Effects of Lujingyiqishengxue Pill on Iron Metabolism in Rats with Iron Deficiency Anemia

[Objectives]This study was conducted to observe the effect of Lujingyiqishengxue Pill on iron metabolism in rats with iron deficiency anemia. [Methods] The iron-deficiency anemia rat model was established by feeding low-iron diet. Meanwhile, the rats were given oral gavage of ferrous succinate(0.036 g/kg, positive drug group) and Lujingyiqishengxue Pill(4.4, 2.2, 1.1 g/kg, high, middle and low dose groups), once daily for 42 consecutive days. The body weight of the rats was observed every week, and the peripheral blood[red blood cells(RBC), hemoglobin(HGB), and hematocrit(HCT)]and the iron contents in tissues(the liver, spleen, small intestine, kidney) of the rats were detected after modeling;and serum iron(SI), serum total iron binding capacity(TIBC), transferrin saturation(TSAT), serum ferritin(SF) and serum transferrin receptor 1(TFR1) and other iron metabolism indexes were determined. [Results] Compared with the model group, the high-dose Lujingyiqishengxue Pill significantly reversed the peripheral blood(HGB, HCT) and iron contents of various tissues(the liver, spleen, small intestine, kidney) in rats(P<0.01), and significantly increased SI, TSAT, SF(P<0.01), while the contents of TIBC and TFR1 were significantly decreased(P<0.01). [Conclusions] Lujingyiqishengxue Pill can significantly improve anemia and regulate iron metabolism in rats with iron-deficiency anemia, which provides a pharmacological reference for the clinical application of Lujingyiqishengxue Pill.

Effect of Weikangfu Granule (胃康复冲剂) on the Physiopathologic Figure of Precancerosis of Gastric Mucosain Patients of Chronic Gastritis with Pi-Deficiency Syndrome

To study the physiopathologic basis of Weikangfu Granule （胃康复冲剂, WKFG） in treating precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome （CGPDS）. Methods： One hundred and fifteen patients of CG-PDS who suffered from intestinal metaplasia （IM） and atypical hyperplasia （ATHP） of gastric mucosa, were divided into two groups. The treated group （n=61） was treated by WKFG with its ingredients modified according to the syndrome type of patients. The control group（n=54） was treated with Weishu granule （胃舒冲剂）. The histopathological and subcellular ultrastructural changes were detected by optical microscope, screening electronic microscope, transmission electronic microscope and histochemical staining; the nuclear and mitochondrial ultrastructure of gastric mucosa were analyzed with energy dispersion X-ray analyser and image analysis system. And the changes of cAMP, lipid peroxide （LPO）, superoxide dismutase （SOD） before and after treatment in the treated group were measured and compared with those of the health control group consisting of 15 volunteers. Results： The symptomatic and pathological therapeutic effect in the treated group were significantly superior to those in the control group （P〈0.05）. The contents of Zn, Cu, cAMP, SOD and 3H-TdR LCT in gastric mucosa of the treated group before treatment were all lower than those of the healthy control group, yet all these indexes markedly increased after treatment, while serum LPO level, which increased before treatment was lowered after treatment. All the changes showed statistical significance （P〈0.05 or P〈0.01）. Conclusion： WKFG can reverse IM and ATHP in patients of CG-PDS, and the effect may be realized by way of increasing the level of Zn, Cu, cAMP and SOD in gastric mucosa, promoting cell differentiation, enhancing cellular immunity and reducing oxygen free radicals and lipid peroxidation.

