Search

The present study measured serum levels of vitamin A (VA) and vitamin D (VD) in children with chronic tic disorders (CTD) and investigated their potential association with CTD and comorbidity of attention deficit hyperactivity disorder (ADHD) and the association of their co-insufficiencies or deficiencies with CTD symptoms. A total of 176 children (131 boys and 45 girls, median age of 9 years) with CTD were recruited as the CTD group. During the same period, 154 healthy children were selected as the healthy control (HC) cohort. Circulating retinol and 25-hydroxyvitamin D (25[OH]D) levels were measured for all participants using high-performance liquid chromatography (HPLC) and tandem mass spectrometry. The Yale Global Tic Severity Scale (YGTSS) was employed for the assessment of tic status and CTD impairment. The Swanson, Nolan, and Pelham Rating Scale (SNAP-IV) and the Children’s Yale-Brown Obsessive–Compulsive Scale (CY-BOCS) were used to evaluate comorbidity symptoms. CTD pediatric participants exhibited markedly diminished circulating retinol and 25(OH)D levels compared to HCs. Moreover, VA and VD deficiencies and their co-insufficiencies/deficiencies were more prevalent in CTD participants than HCs. Circulating 25(OH)D levels were inversely proportional to the YGTSS motor tic scores. YGTSS scores in CTD children with only VA or VD insufficiency or deficiency or with VA and VD co-insufficiency/deficiency did not differ from those in CTD children with normal VA and VD. CTD children with comorbid ADHD displayed reduced circulating retinol and 25(OH)D concentrations and elevated prevalence of VD deficiency compared to CTD participants without comorbid ADHD. Lower serum retinol content was intricately linked to the presence of elevated CTD and comorbid ADHD. VA and VD deficiencies and their co-insufficiencies/deficiencies were markedly enhanced in CTD pediatric participants compared to HCs. Lower VA concentration was linked to the presence of enhanced CTD and comorbid ADHD. Therefore, children with CTD, especially with comorbid ADHD, may be at a higher risk of VA or VD deficiency, which may prompt the clinicians to consider whether blood tests for VA and VD in CTD children would be helpful for clinical care.

Purpose Vitamin D deficiency has been found in children with chronic tic disorders (CTDs). Our previous data showed that serum 25-hydroxyvitamin D [25(OH)D] level in children with CTDs was lower than that of the healthy controls and lower serum 25(OH)D level was associated with increased severity of the tic disorder. Thus, we intend to further verify this phenomenon and examine the effect of vitamin D3 on CTDs. Patients and methods In total, 120 children with CTDs and 140 normal controls were enrolled in this study, with 36/120 of those in the CTD group receiving vitamin D3 treatment for 3 months. The Yale Global Tic Severity Scale (YGTSS) and Clinical Global Impression of Severity of Illness (CGI-SI) were, respectively, used to evaluate the tic severity. High-performance liquid chromatography and tandem mass spectrometry were used to measure serum 25(OH)D level. Results Those children with CTDs exhibited significantly lower 25(OH)D levels than did healthy controls, and these reduced 25(OH)D levels were linked to increasing severity of tic symptoms. After treatment with supplemental vitamin D3, serum 25(OH)D level and scores of YGTSS total, motor tics, phonic tics, total tic, impairment, and CGI-SI improved significantly in children with CTDs without any adverse reactions. Conclusion Supplementation vitamin D3, given its low cost and excellent safety, may be an effective means of improving symptoms in certain children with CTDs.

Background: Electronic screen media play an increasingly vital role in children's entertainment; however, excessive screen time may negatively influence child development. The purpose of this study was to investigate the relationship between the screen time of children with autism spectrum disorder (ASD) and their autistic symptoms and development quotients (DQs).Methods: We compared the screen time of 101 children with ASD and 57 typically developing (TD) children. Then, we performed a correlation analysis to determine the correlations between the screen time and the ASD-related scale scores and developmental quotients of the Gesell Developmental Schedules (GDS) of ASD children. We further divided the ASD group into subgroups according to the screen time and age and then separately conducted the above correlation analyses by subgroup.Result: The results showed that the screen time of the children with ASD was longer than that of the TD children (3.34 ± 2.64 h vs. 0.91 ± 0.93 h). The screen time of the children with ASD was positively correlated with the Childhood Autism Rating Scale (CARS) score (r = 0.242, P = 0.021) and “taste, smell and touch” item of CARS(r = 0.304, P = 0.005), and negatively correlated with the language DQ of the GDS (r = −0.236, P = 0.047). The subgroup analysis showed that in the longer screen time subgroup of ASD children, the screen time was positively correlated with the CARS score (r = 0.355, P = 0.026) and negatively correlated with the DQs of all domains of the GDS (P < 0.05). In addition, in the younger age group of ASD children, the screen time was positively correlated with the CARS score (r = 0.314, P = 0.021) and negatively correlated with the DQs of all domains of the GDS, except for the personal-social behavior domain (P < 0.05).Conclusion: Compared with TD children, children with ASD have a longer screen time. The screen time is related to autism-like symptoms and the DQs of children with ASD. The longer the screen time, the more severe the symptoms of ASD (especially sensory symptoms), and the more obvious the developmental delay, especially in ASD children with a longer screen time and younger age, particularly in the language domain.

