T, scan magnetic field width 10 mT.60Co radiation source with different doses of 0 Gy, 1 Gy, 2 Gy, 4 Gy, 6 Gy, 8 Gy.All measurements were performed on a homemade in vivo spectrometer platform. The samples were 18 pieces of irradiated human incisors which had no significant dental cavities or metal filling or history of endodontic treatment. All the teeth were collected from departments of stomatology in hospitals. They were removed as part of routine medicine care. These teeth were abandoned by patients and anonymized. The author had no contact with the patients. Teeth samples were separated into 6 groups, each group had 3 teeth of different volume. Incisors of different groups were irradiated by In order to simulate the in vivo measurements, the teeth were immersed in physiology saline. The teeth were wiped dry and fixed in gum model when measured. Only the cusps of the incisors were fixed into the cavity’s aperture, thus only the enamel part of the tooth was detected. The detection length of the teeth was controlled at about 2 mm, and the detected width and thickness of all teeth’s were recorded. Typical ERP measurement conditions were: scan time, 20∼300 s; scan width, 10 mT; microwave frequency, 9.5 GHz; microwave power, 20 mW; time constant, 0.03 s to 0.1 s.BIRM animal welfare ethics committee. The license No. was IACUC of BIRM-2013011.The experiment animal was a rhesus monkey, 2 years old and conformed to the quality control standard of GB14922.1 and GB14922.2. The housing of the animals, design of the experiments, and other conditions, were approved by the animal welfare ethics committee, which conformed to the legal regulations of China in the responsible conduct of animal studies. The full name of the ethics committee was The monkey was housed in conventional monkey house of animal centre. The conditions were: 24 hours negative pressure ventilation, room temperature 18∼24 centigrade, relative humidity 40%∼70%. The monkey house was twice water washed every day. The food was standard monkey feed offered by animal centre and the drink was disinfected cool water. Toys were applied to the monkey for environmental enrichment.60Co radiation source. The monkey’s front teeth were cleaned up with distilled water, and then disinfected by 75% alcohol solution. The monkey and the microwave cavity were fixed at proper position and posture to make sure convenient measurement therefore the monkey could bit into the cavity aperture by its incisor cusp. The monkey was not sacrificed. The detection EPR parameters were: microwave power, 10 mW; scan time, 100 seconds; modulation amplitude, 0.2 mT; time constant, 0.03 s; center field, 338 mT; scan magnetic field width, 10 mT.The monkey was anesthetized with 3% sodium pentobarbital (1 ml/kg). The monkey’s teeth were partially irradiated with dose of 2 Gy by 1 concentrated and weak microwave electric field E1 distributed. The cavity resonator was composed of cylindrical cavity, detection aperture, tune module and a lid assembled by screws. The tune module consisted of coupling hole and a dielectric screw with a metal tip to adjust coupling coefficient. There was a trapezoidal measuring aperture penetrating the metal wall at the exacted position where strong microwave magnetic field H0), and could comfortably and effectively encompass the human head chin. The distance between the magnet poles was 90 mm. The red coils attached to magnet poles provided modulated magnetic field Bm, and was driven by specially designed alternating current amplifier.010 mode had been rarely used in regular EPR spectrometer for its electromagnetic distribution was not appropriate for conventional applications. However it applied an ideal option to make measurements when the sample was out of the cavity. 1 was paralleled to XY plane and most energy was distributed near the cavity cylindrical wall. 1 was paralleled to Z axis, and concentrated within the center area of the cavity. 1 direction was induced by the magnetic field H1 tangential to the cavity walls. The circuit flowed through the centers of the magnetic field loops, and induced charges on the XZ plane near the centers of these loops. The energy density of E1 reached the highest in the center of the cavity but was relatively weak near the cylindrical wall. HFSS in the XY plane in middle cavity region. The simulation results suggested that: (1) the microwave electromagnet configuration was accordant with 010; (3) resonant frequency was 9.58 GHz, and (4) unload quality factor Q in ideal condition was about 6500. These results were quite close to our theoretical calculations of ideal resonant cavity. Besides, the next lower resonant frequency was 14.50 GHz, which leaved a sufficient safe region for single mode demand.TM1 applied into the aperture was perpendicular to the scan magnetic field and modulation magnetic field; (2) the aperture was opened where there was strong H1 and weak E1 field; (3) the aperture’s longer edge was parallel to current flow on the inside cavity wall avoiding too much cutting of surface current flow. The first requirement arose from