Effects of Different Processed Products of Radix Codonopsis on Intestinal Flora of Rats with Spleen Deficiency

[Objectives]This study aimed to observe the effects of different processed products of Radix Codonopsis on intestinal flora in rats with spleen deficiency.[Methods]Rat models with spleen deficiency were established by bitter-cold purgation method with Radix et Rhizoma Rhei.Normal group,model group,Radix Codonopsis group,fried Radix Codonopsis group,rice-fried Radix Codonopsis group,honey-fried Radix Codonopsis group and bran-fired Radix Codonopsis group were designed.After subjecting to corresponding treatments,the changes in the quantity of intestinal microorganisms of the rats were detected.[Results]There was no significant change in the quantity of intestinal microorganisms of the rats in the normal group and model group.The rats in the rice-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased,and the quantity of Escherichia coli and Staphylococci reduced.The rats in the Radix Codonopsis group,fried Radix Codonopsis groups,honey-fried Radix Codonopsis group and bran-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased(the increases were smaller than those in the rice-fried Radix Codonopsis group),and the abundance of E.coli and Staphylococci reduced,close to normal levels.[Conclusions]Different processed products of Radix Codonopsis have obvious regulation effect on intestinal flora of rats with spleen deficiency,and the regulation effect of rice-fried Radix Codonopsis on rats with spleen deficiency is better than that of Radix Codonopsis,fried Radix Codonopsis,honey-fried Radix Codonopsis and bran-fried Radix Codonopsis.

Ultrastructure and histochemistry of rat myocardial capillary endothelial cells in response to diabetes and hypertension

Insufficient growth and rarefaction of capillaries, followed by endothelial dysfunction may represent one of the mostcritical mechanisms involved in heart damage. In this study we examined histochemical and ultrastructural changes inmyocardial capillary endothelium in two models of heart failure streptozotocin-induced diabetes mellitus (STZ) and NO-deficient hypertension in male Wistar rats. Diabetes was induced by a single i.v. dose of STZ (45 mg/kg) and chronic 9-week stage was analysed. To induce NO-deficient hypertension, animals were treated with inhibitor of NO synthase L-nitroarginine methylester (L-NAME) (40 mg/kg) for 4 weeks. Left ventricular tissue was processed for enzyme cata-lytic histochemistry of capillary alkaline phosphatase (AlPh), dipeptidyl peptidase IV (DPP IV), and endothelial NOsynthase/NADPH-diaphorase (NOS) and for ultrastructural analysis. In diabetic and hypertensive rats, lower/absentAlPh and DPP IV activities were found in focal micro-areas. NOS activity was significantly reduced and persisted onlylocally. Quantitative evaluation demonstrated reduction of reaction product intensity of AlPh, DPP and NOS by 49.50%,74.36%, 20.05% in diabetic and 62.93%, 82.71%, 37.65% in hypertensive rats. Subcellular alterations of endothelialcells were found in heart of both groups suggesting injury of capillary function as well as compensatory processes.Endothelial injury was more significant in diabetic animals, in contrast the adaptation was more evident in hypertensiveones. Concluding: both STZ-induced diabetes- and NO-deficient hypertension-related cardiomyopathy were accompa-nied by similar features of structural remodelling of cardiac capillary network manifested as angiogenesis and angiopathy.The latter was however, predominant and may accelerate disappearance of capillary endothelium contributing to myo-cardial dysfunction.

Effects of zinc deficiency on the brain development in rats

The zinc effects on the learning and memory ability were investigated in the zinc-deficient(ZD)and pair-fed(PF) rats and it was found that(1 ) The rate of body weight gaining and the feed-efficiency ratio were significantly lower in ZD group than in PF gr

A Rat Model of Autologous Oral Mucosal Epithelial Transplantation for Corneal Limbal Stem Cell Failure

Purpose: To establish an animal model of autologous oral mucosa grafting for limbal stem cell deficiency.Methods: The study was carried from August to October2012. Fourteen SD rats were randomly and evenly allocated to study group A and control group B. Limbal stem cell deficiency was established by alkali burn in the right eye of each rat in both groups. Rats in group A received autologous oral mucosa strip transplantation following the chemical burn. Rats in group B did not receive surgery after the chemical burn.Topical antibiotics and dexamethasone were used in all rats.Corneal clarity, corneal fluorescein staining, oral mucosal graft survival, and complications at postoperative days 1,3,7,14 were observed.Results: The oral mucosa strip graft was detached in one rat in group A. Reepithelialization was observed starting from the graft position and was completed within 14 days in the remaining 6 eyes in group A. However, persistent corneal epithelium defect was observed in all eyes in group B, among which corneal melting and perforation was observed in 2 eyes and corneal opacification with neovascularization was observed in the remaining 5 eyes.Conclusion: Autologous oral mucosa strip grafting for limbal stem cell deficiency can be achieved by a rat model following chemical burn. The fate of the transplanted oral mucosal epithelial cells warrants further study.