Inflammatory bowel disease (IBD) is an intestinal chronic inflammatory disease, and is related to imbalance of CD4+T subsets. However, the current treatments of chronic colitis are not ideal and have potential side effects. Therefore, more effective and safer biologically active substances which are extracted from natural plants have been widely concerned. In this study, it was found that Inonotus obliquus polysaccharides (IOP), the main bioactive constituent of Inonotus obliquus, can alleviate dextran sodium sulfate-induced chronic murine intestinal inflammation. Oral administration of IOP (100, 200, 300 mg/kg) can significantly reduce the disease active index and alleviate the pathological changes in colitis mice, where the tight junction proteins Occludin and ZO-1 losses in colon tissues were reduced. It can also regulate imbalanced Th1/Th2 and Th17/Treg in colon tissues, mesenteric lymph nodes and spleen using Reverse Transcription-Polymerase Chain Reaction detection and flow cytometry. Immunohistochemistry and western blot assays further revealed the modulatory effect of IOP on the p-STAT1, p-STAT6, p-STAT3 expression, which promoted the balance of Th1/Th2, Th17/Treg in the colon of chronic colitis mice. In short, these results indicated that IOP was potentially effective therapeutic agent for IBD.

Background Exposure to phthalate esters may be linked to the risk of high blood pressure (HBP), but limited evidence is available in Chinese children. Objective To investigate the associations between nine phthalate metabolites (mPAEs) and systolic/diastolic BP, pulse pressure (PP), mean arterial pressure (MAP), and the risk of HBP. Methods In this cross-sectional study, a total of 1044 primary school children (6–8 years old) were enrolled from Shenzhen, China, between 2016 and 2017. Nine mPAEs were analyzed from urine using ultra-performance liquid chromatography and tandem mass spectrometry. A multivariable linear regression model was used to explore the associations between phthalate exposure and systolic/diastolic BP, PP, and MAP. A binary logistic regression model was used to examine the associations between phthalate exposure and the risk of HBP. Results Monomethyl phthalate (MMP) concentrations were significantly higher in HBP children than normal BP children. MMP, monoisobutyl phthalate (MiBP), monobutyl phthalate (MnBP), mono(5-carboxy-2-ethylpentyl) phthalate, mono-[(2-carboxy methyl)hexyl] phthalate (MCMHP), the sum of four short-chain mPAEs (∑LMW), and the sum of all nine mPAEs (∑9mPAEs) were significantly positively associated with increases in systolic BP z-score, while only MMP was significantly positively associated with diastolic BP z-score. MMP, MiBP, MnBP, MCMHP, ∑LMW, and ∑9mPAEs were significantly associated with increases in PP, while MMP and MnBP were significantly associated with increases in MAP. MMP was significantly associated with the risk of HBP, with an odds ratio of 1.87 (95% CI: 1.23, 2.85). Conclusions The present study suggests that dimethyl phthalate exposure increases the risk of HBP. And some types of phthalates are associated with elevations in systolic/diastolic BP z scores, PP, and MAP in Chinese children.

The implication of different dietary n-3/n-6 polyunsaturated fatty acids (PUFAs) ratios has been investigated in some neurodevelopmental disorders (including autism and depression). However, the mechanisms underlying the effects of different PUFAs ratios on the autism are still poorly understood. In the present study, a valproic acid (VPA) rat model of autism was used to study the effects of diet with different n-3/n-6 PUFA ratios on the autism, and the underlying mechanisms explored. Our results showed that rats with prenatal administration of VPA took less response time to sniff three odorants in the olfactory habituation/dishabituation tests, had lower frequency of pinning and following patterns, and had decreased hippocampal 5-hydroxytryptamine (5-HT), increased serum 5-HT and downregulated expression of tight junction protein (occludin and claudin-1) in the colon. However, supplementation of n-3/n-6 PUFAs (1:5) in the VPA treated rats ameliorated the autistic behaviors, increased hippocampal 5-HT and tight junction expression in the colon, and decreased serum 5-HT. In conclusion, dietary supplementation of n-3/n-6 PUFAs (1:5) significantly improves VPA-induced autism-like behaviors in rats, which may be, at least partially, related to the increased hippocampal 5-HT. Furthermore, this diet can increase the expression of tight junction proteins to improve intestinal barrier impairment.

Autism spectrum disorder (ASD) is a developmental disability which may cause significant social, communication, and behavioral challenges. Besides certain essential symptoms, a lot of ASD individuals also suffer the comorbidity of gut microbiota dysbiosis, which possibly causes a variety of gastrointestinal (GI) difficulties. Interestingly, evidence has indicated that behavioral output may be modulated through the communication between the central nervous system and gut microbiota via the gut-brain axis. Polyunsaturated fatty acids (PUFAs) and n-3 fatty acids (n-3 PUFA) are structurally and functionally crucial components for the brain, and the state of n-3 PUFAs also affects the gut microbiota. However, how varying intake ratios of n-3/n6 PUFAs affect the gut microbiota composition in ASDs is not well-understood. Pregnant female Wistar rats with intraperitoneal administration of valproate acid (VPA) at embryonic day (E) 12.5 and their male offspring were grouped and fed three diets: a control chow (VPA group), omega-3 deficient (A group), and n-3/n6 (1:5) diet (B group). The diet of pregnant female Wistar rats with intraperitoneal administration of saline and their male offspring was a control chow (normal group). Microbial composition and species abundance were investigated accordingly by the 16S rRNA gene-based metagenomics analysis on the fecal samples. Results showed that fecal microbial abundance was decreased because of VPA administration in the period of pregnancy, and the changing pattern of gut microbiota was similar to that reported in ASD patients. Furthermore, the n-3/n6 (1:5) diet increased the fecal microbial abundance and decreased the elevated Firmicutes. In conclusion, n-3/n6 PUFAs (1:5) diet supplementation may alter gut microbiota composition in VPA-exposed rats. This study put forward a new strategy for the intervention and treatment of autism by n-3/n-6 PUFAs ratio supplementation intakes.