the nature of the resonance condition. The second requirement was because the amount of microwave energy absorbed by the sample was proportional to H12 (before saturation) and the greater of the H1, the higher of the signal-to-noise ratio could acquire from spectra. And the third requirement minimized microwave energy loss The dot-line rectangle in The resonant cavity was coupled to the waveguide by coupling hole. The detection aperture’s geometry was inevitable for detection sensitivity. Simulation calculations could only roughly offer reference in designs, however the best coupling and aperture was determined by many factors. These parameters were optimized in the following real experiments under different parameters.Some main physical parameters of the cavity were listed in Experiments were carried out by measuring DPPH sample with different microwave power and modulation amplitude. Three characteristics of the cavity were interested: 1) Whether EPR signal could be acquired using this cavity and relevant equipments; 2) Whether sufficient microwave power could apply on sample in the aperture; 3) Whether sufficient modulation magnetic field could apply into the aperture.As the result, the EPR spectrometer acquired strong signal of DPPH samples, certifying this homemade spectrometer worked well with this cavity. The measurements of intact incisors were performed to verify the applicability of in vivo tooth dosimetry using this cavity, and check its detection sensitivity and signal-dose response. The teeth were fixed in gum model to simulate in vivo measurement. The position and depth of the incisors into the aperture were controlled accurately, and then recorded their effective detection sample volumes. EPR spectra were recorded at routine laboratory environment. The in vivo EPR spectrometer with this cavity measured all the 18 irradiated teeth which were radiated with dose of 0–8 Gy  and afteThe rhesus monkey teeth were measured to verify its applicability of tooth dosimetry under the conditions that were most analogous to in vivo usage. The monkey was irradiated with dose of 2 Gy. 010 mode cavity was designed as the detection probe for in vivo tooth dosimetry. Apparently the most important future was whether it could perform in vivo measurement and sensitive enough to detection weak radiation induced signal in irradiated tooth. X-band in vivo dosimetry was usually regarded infeasible for its microwave attenuation caused by water and lower cavity sensitivity since Ikeya’s effort. In order to confirm that, we measured teeth fixed in a gum model, and were immersed in physiological saline before measurement to simulate the moist environment in human oral. Most notably, the detection area in our method is the incisor cusp, which is mostly made of enamel instead of dentine and contains very little water and brings little microwave attenuation. The soft tissue such as the dental bed is far from the detection aperture, the dielectric loss from which could be ignored.This TMThe most fascinate advantage of X-band in vivo dosimetry is the higher detection sensitivity. Its radiation induced signal is much easier to distinguish and analysis. L-band EPR dosimeter, such as the apparatus developed in Dart-Dose CMCR, can detect tooth spectra in less than 300 seconds data acquisition to estimate dose with standard error of less than 1 Gy How scan magnetic field and modulation magnetic field applies is another obstacle for X-band in vivo condition. We improved and redesigned the scheme of X-band in vivo spectrometer, including the cavity, C-shape magnet, modulation amplitude and coils, signal process unit and so on. The modulation magnetic field was excited by coils far away from the cavity, which helped to simplify the cavity structure and improved resonant quality factor Q. During the experiments all the spectra were acquired by this spectrometer and indicated this scheme was quite feasible. We measured a rhesus monkey tooth to initially but comprehensively check our scheme and the feasibility of in vivo spectrometer. Systematic in vivo measurement experiments, such as dosimetry of patients who received radiotherapy or volunteers whose tooth is replaced by an ionized tooth of known dose, will be carried out in the future investigation.The signal-dose relation is a rather complicated procedure. The signal intensity is influenced by many factors such as radiation dose, tooth age, tooth size, tooth angles, detection position, volume alignment, spectrometer parameter and so on. We measured the tooth size and rigorously controlled the effective detection volume to insure the signal intensities of different teeth comparable. More precise detection experiment will be carried out in the future by adding reference sample in the cavity and by some other implements.010 mode cavity has not been reported in EPR study before, and this novel approach also expands EPR resonant cavity family. Our initial work mainly focuses on the development of more sensitive cavity. Further detailed design of cavity will focus on aperture geometry considering incisors match, structure simplify, cavity filling factor, and reference signal label. The future research of spectrometer will also pay more attention on spectrometer improvement, human in vivo measurement experiment and signal-dose effect.TM