Studies of the effects of dietary zinc on the immune organs and cellular immunity in the rat

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Regulative Actions of the Chinese Drugs for Tonifying the Kidney on Gene Expression of the Hypothalamic GnRH,Pituitary FSH,LH and Osteoblastic BGP

It is found that the drugs for nourishing yin to reduce pathogenic fire can significantly down-regulate, and the drags for tonifying the kidney to replenish essence can up-regulate mRNA expression of the hypothalamic GnRH, pituitary FSH, LH and osteoblastic BGP, indicating that the Chinese drugs for tonifying the kidney can regulate gene expression of the hypothalamic GnRH, pituitary FSH, LH, and osteoblastic BGP, which is possibly one of the main mechanisms of the Chinese drug for tonifying the kidney, regulating ephebic development process and improving skeletal development in sexual precocity children.

Novel translational rat models of dopamine transporter deficiency

Dopamine （DA） is one of the brain＇s fundamental neurotransmitters. Despite the fact that the dopaminergic synapses constitute less than 1% of all brain synapses, DA is implicated in a number of critical physiological functions and in the pathogenesis of important psychiatric diseases such as schizophrenia, attention-deficit/hyperactivity disorder （ADHD）, Parkinson＇s disease （PD） and others.

MECHANISM OF PROMOTING EFFECTS OF RIBOFLAVIN DEFICIENCY ON CARCINOGENESIS OF NITROSAMINES (EFFECTS ON RAT LIVER GLUTATHIONE CONTENT)

The effects of riboflavin deficiency and simultaneously nitrosodimethylamine (NDMA) given by gastric intubation on the hepatic glutathione (GSH) content were examined in rats. On different days of the experiment, hepatic GSH content of the riboflavin deficient rats decreased to 55-61% of the control rats. When NDMA was given 6 mg kg by gastric intubation to riboflavin deficient rats, hepatic GSH content decreased markedly to 39-43% of the control rats. After supplying riboflavin, hepatie GSH content of the deficient rats recovered to the level of the control rats. These results suggest that alterations of rat hepatic GSH content during riboflavin deficiency may imply as one of the promoting effects of riboflavin deficiency on the carcinogenesis of nitrosamines.

Biochemical and Morphological Changes in the Lenses of Selenium and/or Vitamin E Deficient Rats

The activities of glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-R),superoalde dismutase (SOD) and the contents of malondialdehyde (MDA) and free radicals were measured, and the morphological changes were observed in the lens of control rats, selenium-deficient (SeD) and/or vitamin E deficient (VED) rats. The activities of GSH-Px in the lens of SeD rats decreased significantly. The GSH-Px activities of lens were positively related to erythrocytes selenium level. There was a free radical at g = 2.0015 in the rat lens of all groupe, but the content of free radicals in the lens of SeD group was significantly higher than that of the control group. The free radical content of lens was negatively related to erythrocytes selenium level, as well as the GSH-Px activities in the lens. In vitro, ultraviolet radiation caused the generation of another kind of free radical (g = 2.0097) in the lens of all groups, but the amount of the free radical in the lens of the SeD group was also significantly higher than that of the control group. The activities of SoD and GSSG-R in VED rat lens were significantly decreased. The amount of MDA in the lens of SeD and/or VED rats were significantly increased. The results showed that the decrease of antioaldative capability in the lenses of SeD and/or VED rats accelerated the lipid peroxidation and generation of free radicals. Although only early morphological changes in SeD and/or VED rat lens were observed, it is considered that selenium and vitamin E deficiency may be involved in the occurrence of cataract

Seasonal climate effect on organ proportion and morbidity in 'Bi-zheng' rats with kidney deficiency syndrome