OBJECTIVE: To explore the effect of parental training based on the Early Start Denver Model (ESDM) combined with intensive training on the treatment outcome of children with autism spectrum disorder (ASD) and its impact on parenting stress. METHODS: Seventy children aged 2-5 years who were diagnosed with ASD were enrolled in the study. They were divided into an ESDM group and a parental training group by the random number table method (n=35 each). The ESDM group received intensive training based on ESDM. In addition to intensive ESDM-based training, parents of the children in the parental training group received ESDM skills training. Both groups were assessed by Autism Behavior Checklist (ABC), Childhood Autism Rating Scale (CARS), Autism Treatment Evaluation Checklist (ATEC) and Parenting Stress Index-Short Form (PSI-SF) before and after the intervention of 3 months. RESULTS: After 3 months of intervention, the total scores of ABC, CARS and ATEC were both significantly decreased in the two groups (P < 0.05). There was no significant difference in the total scores of ABC, CARS and ATEC between the two groups before and after intervention (P > 0.05). The change between ABC, CARS and ATEC total scores in the two groups had no significant difference (P > 0.05). After 3 months of intervention, the total scores of PSI-SF were both significantly decreased in the two groups (P < 0.05). The difficult child sub-scale scores in PSI-SF were significantly decreased in the ESDM group (P < 0.05). While three sub-scale scores of parent distress, parent-child dysfunctional interaction and difficult child in PSI-SF were significantly decreased in the parental training group (P < 0.05). Before and after intervention of 3 months, no significant difference was found in PSI-SF total scores between the two groups. Compared with the ESDM group, the change between PSI-SF total scores and two sub-scales of PSI-SF (parent distress and difficult child) were significantly bigger in the parental training group (P < 0.05). CONCLUSIONS: Both the combination of intensive training and parent training based on ESDM and ESDM intensive training alone can improve the core symptoms of children with ASD aged 2-5 years and relieve the parenting stress, however, the former is more effective in relieving parenting stress.

Rationale: Autism spectrum disorder (ASD) is a common neurodevelopmental disorder caused by complex interactions between genetic and environmental factors. Recent studies suggest that Vitamin D3 or bumetanide therapy may improve the core symptoms of ASD in some individuals. However, there are no guidelines that provide clinicians with evidence-based treatment regimens for the use of these therapies in ASD. Patient concerns: A 30-month-old female was referred to our department because she did not respond when her name was called. Diagnosis: The patient was diagnosed with ASD by a team of autism experts according to American Psychiatric Association Diagnostic and Statistical Manual of Mental Disorders (DSM-5) criteria. Interventions: The patient was administered Vitamin D3 150,000 IU intramuscularly once a month and Vitamin D3 800 IU orally each day. After 6 months, Vitamin D3 supplementation was discontinued because of lack of effectiveness. Subsequently, oral bumetanide 0.5 mg twice daily was initiated. Outcomes: The patient's symptoms remained unchanged after 6 months of Vitamin D3 supplementation, and her serum 25 (OH) D levels had reached 52.4 ng/mL. At the parent's request, Vitamin D3 supplementation was discontinued because of lack of effectiveness. Thereafter, bumetanide was initiated. After 1 month of bumetanide, the patient's Childhood Autism Rating Scale score was 26, which is below the cutoff score for ASD. This case report suggests that Vitamin D3 and bumetanide target different mechanisms in the pathogenesis of ASD. Lessons: Based on these observations, we discuss three possible scenarios for Vitamin D3 supplementation and propose that bumetanide should be initiated if Vitamin D3 supplementation is ineffective (identifier ChiCTR-CCC-13004498).