In previous years, research has focused on the situation of psychiatric patients' minor children. The aims of this qualitative study were to describe the experience of adult children of depressed and bipolar patients, including positive and negative factors as well as coping mechanisms, and to investigate possible predictors of burden in order to identify children in need of professional support.Freiburger Fragebogen zur Krankheitsverarbeitung, FKV). Regression analysis indicated the most relevant predictors of burden.A total of 30 adult children were interviewed using a semi-structured interview. In addition, all children completed the Freiburg Questionnaire of Coping with Disease (All (100%) of the children reported emotional burden due to the illness of their parent, 90% suffered from impaired family life, and 77% experienced burden due to the parent's symptoms. Reward (positive experience) was reported regarding the intensification of the parent-child relationship. Linear regression analysis shows predictors for highly burdened children as well as for children who are more prone to maladaptive ways of coping. Higher burden was significantly associated with the child's age, severity of illness of the parent, and specific diagnosis.Although some positive aspects of parental affective disorder exist, this study underlines that children primarily suffer from their parent's disorder and that this burden does not stop in adulthood. Providing professional support to adult as well as to minor children of affected individuals should become standard of care in clinical settings. Typically, the term ‘a sample of caregivers’ refers to parents and partners but rarely to children. In studies exploring caregiver burden, children of psychiatric patients tend to comprise a small part of a heterogeneous subject group  and of adult offspring of parents with depressive disorder (ODD).for the children (ODD vs. OBD). The tested characteristics are gender, age, level of graduation, employment, marital status, common household with the patient, number of persons in the household, monthly take-home amount, hours of contact with the patient per week, distance between place of residence and clinic, legal guardianship of the patient, and taking part of a psychoeducational caregiver group. The only significant difference in the group comparisons in the characteristics for the patients (ODD vs. OBD) was in the psychosocial functioning (GAF). The OBD group had a significantly lower psychosocial functioning at discharge than the ODD . The tested characteristics are gender, age, level of graduation, employment, duration of illness, psychotherapy before admission, history of suicide attempts, clinical global impression (CGI part 1) at admission, global assessment of functioning at admission and discharge (CGI), legal guardianship, mentally ill, first-degree family members, comprehension of caregivers in patient's treatment, and situation of living of the patient after discharge.Patients with either bipolar disorder or depressive disorder (ICD-10 F31.x and F32.x/F33.x) and who had at least one adult child were recruited consecutively 6 months along to participate in the study during their psychiatric hospitalization. Patients and adult children were informed about the study and were offered to participate voluntarily. The study protocol was approved by the ethics committee of the Medical Faculty of the University of Regensburg, and all participants gave their written informed consent to participate. One hundred twenty-four patients were contacted: 6 did not want any contact with our interviewer, 40 stated not to have children, 15 patients only had minor children, and 63 had adult children who could be included in our study. Of these 63 patients who were asked for permission to contact their adult child/children, 33 (52.4%) agreed and gave their written informed consent. Of the 33 children contacted, 30 (90.9%) agreed to participate in the study and 3 (9.1%) did not. Those 30 children came from 30 separate patients. The socio-cultural-economic background of the participants (patients as well as interviewed children) is comparable. There are no significant group comparisons in the characteristics Clinical records of the patients were complemented with socio-demographic variables assessed through a small questionnaire answered by all patients.The assessment of the adult children involved a self-generated interview using a semi-structured schedule assessing both objective and subjective burden of care. Furthermore, the schedule included one question regarding reward: ‘Did you experience any positive changes in your life due to your parent's illness?’ All interviews were done by one rater (psychologist), trained by experienced psychologists in different workshops in psychological interviewing. The interviews were audio-taped and transcribed. In addition, the Freiburg Questionnaire of Coping with Disease  and the offspring of patients with depressive disorder (ODD), R2 were carried out to identify the variables which explained best the variance of burden and coping strategies. All clinical variables (inclusive diagnosis) regardless of the group differences were included as independent variables. The three dependent variables that were used were the sum score of reported burden, and the coping strategies ‘Depressive coping’ and ‘Trivialization and wishful thinking’, which were seen as maladaptive coping strategies.Linear regression analyses built by backward elimination and model refinement on the basis of the adjusted α ≤ .05. Statistical analysis was performed with the Statistical Package for Social Sciences (SPSS) software, version 16.0.Statistical significance was set at The patient sample consisted of 19 female (63.3%) and 11 male (36.7%) patients of whom 9 (30%) were 49 years or younger, 10 (33.3%) were between 50 and 59 years old, and 11 (36.7%) were older than 60 years. Fifteen (15) patients had bipolar disorder, and 15 had depressive disorder. The children sample consisted of 20 (66.7%) female and 10 male (33.3%) persons between 18 and 46 years old. Demographics for patients and their children are reported in Table Duration of the interviews was 13 to 71 min with an average of 28.6 min.The children's statements were summarized into 38 global statements and assigned to eight categories of burden: emotional burden (reported by 100%), burden due to impaired family life (90%), burden due to the patient's symptoms (76.7%), burden due to dissatisfaction with the patient's therapy/professional staff (73.3%), burden due to impaired free-time activities (63.3%), burden due to impaired functioning in school/job (46.7%), burden due to own health problems (43.3%), and burden due to problems in the child's relationship/own family (30.0%). Categories and statements are more precisely listed in Table t (N = 30) = −1.55; p = .97]. However, OBD reported significantly more burden due to impaired functioning in school/job , whereas ODD reported significantly more burden due to dissatisfaction with the patient's therapy/professional staff .There was no significant difference between the sum score of the OBD and the ODD , although OBD reported significantly more reward due to feeling closer to the healthy parent .There was no significant difference between the sum score of the OBB and the ODD [p = .01) and also more ‘Distraction and building self-esteem’ (p = .05) than ODD.OBD and ODD of this sample used a wide range of coping strategies, none of which was predominant. Nevertheless, OBD used significantly more ‘Depressive coping’ , adult child being of younger age , adult child reporting more burdensome aspects , adult child being in a cohabitating relationship , and parent being diagnosed with bipolar disorder . The variables explained 59% of the total variance .Five factors were found to predict ‘Depressive coping’: ill parent being single , parent being single , parent having attempted suicide , parent being diagnosed with bipolar disorder , adult child being 10 years or younger at the onset of the parental illness , and adult child being the parent's legal custodian . The variables explained 71% of the total variance .Six factors were found to predict ‘Trivialization and wishful thinking’ best: parent  not being under legal tutelage Child was 10 years or younger at the onset of parental disorderChild has a cohabitating relationshipChild is the patient's legal custodianChild reports many burdensome aspectsParent is diagnosed with bipolar disorderParent has a severe disorderParent is singleParent is not under legal tutelageParent has attempted suicideHigh burden due to a caring for a family member with a psychiatric disorder leads to a higher risk for developing affective disorders in caregivers  or part time (20%); 10% were housewives. None of them receive a pension, and the income level was relatively high. Furthermore, subjects were recruited primarily from a hospital setting and included only children who were willing to participate. These findings are, thus, limited in their applicability to other populations such as children of parents with less severe or less chronic conditions.We have increased our knowledge about the clinical importance of supporting children of mentally ill parents. Introducing psychoeducational interventions and, as a result, reducing burden and promoting problem-oriented coping will not only diminish the children's own risk of developing psychiatric disorders but also improve the clinical outcome among patients with affective disorders (Perlick et al. Although some positive aspects of parental affective disorder exist, this study underlines that children primarily suffer from their parent's disorder in important areas of their life and that this burden does not stop in adulthood. Providing professional support to adult as well as to minor children of affected individuals should become standard of care in hospital settings.