Objective:Using the traditional Chinese medicine theory of 'Bi-zheng' caused by kidney deficiency,combined with modern medical science and technology,this study assessed the relationships among organ weight,cytokines,pathological manifestations,and traditional Chinese medicine kidney deficiency syndrome in rats with rheumatoid arthritis.Methods:Sixty Sprague-Dawley rats were randomly divided into normal,collagen-induced arthritis (ClA) model,and kidney deficiency arthralgia model groups.Rats in the kidney deficiency arthritis group underwent removal of the testis or ovary and administration of an inflammatory agent.Rats in the control group solely received an inflammatory agent.Type Ⅱ collagen-induced arthritis was induced within 1 week of the winter solstice or summer solstice,in order to sample organs and observe the mental state,posture,fur color,activity level,excrement,body weight,and organ weight of the rats.Results:Compared with the normal and CIA model groups,the kidney deficiency arthralgia model group showed fast and acute onset,prominent symptoms,most obvious potential joint swelling,worst general state,and least amount of weight gain.Compared with the normal group,the proportion of thymus,spleen,and adrenal glands of model rats were larger in winter,with varying degrees of hyperplasia and hypertrophy;these differences were statistically significant (P <.05).In summer,model coefficients were greater than normal in the spleen,while the thymus and adrenal gland coefficients were significantly different between the normal and kidney deficiency arthralgia groups.In summer,the thymus in the kidney deficiency arthralgia group exhibited hypertrophy and hyperplasia,whereas the thymus in the control group did not;this difference was statistically significant (P =.015).Thymus and adrenal gland proportions exhibited seasonal differences in CIA model rats;these were statistically significant (P <.05),with high performance in winter and low performance in summer.The spleen proportion exhibited a trend of high performance in winter and low performance in summer in the kidney deficiency arthralgia model rats.Conclusion:Kidney deficiency is an internal condition linked with the occurrence of arthralgia,and seasonal factors exhibit an impact on the morbidity of arthritis in rats.

Rat Models of Fatty Liver due to Liver Depression and Spleen Deficiency

[Objectives] To establish animal models of fatty liver due to liver depression and spleen deficiency that were suitable for activity discovery of traditional Chinese medicine( TCM) compounds,efficacy evaluation,new drug research and development and correspondence between prescription and syndrome of TCM. [Methods] A syndrome score scale that was suitable for evaluating the rat models was established according to the evolvement rules of etiology and pathogenesis of TCM and the modern clinical pathological mechanism. At the same time,bifendate pills and Sanqi Zhigan pills were selected as drug counterevidence for the models. Rats in model groups were given different proportions of high-fat and low-protein fodder and different concentrations of alcohol every day,and intraperitoneally injected with porcine serum twice a week. Drug groups were given with bifendate pills and Sanqi Zhigan pills( 8. 1 mg/kg,and 2. 7 g/kg) respectively for 14 consecutive days.During the experiment,general status,weight,daily fodder intake and daily water intake of the animals were observed,and TCM syndromes were scored. After the experiment,the levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),γ-glutamine transpeptidase( γ-GT),total cholesterol( TC),triglycerides( TG),low density lipoprotein-cholesterol( LDL-C),and high density lipoprotein-cholesterol( HDL-C) in serum were detected,and the pathological changes in liver tissue were observed. [Results] Compared with the control group,body weight,daily fodder and water intake of rats increased slowly in the model group,and the levels of ALT,AST,γ-GT,TC,TG and LDL-C increased significantly,while the level of HDL-C dropped. Pathological examination showed that steatosis and fat granule were observed in hepatocytes of rats in the model group. Behavioral observation found that main symptoms and minor symptoms of rats in the model group conformed to the syndrome manifestation of liver depression and spleen deficiency,which suggested that the three model groups were established successfully. Among them,three rats died in model group 1,and one rat died in model group 2. The manifestation of all the above lesions can be alleviated by drug counterevidence. [Conclusions]The animal model of fatty liver due to liver depression and spleen deficiency shows an obviously lower mortality and shorter duration and can be used for research of correlation between prescription and syndrome and its mechanism.