Unmanipulated haploidentical peripheral blood stem cell transplantation (haplo-PBSCT) has been an established treatment to cure high-risk leukemia/lymphoma. Relapse is the main cause of treatment failure for patients with relapsed/refractory disease or with very high-risk gene mutations such as TP53, TET2, and DNMT3a. In this study, we aimed to establish the tolerance and efficacy of prophylactic donor lymphocyte infusion (DLI) with G-CSF-primed peripheral blood progenitors for prevention of relapse in these very high-risk patients after haplo-PBSCT. The prophylactic DLI was given at a median of 77 days after transplantation in 31 of 45 consecutive patients with very high-risk leukemia/lymphoma. The median dose of CD3+ cells for infusion was 1.8 × 107/kg. The 100-day incidences of acute graft-versus-host disease (GVHD) grades 2–4 and 3–4 after DLI were 55.3% and 10.2%. The 2-year incidences of chronic GVHD and severe chronic GVHD were 52.0% and 18.2%. The 2-year incidences of non-relapse mortality and relapse were 33.1% and 32.5%. The 2-year probabilities of overall survival and relapse-free survival were 40.1% and 31.9%. Poor-risk gene mutations (p = 0.029), disease in non-remission status prior to transplantation (p = 0.005), and donors older than 40 years of age (p = 0.043) were associated with relapse after DLI. In multivariate analysis, disease in non-remission status prior to transplantation was an independent risk factor of relapse (hazard ratio = 4.079; p = 0.035). These data showed the feasibility of the prophylactic DLI in the haplo-PBSCT setting and the anti-leukemic efficacy in very high-risk leukemia/lymphoma. Electronic supplementary material The online version of this article (10.1007/s00277-018-3482-7) contains supplementary material, which is available to authorized users.

Hong-Hua Li,1 Jun-Yan Feng,1 Bing Wang,1 Yu Zhang,1 Cheng-Xin Wang,1 Fei-Yong Jia1,2 1Department of Developmental and Behavioral Pediatrics, The First Hospital of Jilin University, Changchun, Jilin 130021, People’s Republic of China; 2Neurological Research Center of the First Hospital of Jilin University, Changchun, People’s Republic of ChinaCorrespondence: Fei-Yong JiaDepartment of Developmental and Behavioral Pediatrics, The First Hospital of Jilin University, Changchun, Jilin 130021, People’s Republic of ChinaTel +86-0431-88783846Email erkekangfujia@163.comBackground: The newly revised Children Neuropsychological and Behavior Scale (CNBS-R2016) is a diagnostic assessment tool widely used in China to assess the developmental level of children aged 0 to 6 years. The purpose of this study was to determine whether the effectiveness of developmental assessment in children with autism spectrum disorder (ASD) by the CNBS-R2016 was consistent with that of the Griffiths Mental Development Scales for China (GDS-C).Methods: In total, 139 children with ASD were recruited in this study. The Autism Behavior Checklist (ABC) and the Childhood Autism Rating Scale (CARS) were used to measure ASD severity. All subjects were evaluated with both the CNBS-R2016 and GDS-C. To determine the consistency between the CNBS-R2016 and GDS-C, Pearson correlation coefficients and Bland-Altman plots were computed. The GDS-C was used as a reference assessment, and the performance of the CNBS-R2016 was analyzed with receiver operating curves.Results: No significant difference was found between the proportions of developmental delays detected by the CNBS-R2016 subscales and the corresponding GDS-C subscales. The CNBS-R2016 Communication Warning Behavior subscale quotients and the total ABC and CARS scores were significantly and positively correlated. The general and subscale quotients of the CNBS-R2016 and the corresponding quotient of the GDS-C were also significantly and positively correlated. The area under all the curves of the CNBS-R2016 was above 0.8 according to the results of the GDS-C (general or subscale quotient

BACKGROUND There is currently little information on haploidentical hematopoietic cell transplantation (haplo-HCT) for T-lymphoblastic lymphoma (T-LBL). Data about peripheral blood stem cells (PBSC) as a reliable graft source for T-LBL treatment are lacking. MATERIAL AND METHODS T-LBL patients who underwent T cell-replete haploidentical peripheral blood hematopoietic cell transplantation (haplo-PBHCT) from July 2007 to January 2017 were retrospectively evaluated. RESULTS A total of 25 patients (age ≥15 years) with median age of 24 (range 15-51) years were enrolled. The median number of CD34+ cells infused was 5.0 (1.6-14.4) 106/kg. Sustained myeloid engraftment with full donor chimerism was achieved in all patients. The cumulative incidence of grades 2 to 4 acute graft-versus-host disease (GVHD) at day 100 was 24%. Two-year extensive chronic GVHD cumulative incidence was 20%. The 3-year overall survival rate for all patients was 70%. The median survival time of the complete remission (CR) group was better than that of the non-CR group (not reached vs. 9 m) (P

In the present study, the gastroprotective effects of arctigenin of Fructus Arctii were evaluated and the possible underlying mechanisms of action were elucidated. Arctigenin (high-performance liquid chromatography purity, >99.0%) was isolated and purified from the seeds of Arctium lappa L. The anti-ulcerogenic activity of arctigenin against ulcers induced by absolute ethanol and acetic acid was evaluated in a Sprague-Dawley rat model. In addition, the antioxidant activity was assessed by measuring malondialdehyde (MDA) levels in an ethanol-induced model and the anti-inflammatory effects were assessed by measuring five factors in an acetic acid-induced model. In the ethanol-induced model, arctigenin inhibited gastric lesions in a dose-dependent manner, by 53.04, 53.91 and 64.43% at doses of 0.05, 0.15 and 0.45 mg/kg, respectively. In addition, arctigenin reduced MDA (P

The development of invasive mechanical ventilation technology provides effective respiratory support for critically ill children. However, respiratory support is not the end of treatment as the ultimate goal is successful extubation in children. At present, some evaluation indicators before extubation including rapid shallow breathing index, maximal inspiratory pressure, and work of breathing are of high clinical value in predicting adult extubation outcome, but their evidence of evidence-based medicine is not sufficient in the field of pediatric intensive care. This paper reviews the current research on the validity of predictors for extubation outcomes in children. It shows that there is still a lack of indicators with good sensitivity and specificity for assessment before extubation in children. The studies are still in a small-sample size and single-center stage. Therefore, how to optimize evaluation before extubation and improve the success rate of extubation is the direction of joint efforts of doctors in the pediatric intensive care unit and rehabilitation medicine department.