To assess the capacity of methylxanthines  to inhibit uric acid crystallization, and to evaluate their potential application in the treatment of uric acid nephrolithiasis.The ability of methylxathines to inhibit uric acid nucleation was assayed turbidimetrically. Crystal morphology and its modification due to the effect of theobromine were evaluated by scanning electron microscopy (SEM). The ability of theobromine to inhibit uric acid crystal growth on calculi fragments resulting from extracorporeal shock wave lithotripsy (ESWL) was evaluated using a flow system.The turbidimetric assay showed that among the studied methylxanthines, theobromine could markedly inhibit uric acid nucleation. SEM images showed that the presence of theobromine resulted in thinner uric acid crystals. Furthermore, in a flow system theobromine blocked the regrowth of post-ESWL uric acid calculi fragments.Theobromine, a natural dimethylxanthine present in high amounts in cocoa, acts as an inhibitor of nucleation and crystal growth of uric acid. Therefore, theobromine may be clinically useful in the treatment of uric acid nephrolithiasis. Renal lithiasis is a highly prevalent condition, currently affecting about 10% of the worldwide population Uric acid is the final product of purine catabolism in humans. In most other mammals, such as rats and dogs, uric acid is further degraded to allantoin by the enzyme uricase Uric acid nephrolithiasis accounts for 7–10% of kidney stones Theobromine is a dimethylxanthine present in high amounts in chocolate and cocoa Studies in healthy volunteers showed that 50% of administered theobromine is recovered in urine after 8–12 h, and 100% is recovered after three days, suggesting that this compound is completely or almost completely absorbed The aim of the present work is to study the inhibitory effect of theobromine on uric acid crystallization in synthetic urine, using different in vitro models. Theobromine concentrations used in the present study were selected according to its normal levels in urine after consumption of theobromine.Uric acid, theobromine, theophylline, caffeine and paraxanthine were purchased from Sigma-Aldrich . Synthetic urine components were obtained from Panreac . Chemicals of analytical reagent-grade purity were dissolved in ultra-pure deionized water from a Milli-Q system and filtered through 0.45 µm pore filters before use. Uric acid stock solution was prepared daily by dissolving 1 g uric acid in 0.5 L of water with 1 M NaOH addition. Synthetic urine was prepared by mixing equal volumes of A and B solutions Table 1,Uric acid crystal formation in synthetic urine and the effects of potential crystallization inhibitors were assessed using a kinetic turbidimetric system, consisting of a photometer (Metrohm 662) equipped with a fiber-optic light-guide measuring cell with an attached light path 2×10 mm reflector and monochromatic light (550 nm). Crystallization was assessed at constant temperature (37°C) with magnetic stirring.Synthetic urine solutions were adjusted to pH 5.40, and 2 g/L uric acid solution was adjusted to pH 10.70. Synthetic urine (160 mL) was added to a crystallization flask, followed by 40 mL uric acid solution. When the resulting solution reached a temperature of 37°C, 1.4–1.6 mL HCl was added to achieve the desired supersaturation of uric acid (pH 4.40–4.70), and the timer was switched on. These pH values were selected in order to have a short crystallization time (6–10 minutes without inhibitor). The pH of the final solution was measured at the beginning of each experiment. The absorbance of the solution (550 nm) was measured every two minutes, until the end of the kinetic assay.Caffeine, paraxanthine and theophylline were assayed at 40 mg/L, and theobromine at concentrations of 10, 20 and 40 mg/L, each at three different pHs . Each experiment was repeated twice in order to assess its reproducibility. In some cases, the crystals formed during the turbidimetric assay were filtered through a 0.45 µm pore filter, dried in a desiccator, and examined by scanning electron microscopy.Fifteen post extracorporeal shock wave lithotripsy (ESWL) fragments of uric acid calculi, of similar weight and morphology, were obtained from a single patient. This study was a retrospective evaluation of a clinical patient sample. The volunteer had provided written informed consent for its clinical samples to be used in scientific studies. This study was approved by the “Comitè d'Ètica de la Recerca (CER)” of the University of the Balearic Islands.The fragments were not pre-treated and were placed in a hermetic flow chamber. This chamber was placed in a larger, temperature-controlled (37°C) chamber, and incubated for 48 h. The experimental flow device used Figure 2The calculi were dried in a desiccator before and after each experiment until they reached constant weight, as determined using a precision balance, and fragment growth was evaluated by measuring the difference in