Apoptosis Induced by Zinc Deficiency in Rat Osteoblast:Possible Involvement of Protein Kinase C

Ra ostcobasts were isolated from the 21-day fetal rat calvchas. The cells were grown in DMEM Plus 10% FBS, and were treated for 24 h. with 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+. Apoptosis of osteoblasts were measured by fiow cytometry, electron microscopy and DNA fragmentation analyzed by gel elecmphoresis. In addition, IP3 production and PKC activity were measmed in ordr to show whether they are involved in apoptosis in osteoblast induced by alnc deficiency. The results showed that 10 μmol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells ed with 10μmol/L TPEN supplemented with 10 μmol/L Zn2+ showed no apoptotic changs in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supPlemented with Zn2+ simulaneosly and the untreated cells. It can be inferred that apoptosis induced by ainc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.

Discussion on the Relationship Between Senescence Caused by Pi(Spleen)Deficiency and Mitochondrial Autophagy

The Pi(Spleen)is responsible for the formation and transportation,and it is in the center.It is the source of qi and blood generation and transformation,so it has an essential position.If the qi and blood are biochemically passive,the functions of the human body will age prematurely.Mitochondria are for energy synthesis and have always been a hot topic in modern medical research.Many of their functions are similar to those of the Pi in Chinese medicine.In Chinese medicine,strengthening the Pi is to enhance the Pi’s ability to transport and transform,and to speed up the Pi,which is similar to the ability of mitochondria to maintain the ability to remove damaged cells.This can speed up the body’s ability to absorb nutrients and delay aging.

The testosterone mimetic properties of icariin

Aim： To evaluate the testosterone mimetic properties of icariin. Methods： Forty-eight healthy male Sprague-Dawley rats at the age of 15 months were randomly divided into four groups with 12 rats each： the control group （C）, the model group （M）, the icariin group （ICA） and the testosterone group （T）. The reproductive system was damaged by cyclophosphamide （intraperitoneal injection, 20 mg/kg·day） for 5 consecutive days for groups M, ICA and T, at the sixth day, ICA （gastric gavage, 200 mg/kg·day） for the ICA group and sterandryl （subcutaneous injection, 5 rag/rat.day） for the T group for 7 consecutive days, respectively. The levels of serum testosterone, luteinizing hormone （LH）, follicle stimulating hormone （FSH）, serum bone Gla-protein （BGP） and tartrate-resistant acid phosphatase activity in serum （StrACP） were determined. The histological changes of the testis and the penis were observed by microscope with hematoxylin-eosin （HE） staining and terminal deoxynucleotidyl transferase biotin-dUTP-X nick end labeling （TUNEL）, respectively. Results： （1） Icariin improved the condition of reproductive organs and increased the circulating levels of testosterone. （2） Icariin treatment also improved the steady-state serum BGP and might have promoted bone formation. At the same time, it decreased the serum levels of StrACP and might have reduced the bone resorption. （3） Icarrin suppressed the extent of apoptosis of penile cavernosal smooth muscle cells. Conclusion： Icariin has testosterone mimetic properties and has therapeutic potential in the management of hypoandrogenism.

Effects of High Fluoride and Low Iodine on Thyroid Function in Offspring Rats

Thirty-two Wistar rats were divided randomly into four groups of eight with six females and two males in each group. The rats were exposed to high fluoride drinking water （45 mg F-L^-1 from 100 mg NaF L-1）, low dietary iodine （0.0855 mg kg-1）, or both together in order to assess the effects of these three regimens on the thyroid function of the offspring rats. After the animal model was established, the offspring rats were bred and 10-, 20-, 30-, 60-, and 90-d-old rats were used for the experiment. The treatments for the offspring rats were the same as those of their parents. In comparison with control rats, the relative thyroid glands were changed by three regimens, but the mean values of thyroid weight in the experimental groups saw no marked difference. Serum TT3 levels were increased in all stages in the low iodine （LI） group. In the high fluoride （HiF） group, increase in TT3 levels was observed except in 20-d-old rats. Decrease in TT3 at 20- and 90-d and increase in TT3 at 30- and 60-d were found in HiF＋LI group. Serum TT4 levels first saw an increase, and then dropped in the LI and HiF＋LI group. However, an increase in TT4 was found in the HiF group. The levels of TSH in serum rocketed at d 20, and then dropped in the next stages in experimental groups. The results suggested that thyroid disorder could be induced by high flroride in drinking water, low iodine diet, or both of them. Exposure time to fluoride or low iodine diet was one of the important factors that fluoride can induce the development of thyroid dyfunction.