Liping Dou,1,2* Qingyu Xu,1* Mengzhen Wang,1* Yang Xiao,1* Longcan Cheng,2 Honghua Li,1 Wenrong Huang,1 Junhui Mei,1 Yu Jing,1 Jian Bo,1,2 Daihong Liu,1 Li Yu11Department of Hematology, Chinese PLA General Hospital, Beijing 100853, People’s Republic of China; 2Department of Hematology, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan 572013, People’s Republic of China*These authors contributed equally to this work Purpose: The chemotherapeutic regimen DCAG (decitabine with cytarabine, aclarubicin hydrochloride, and granulocyte colony-stimulating factor) is effective for elderly patients with acute myeloid leukemia, but recommendations for young patients remain controversial. This study investigated the tolerance and efficacy of DCAG for patients with newly diagnosed acute myeloid leukemia (aged 14–60 years). The clinical features or molecular markers that may predict response to DCAG were identified.Patients and methods: One-hundred sixty-one consecutive patients with newly diagnosed acute myelogenous leukemia received DCAG or standard (idarubicin plus cytarabine, IA) induction chemotherapy (n=64 and 97, respectively).Results: The rates of complete remission after the first cycle, overall survival (OS), and event-free survival (EFS) were comparable. After the second cycle, the complete remission rate of the DCAG group (54.7%) was significantly lower than that of the reference (78.35%, P=0.005). The following were associated with significantly worse OS, and EFS, in the DCAG group: Eastern Cooperative Oncology Group (ECOG) score ≥3 and no response after the second induction therapy; and FLT3-ITD. The multivariate analysis showed the DCAG group with significantly shorter OS associated with ECOG ≥3 and FLT3-ITD. In the DCAG group, after the first cycle of induction chemotherapy the median recovery times of neutrophils and platelets were 15.8 and 13 days.Conclusion: The DCAG and IA groups were similar with regard to complete remission rate after the first cycle, OS, and EFS. The complete remission rate after the second cycle of the DCAG was significantly lower than that of the IA. Grade 4 neutropenia and thrombocytopenia were a major adverse event associated with DCAG.Keywords: decitabine, acute myeloid leukemia, induction therapy, conventional chemotherapy

OBJECTIVE: To study the clinical effect of vitamin D(3) (VitD(3)) combined with the Early Start Denver Model (ESDM) in the treatment of autism spectrum disorder (ASD) in toddlers. METHODS: A total of 102 toddlers with ASD, aged 1 to 3 years, were enrolled. According to the wishes of their parents, they were divided into conventional rehabilitation, ESDM and ESDM+VitD(3) groups. Autism Behavior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were used evaluate behavior problems before treatment and after 3 months of treatment. RESULTS: The conventional rehabilitation group had significant reductions in the total score and the scores on somatic movement and self-care subscales of the ABC scale after 3 months of treatment (P < 0.05). After 3 months of treatment, the ESDM group had significant reductions in the total score and the scores on somatic movement, self-care, social interaction and language subscales of the ABC scale (P < 0.05), as well as a significant reduction in the total score of the CARS (P < 0.05). After 3 months of treatment, the ESDM+VitD(3) group had a significant increase in the level of 25(OH)D and significant reductions in the total score and the scores on self-care, sensation, social interaction and language subscales of the ABC scale (P < 0.05), as well as a significant reduction in the total score of the CARS (P < 0.05). The ESDM group had a significantly greater reduction in the score on social interaction subscale than the conventional rehabilitation group (P < 0.05). The ESDM+VitD(3) group had a significantly greater reduction in the score on social interaction subscale than the other two groups (P < 0.05). CONCLUSIONS: ESDM can effectively improve the clinical symptoms of toddlers with ASD, with a significantly better clinical effect in improving social interaction and somatic movement than conventional rehabilitation. ESDM combined with VitD(3) has a significantly better clinical effect in improving social communication skills and may be one of the best strategies for improving the clinical symptoms of toddlers with ASD.

Tic disorders (TD) are a group of neurodevelopmental disorders that are characterized by motor and/or vocal tics in children and adolescents. The etiology and pathogenesis of TD remain unclear, and it is believed to be caused by a combination of genetic, biological, psychological, and environmental factors. The major treatment for TD includes psychoeducation, behavioral intervention, and drug treatment. To further explore the management of TD, this article reviews the research advances in psychoeducation and behavioral intervention for patients with TD.