weight. Mean growth rates were determined, and fragment growth was standardized by calculating the relative mass increase, thus avoiding the effects of surface area on growth rate.The morphology of the growing calculi was assessed by scanning electron microscopy.The results presented here indicate that, in vitro, theobromine can markedly inhibit uric acid crystallization. Turbidimetric experiments showed that theobromine can inhibit uric acid nucleation, since the delay in induction time was dependent on theobromine concentration. In contrast, caffeine, theophylline and paraxanthine had no effects. This was surprising, inasmuch as their structures are very similar. However, crystallization inhibitors are extremely selective, such that slight modifications in chemical structure can render inhibitors ineffective.Theobromine concentrations used in this study were similar to those observed in urine after consumption of theobromine The theobromine-induced delay in induction time was higher at higher pH, due to the decreased supersaturation of uric acid at increased pH. Thus, the effect of theobromine at pHs between 5 and 5.5  will be even more important, since the supersaturation of uric acid decreases when urinary pH increases. The increased effect of theobromine at higher pH suggests that urinary alkalinization combined with theobromine may be effective clinically.Theobromine also altered the morphology of uric acid crystals figure 8Using post-ESWL uric acid fragments, we found that theobromine also inhibits the regrowth of these calculi in a concentration dependent manner. Furthermore, the crystals formed on the surface of the uric acid were smaller in the presence than in the absence of theobromine. Thus, theobromine may be useful clinically in preventing the regrowth of uric acid calculi fragments after ESWL.The experiments described in this paper were all performed in synthetic urine. The morphology of the crystals showed in this paper may change in real urine, due to the presence of other substances in urine, such as proteins.In conclusion, theobromine acts as a uric acid crystallization inhibitor, with high clinical potential in the treatment and prevention of uric acid nephrolithiasis. Clinical trials are necessary to assess its inhibitory capacity in vivo.


HFE) gene are associated with excessive iron absorption from the diet, and pro-oxidant effects of iron accumulation are thought to be a risk factor for several types of cancer. Methods: The C282Y (rs1800562) and H63D (rs1799945) polymorphisms were genotyped in 301 oral cancer cases and 437 controls and analyzed in relation to oral cancer risk, and serum iron biomarker levels from a subset of 130 subjects. Results: Individuals with the C282Y allele had lower total iron binding capacity (TIBC)  and higher percent transferrin saturation  than wild type individuals. Iron and ferritin levels approached significantly higher levels for the C282Y allele . Conclusions: Iron biomarker levels were elevated by the C282Y allele, but neither (rs1800562) nor (rs1799945) was associated with oral cancer risk in blacks and whites.Background: Polymorphisms in the hemochromatosis ( HFE gene produces a protein located on the surface of liver and intestinal cells that acts to detect iron levels. C282Y and H63D are the two most common missense polymorphisms in the HFE gene, and they both result in increased cellular iron uptake. The C282Y polymorphism prevents the protein from reaching the cell surface, resulting in an inability to accurately sense iron levels in the body [HFE gene are most common in individuals with northern European ancestry, but are also present in Blacks and Hispanics [The the body ,2,3. Indthe body  and sevethe body ,6. H63D the body , and H63the body ,10. Polyispanics .HFE) gene, which results in excess dietary iron absorption and deposition in various organs throughout the body. Heterozygotes have been reported to have higher mean transferrin saturation and ferritin levels than normal subjects, but rarely develop HH. The more prevalent H63D mutation (rs1799945) rarely causes HH. Hereditary hemochromatosis (HH) is one of the most common genetic disorders in persons of northern European descent that causes excessive iron absorption and storage. The primary cause of HH is the homozygous C282Y mutation (rs1800562) in the hemochromatosis , iron levels, ferritin levels, total iron binding capacity (TIBC), and transferrin saturation were stratified by HFE genotype status in order to determine the degree of iron accumulation in patients with HFE polymorphisms.To determine whether there is an increased risk of oral cancer in individuals with n = 437) were individuals without any prior diagnosis of cancer or present respiratory ailment. The controls consisted of friends, spouses, and spousal family members of cancer patients, or subjects receiving outpatient care for non-malignant related conditions. About 95% of eligible controls consented to participate in