Intestinal microflora molecular markers of spleen-deficient rats and evaluation of traditional Chinese drugs

AIM:To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient(spleen-deficient) rats and to evaluate traditional Chinese drugs.METHODS:Enterobacterial repetitive intergenic consensus-PCR(ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats.RESULTS:A molecular marker was identified for Pi-deficiency in rats.The pharmacodynamic evaluation system,including identified molecular markers(net integral area and abundance of DNA bands),Shannon's index for diversity of intestinal microflora,and Sorenson's pairwise similarity coefficient,was established.The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats,especially at their medium dose(equivalence to the clinical dose),produced more pronounced recovery activities in Pi-deficient rats,while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora.CONCLUSION:Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment.The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.

Baicalin protects neonatal rat brains against hypoxicischemic injury by upregulating glutamate transporter 1 via the phosphoinositide 3-kinase/protein kinase B signaling pathway

Baicalin is a flavonoid compound extracted from Scutellaria baicalensis root.Recent evidence indicates that baicalin is neuroprotective in models of ischemic stroke.Here,we investigate the neuroprotective effect of baicalin in a neonatal rat model of hypoxic-ischemic encephalopathy.Seven-day-old pups underwent left common carotid artery ligation followed by hypoxia（8% oxygen at 37°C） for 2 hours,before being injected with baicalin（120 mg/kg intraperitoneally） and examined 24 hours later.Baicalin effectively reduced cerebral infarct volume and neuronal loss,inhibited apoptosis,and upregulated the expression of p-Akt and glutamate transporter 1.Intracerebroventricular injection of the phosphoinositide 3-kinase/protein kinase B（PI3 K/Akt） inhibitor LY294002 30 minutes before injury blocked the effect of baicalin on p-Akt and glutamate transporter 1,and weakened the associated neuroprotective effect.Our findings provide the first evidence,to our knowledge that baicalin can protect neonatal rat brains against hypoxic-ischemic injury by upregulating glutamate transporter 1 via the PI3 K/Akt signaling pathway.

Beneficial effects of AOS-iron supplementation on intestinal structure and microbiota in IDA rats

The objective of this study was to investigate the effects of agar oligosaccharide-iron(AOS-iron)on intestinal tissue pathology and microbiota in IDA rats induced by a low-iron diet,further to find the relationship between intestinal microbiota and iron metabolic disorders.After 4 weeks of AOS-iron supplementation,the fecal iron content of IDA rats markedly increased in a dose-dependent manner,only the damaged cecum and colon tissues in medium-dose(MD)and high-dose(HD)groups were repaired to the baseline,while the diversity of gut microbiota was improved even at low dose(LD).Furthermore,the supplementation of AOS-iron altered the composition of gut microbiota.At the genus level,the beneficial microbiota was enriched in AOS-iron groups,but the relative abundance of potential opportunistic pathogens obviously reduced compared to that in the anemia model(AM)group.Spearman’s correlation analysis revealed that biochemical parameters,including blood metabolic parameters,iron contents,body weight,GSH-PX and T-AOC activity,were positively correlated with SMB53,Anaerotruncus,Anaerostipes and Coprobacillus but negatively correlated with Morganella,Fusobacterium and Serratia.These findings indicated that AOS-iron effectively repaired the damaged intestinal tissue and ameliorated iron metabolic disorders by regulating gut microbiota desirably,which could provide references for the treatment of IDA.