Background Both type 2 diabetes mellitus (T2DM) and Alzheimer's disease (AD) are common age-associated disorders and T2DM patients show an increased risk to suffer from AD, however, there is currently no marker to identify who in T2DM populations will develop AD. Since glycogen synthase kinase-3β (GSK-3β) activity, ApoE genotypes and olfactory function are involved in both T2DM and AD pathogenesis, we investigate whether alterations of these factors can identify cognitive impairment in T2DM patients. Methods The cognitive ability was evaluated using Minimum Mental State Examination (MMSE) and Clinical Dementia Rating (CDR), and the mild cognitive impairment (MCI) was diagnosed by Petersen's criteria. GSK-3β activity in platelet, ApoE genotypes in leucocytes and the olfactory function were detected by Western/dot blotting, the amplification refractory mutation system (ARMS) PCR and the Connecticut Chemosensory Clinical Research Center (CCCRC) test, respectively. The odds ratio (OR) and 95% confidence intervals (95% CI) of the biomarkers for MCI diagnosis were calculated by logistic regression. The diagnostic capability of the biomarkers was evaluated by receiver operating characteristics (ROC) analyses. Findings We recruited 694 T2DM patients from Jan. 2012 to May. 2015 in 5 hospitals (Wuhan), and 646 of them met the inclusion criteria and were included in this study. 345 patients in 2 hospitals were assigned to the training set, and 301 patients in another 3 hospitals assigned to the validation set. Patients in each set were randomly divided into two groups: T2DM without MCI (termed T2DM-nMCI) or with MCI (termed T2DM-MCI). There were no significant differences for sex, T2DM years, hypertension, hyperlipidemia, coronary disease, complications, insulin treatment, HbA1c, ApoE ε2, ApoE ε3, tGSK3β and pS9GSK3β between the two groups. Compared with the T2DM-nMCI group, T2DM-MCI group showed lower MMSE score with older age, ApoE ε4 allele, higher olfactory score and higher rGSK-3β (ratio of total GSK-3β to Ser9-phosphorylated GSK-3β) in the training set and the validation set. The OR values of age, ApoE ε4 gene, olfactory score and rGSK-3β were 1.09, 2.09, 1.51, 10.08 in the training set, and 1.06, 2.67, 1.47, 7.19 in the validation set, respectively. The diagnostic accuracy of age, ApoE ε4 gene, olfactory score and rGSK-3β were 0.76, 0.72, 0.66, 0.79 in the training set, and 0.70, 0.68, 0.73, 0.79 in the validation set, respectively. These four combined biomarkers had the area under the curve (AUC) of 82% and 86%, diagnostic accuracy of 83% and 81% in the training set and the validation set, respectively. Interpretation Aging, activation of peripheral circulating GSK-3β, expression of ApoE ε4 and increase of olfactory score are diagnostic for the mild cognitive impairment in T2DM patients, and combination of these biomarkers can improve the diagnostic accuracy., Highlights • ApoE ε4 gene, platelet GSK-3β activation, olfactory dysfunction and aging are non-invasive, affordable and accessible biomarkers for diagnosing mild cognitive impairment in type 2 diabetes mellitus patients, and the combination of these non-invasive, affordable and accessible biomarkers can improve the accuracy of the diagnosis. Epidemiological studies show that type 2 diabetes mellitus is an independent risk factor of Alzheimer disease, and a large proportion of diabetic patients will develop Alzheimer disease, but no early diagnostic tool to identify them. We find that ApoE ε4 gene, platelet GSK-3β activation, olfactory dysfunction and aging are early markers for dementia in type 2 diabetes patients, and combination of these non-invasive markers can improve the diagnostic accuracy. These findings shed light on the early identification in type 2 diabetes population who will develop Alzheimer disease and thus enable early intervention to this currently incurable neurodegenerative disorder.

Intermediate-risk acute myeloid leukemia (IR-AML), which accounts for a substantial number of AML cases, is highly heterogeneous. Although several mutations have been identified, the heterogeneity of AML is uncertain because novel mutations have yet to be discovered. Here we applied next generation sequencing (NGS) platform to screen mutational hotspots in 410 genes relevant to hematological malignancy. IR-AML samples (N=95) were sequenced by Illumina Hiseq and mutations in 101 genes were identified. Only seven genes (CEBPA, NPM1, DNMT3A, FLT3-ITD, NRAS, IDH2 and WT1) were mutated in more than 10% of patients. Genetic interaction analysis identified several cooperative and exclusive patterns of overlapping mutations. Mutational analysis indicated some correlation between genotype and phenotype. FLT3-ITD mutations were identified as independent factors of poor prognosis, while CEBPA mutations were independent favorable factors. Co-occurrence of FLT3-ITD, NPM1 and DNMT3A mutations was identified with associated with specific clinical AML features and poor outcomes. Furthermore, by integrating multiple mutations in the survival analysis, 95 IR-AML patients could be stratified into three distinct risk groups allowing reductions in IR-AML by one-third. Our study offers deep insights into the molecular pathogenesis and biology of AML and indicated that the prognosis of IR-AML could be further stratified by different mutation combinations which may direct future treatment intervention.

Preterm birth is a major factor which induces neurological and motor impairments, particularly cerebral palsy, in high-risk infants. Early identification of potential neurodevelopmental impairments provides the opportunity to improve neurodevelopmental outcomes in preterm infants through early rehabilitation interventions. Clinically, the general movement assessment is a pivotal tool to predict neurodevelopmental outcomes, especially motor developmental outcomes, in high-risk infants. Movement recognition can continuously capture relevant limb movements and perform objective and quantitative assessment using computerized approaches. Various methods of recording and analyzing spontaneous general movements for infants at a risk of cerebral palsy have been extensively explored. This article summarizes the general movement assessment method and reviews the translational research on using movement recognition technology for the assessment of spontaneous general movements of preterm infants.