the study. Controls were frequency-matched to cases by age (±5 years), sex, race, month of diagnosis of the case subject, and institution. Two controls were sought for each case, although for most cases only one control was identified that met the criteria. Cases were patients diagnosed with primary squamous cell carcinoma of the oral cavity from 1994 to 1996 from three institutions : All stui.e., 20 cigarettes = 4 cigars = 5 pipes = 1 pack [Demographic information as well as information on tobacco use was collected for all cases and controls by administering a structured questionnaire by subject interview. Smoking levels were calculated as the sum of cigarettes, pipes, and cigars smoked according to the equivalents adopted previously, = 1 pack , with ouBuccal cell samples, isolated from cases at a follow-up examination, or normal oral tissue adjacent to the excised tumor isolated at the time of surgery from incident cases, were used for the analysis of polymorphic genotype frequencies in cases, whereas buccal cells were collected for the analysis in matched controls. Buccal cell samples were prepared as previously described . Normal Fasting venous blood samples were obtained from 130 consecutively enrolled subjects at Memorial Sloan-Kettering Cancer Center. Pre-treatment samples were obtained from cases and controls from an antecubital vein into two 10 mL Trace Metal Vacutainer tubes containing heparin as the anticoagulant  using a sterile 19-gauge needle. Blood was centrifuged to separate plasma, erythrocytes, and buffy coat. Erythrocytes were washed in saline three times. Plasma and erythrocyte samples were aliquoted for each assay, and all samples were immediately frozen and stored at −70 °C. HFE C282Y and H63D were genotyped using pre-designed Taqman 5’-exonuclease genotyping assays, according to manufacturer’s instructions. SDS 2.2.2 software was used for automated determination of genotypes . Briefly, allele-specific probes for rs1800562 and rs1799945 were labeled with the fluorescent dyes VIC and FAM. During extension, the 5′-exonuclease activity of the Taq polymerase separates the fluorophore from the non-fluorescent quencher. A post-amplification allelic discrimination run on the ABI 7900HT was used to determine genotype based on the relative amount of fluorescence of VIC and FAM. PCR reactions were carried out in a total reaction volume of 5 µL in 384-well plates using the ABI 7900 HT Sequence Detection System, and the thermal cycling conditions were 50 °C for 2 min, 95 °C for 10 min, and then 40 cycles at 95 °C for 15 s and at 60 °C for 90 s. Individuals involved in genotyping were blind to patient status. Quality control methods for PCR were previously described [escribed . Serum iron was measured spectrophotometrically as previously described ,40. Tota2 goodness of fit tests. Odds ratios (OR) and 95% confidence intervals (CI) of association between cancer and SNP genotype were estimated using Fisher’s exact test. Potential confounding of the association between genotype and cancer risk by known risk factors was explored using Spearman rank correlation analyses and multivariate logistic regression models. Iron biomarker levels were compared between wild type individuals and individuals with the H63D and C282Y polymorphisms using an unpaired t-test.All statistical analyses were performed using SAS statistical software . Both the C282Y and H63D variants were found to be in Hardy-Weinberg equilibrium using χp = 0.22). In Blacks, the mean age of cases was 58 ± 10 y and the mean age of controls was 58 ± 13 y (p = 0.95). The mean pack-years of smoking for White and Black oral cancer cases  was significantly (p < 0.01) higher than the mean pack-years observed for White and Black controls . A significantly higher percentage (p < 0.01) of White cases were heavy drinkers (36.0%) compared to White controls (11.7%). Similarly, heavy alcohol consumption was significantly more prevalent (p < 0.01) in Black cases (49.5%) compared to Black controls (15.8%).The basic demographic profile of the oral cancer case and control subjects is shown in HFE SNPs were consistent with Hardy-Weinberg equilibrium. Among White controls, the minor allele frequencies (MAFs) of the HFE SNPs were 0.12  and 0.03 , which is similar to that in HapMap for individuals with European ancestry . The MAF in the Black controls was 0.03 for rs1799945, which was also similar to HapMap African ancestry from the Southwestern USA (0.02), and 0.02 for rs1800562; this SNP has not been previously reported for Blacks in HapMap.Both versus subjects with at least one variant allele. No association was found between the H63D (rs1799945) and C282Y (rs1800562) SNPs and oral cancer risk with  or C282Y (1 case and 1 control) polymorphisms was low, risk analysis was performed by examining homozygous wild-type subjects isk with  or withoHFE polymorphisms and cancers risk.Compound heterozygosity for the two SNPs was explored, and again no association with