Background Post-transplant lymphoproliferative disease (PTLD) is a rare and serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) or solid organ transplantation. Material/methods We conducted a retrospective analysis of the occurrence of post-transplant lymphoproliferative disease in allo-HSCT recipients over 12 years in a single center in China. A total of 343 patients received allo-HSCT. The conditioning therapy consisted of a busulfan/cyclophosphamide-based regimen, a fludarabine/cyclophosphamide-based regimen, or total-body irradiation and cyclophosphamide. In transplantations from unrelated donors and haplo-identical donors, patients also received antithymocyte globulin (ATG) or thymoglobulin as part of the conditioning. Results Five of the 343 patients (1.46%) were diagnosed with PTLD and all 5 were given ATG as part of conditioning. Among these 5 patients, 4 had lymphoid neoplasm before transplantation. EBV-positivity was confirmed in 4 patients. All 5 PTLD patients received reduction of immunosuppression (RI) as fundamental therapy. At follow-up on April 1, 2013, 1 patient had survived for 2 years and 1 had survived for 9 years. The correlation of PTLD with ATG and underlying diseases were examined by statistical analysis using the chi-squared test or Fisher's exact test (P=0.011 and 0.025, respectively). Conclusions Although only 1.46% of patients progressed to PTLD associated with ATG and underlying diseases, the mortality was still high. Moreover, RI can be an effective therapy for PTLD patients, but other approaches should be further explored.

Rationale West syndrome (WS) is an age-dependent epileptic encephalopathy that is characterized by intractable epileptic seizures, hypsarrhythmia, and observed through electroencephalogram (EEG) and significant neurodevelopmental regression. The spontaneous remission of epileptic seizure is clinically rare and has not previously been reported in a Chinese infant. Herein, we reported a Chinese infant with WS whose seizures disappeared following a human herpesvirus 7 (HHV-7) infection. Patient concerns The male Chinese infant was born at the gestational age of 36 weeks with a birth weight of 1.65 kg and an Apgar score of 7 at the first minute. At the age of 6 months, the infant developed seizures that manifested as flexor spasms with trunk involvement and mental regression. Diagnosis Brain magnetic resonance imaging revealed leukomalacia of the posterior horn and a reduction in the size of the periventricular of the bilateral ventricle and the corpus callosum. An EEG revealed hypsarrhythmia and typical spasm seizures. Therefore, the infant was diagnosed with symptomatic WS. Interventions The infant was treated with adequate vitamin B6 intravenous drip and oral treatment with topiramate and levetiracetam. Outcomes The observed seizures disappeared spontaneously 40 days after onset, without any changes in the anti-epileptic drug treatment, following a febrile rash due to a HHV-7 infection. Lessons Spontaneous remission of epileptic seizures can occur following viral infection of HHV-7 in children with WS. The mechanism behind this spontaneous remission warrants further research.

In the present study, the gastroprotective effects of arctigenin of Fructus Arctii were evaluated and the possible underlying mechanisms of action were elucidated. Arctigenin (high-performance liquid chromatography purity, >99.0%) was isolated and purified from the seeds of Arctium lappa L. The anti-ulcerogenic activity of arctigenin against ulcers induced by absolute ethanol and acetic acid was evaluated in a Sprague-Dawley rat model. In addition, the antioxidant activity was assessed by measuring malondialdehyde (MDA) levels in an ethanol-induced model and the anti-inflammatory effects were assessed by measuring five factors in an acetic acid-induced model. In the ethanol-induced model, arctigenin inhibited gastric lesions in a dose-dependent manner, by 53.04, 53.91 and 64.43% at doses of 0.05, 0.15 and 0.45 mg/kg, respectively. In addition, arctigenin reduced MDA (P

Emerging data demonstrate promising results of related haploidentical allogeneic hematopoietic stem cell transplantation (haplo-HCT) for the treatment of hematologic malignancies. Data about G-CSF primed PBSC as reliable source of graft with myeloablative conditioning are lacking. We updated the outcomes in 130 adult patients with high-risk hematologic malignancies who received haplo-PBSC transplantation consecutively under busulfan-based conditioning. PBSC were freshly isolated and infused without ex vivo T-cell depletion into the recipients. Myeloid recovery was achieved in 99.2% patients with full donor chimerism. The cumulative incidence of acute grade 3-4 GvHD, overall and extensive chronic GvHD was 14.9%, 38.6% and 16.5%, respectively. The 3-year non-relapse mortality rate was 24.1%. Non-remission prior to transplant was associated with higher incidence of relapse (P=0.006), inferior overall survival (P=0.017) and leukemia-free survival (P=0.024). These data suggest that PBSC is a reliable graft source in haploidentical, unmanipulated transplant settings under myeloablative conditioning in patients with high-risk hematologic malignancies.