oral cancer risk was found for any combination of C282Y and H63D genotypes (results not shown). Because HPV infection is a major risk factor for cancers of the oropharynx, the data was stratified by tumor site excluding cases that might be related to HPV infection (results not shown). With the oropharyngeal cases excluded, no association was observed between either of the HFE C282Y allele exhibited significantly lower total iron binding capacity (p = 0.007) and a higher percent transferrin saturation (p = 0.023) than wild type individuals. Iron and ferritin levels approached significantly higher levels for subjects with at least one HFE282Y allele . For the H63D SNP, iron levels were not significantly different between wild type individuals and individuals with one or more copies of the polymorphic allele. Levels of TIBC, ferritin, and transferrin did not differ significantly by H63D genotype.The effect of H63D and C282Y on iron biomarker levels in 130 primarily white subjects is shown in et al. found significant associations between SNPs in oxidative stress-related genes and cancers of the oral cavity [HFE gene can lead to excessive absorption and an imbalance in iron homeostasis.The annual incidence of oral and pharyngeal cancer is approximately 75,000 cases in the United States , with Bll cavity , but litl cavity . Hepcidil cavity . Hepcidil cavity . HFE cauHFE and oral cancer risk. A study that followed 36 patients with idiopathic hereditary hemochromatosis and documented extrahepatic cancers found that one out of 36 patients (~2.7%) developed oral cancer, which is much higher than the oral cancer prevalence in the general population (~0.1%) [HFE polymorphisms are not a risk factor for oral cancer in Whites and Blacks. The lack of a significant association seen for the C282Y allele, and especially in Blacks may be due to low statistical power, since the prevalence of this SNP was low. Because clinical penetrance of HFE polymorphisms is known to be higher in males [HFE C282Y and H63D genetic variants. These findings are consistent with the fact that genetic variants that cause imbalances in iron homeostasis are independent of other risk factors , so the findings should not be biased based on whether OPC was smoking-related or HPV-related.There is little data on  (~0.1%) . This rein males ,57, casein males . In thisHFE genotypes and oral cancer cases may be because of the way excess iron is distributed in different tissues throughout the body. Although absorbed iron is distributed to all tissues of the body for vital cellular processes, excess iron is deposited differentially in the following tissues (decreasing order of severity): liver, pancreas, myocardium, pituitary gland, adrenal gland, thyroid and parathyroid glands, joints, and skin [HFE polymorphisms and oral cancer may indicate that there is not enough iron deposition in the relevant tissues to cause the cellular injuries that lead to neoplasia. Therefore, the functional characteristics of transferrin receptors and how they are regulated may lend important insights into understanding which types of cancers can result from the pro-oxidant effects of iron accumulation.One explanation for the lack of association between and skin . Iron isand skin . The lacHFE gene polymorphisms and clinically elevated iron levels should be conducted in order to better estimate how HH and iron levels may affect oral cancer risk. Because iron accumulation has only been reported in C282Y and H63D homozygotes and compound heterozygotes, larger case-control studies may be necessary to detect an association between these rarer genotypes and cancers of the oral cavity. In addition, because there are other genes that affect iron levels, it may be necessary to study the interaction of polymorphisms in multiple genes involved in iron regulation. A study by Wu et al. found this to be true for genes involved in oxidative stress; no single polymorphism affected oral cancer risk, but combinations of polymorphisms in the manganese superoxide dismutase, myeloperoxidase, catalase, and glutathione peroxidase 1 genes were associated with significantly increased risk of oral cancer [Another explanation for the lack of association with oral cancer risk may be due to the low clinical penetrance of the C282Y and H63D polymorphisms ,57. Evenl cancer .HFE polymorphisms accounted for <5% of the overall variance that was observed [HFE polymorphisms, which agrees well with the findings in the current study that HFE polymorphisms were not associated with the significantly higher ferritin levels observed in oral cancer cases. It is well known that ferritin levels can increase in response to many factors including chronic disease [In previous studies , signifiobserved . In the  disease , even wh disease . Taken tHFE) gene affects iron metabolism. HFE polymorphisms were unrelated to head and neck cancer risk. These studies demonstrate that a single nucleotide polymorphism in the hemochromatosis (