Background Subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is an uncommon subtype of peripheral T-cell lymphoma and its standard treatment remains ongoing study. Cases refractory to chemotherapy are suggested to benefit from autologous hematopoietic stem cell transplantation (auto-HSCT). Nevertheless, post auto-HSCT recurrence seems to be irreversible. Very few cases of allogeneic-HSCT (allo-HSCT) in SPTCL have been reported so far. Case report A 27-year-old female presented to our hospital with refractory SPTCL for receiving auto-HSCT. Cutaneous biopsy had confirmed diagnosis of SPTCL at a local hospital, where 5 lines and 10 cycles of intensive chemotherapy had been prescribed. All these treatment provided marginal control of disease progression. After her enrollment at our institute, she relapsed again after the collection of peripheral hematopoietic stem cells. Then, we performed an allo-HSCT to her from a 10/10 matched unrelated donor. Neutrophil and platelet engraftment occurred at day +11 and +13, respectively. At day +15, lymphoma, however, came back, resulting in the withdrawal of cyclosporine from day +35 to day +75, when acute graft versus host disease (aGvHD) emerged. Lymphoma cells were observed in a subcutaneous biopsy from her leg. Eventually, skin manifestation of extensive chronic GvHD developed. By present, the patient has sustained complete remission for 18 months. Conclusions Allo-HSCT is rational for patients with refractory SPTCL. To our knowledge, this is the first case, with regained remission after post allo-HSCT relapse through cyclosporine withdrawal, showing the existence of graft versus lymphoma (GvL) effect in SPTCL.

The aim of this study is to evaluate serum 25-hydroxyvitamin D[25(OH)D] levels in children with tic disorders and to explore the relationship between serum 25(OH)D level and tic severity. Children (n = 179, 31 females, 148 males, mean age at diagnosis: 8.0 ± 2.7 years old, age ranged from 3 to 14.5 years old) who were diagnosed with a tic disorder were enrolled as case group, 189 healthy children were recruited as control group. Serum level of 25(OH)D of each child was measured by high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS). Yale Global Tic Severity Scale (YGTSS) was used to assess tic severity. Mean serum level of 25(OH)D in the case group was significantly lower than that of the control group. The serum 25(OH)D level was significantly associated with tic severity after adjusting for age and body mass index (BMI). This study identified a high prevalence of vitamin D insufficiency or deficiency in children with tic disorders, and there was a negative correlation between the serum 25(OH)D level and tic severity. In the future, large sample size studies are urgently needed to further clarify this correlation.

Background and objectives: Newborns with perinatal risk factors and abnormal general motor quality assessment are at high risk of developing cerebral palsy. The treatment is earlier, the prognosis is better. This trial will investigate whether, combined with conventional early interventional therapy, the use of mouse nerve growth factor in high-risk infants can improve their motor and cognitive abilities. Design: This is a prospective, single-center, randomized, parallel, controlled, clinical trial. Methods: This trial will be conducted in the Department of Developmental Behavioral Pediatrics, the First Hospital of Jilin University, China. One hundred high-risk infants meeting the inclusion criteria will be recruited and randomized into control and treatment groups. Only participants in the treatment group will undergo the early treatment of mouse nerve growth factor via gluteus maximus injections, 20 μg per dose, once a day, 10 consecutive days per month. The treatment will last for 6 months. Both groups will receive standard early intervention therapies. Outcome measures: The primary outcome measure is the incidence of the developmental disorders cerebral palsy and non-cerebral palsy at the actual age or corrected age of 12 months. The secondary outcome measures are Gesell Developmental Schedule scores at the actual age or corrected age of 12 months, Gross Motor Function Measure score at 6 and 12 months of treatment, and adverse events during the trial. Discussion: If treatment with mouse nerve growth factor is found to be safe and effective for the high-risk infants, new options for the early-stage clinical treatment for such infants may be developed. Ethics and dissemination: This trial has been approved by the Ethics Committee of First Hospital of Jilin University of China [approval number: 2017 (2017-290)]. This trial was designed in August 2017. Ethics approval was done in October 2017. This trial was registered in November 2017. The recruitment of subjects began in December 2017. Data analysis will be finished in December 2021. The results of the trial will be reported in a scientific conference or disseminated in a peer-reviewed journal. Anonymized trial data will be available indefinitely at www.figshare.com . Trial registration: This trial has been registered in the Chinese Clinical Trial Registry (registration number: ChiCTR-IPR-17012774). Protocol version (2.0).

The neurodevelopmental disorder Williams-Beuren syndrome is caused by spontaneous approximately 1.5 Mb deletions comprising 25 genes on human chromosome 7q11.23. To functionally dissect the deletion and identify dosage-sensitive genes, we created two half-deletions of the conserved syntenic region on mouse chromosome 5G2. Proximal deletion (PD) mice lack Gtf2i to Limk1, distal deletion (DD) mice lack Limk1 to Fkbp6, and the double heterozygotes (D/P) model the complete human deletion. Gene transcript levels in brain are generally consistent with gene dosage. Increased sociability and acoustic startle response are associated with PD, and cognitive defects with DD. Both PD and D/P males are growth-retarded, while skulls are shortened and brains are smaller in DD and D/P. Lateral ventricle (LV) volumes are reduced, and neuronal cell density in the somatosensory cortex is increased, in PD and D/P. Motor skills are most impaired in D/P. Together, these partial deletion mice replicate crucial aspects of the human disorder and serve to identify genes and gene networks contributing to the neural substrates of complex behaviours and behavioural disorders.

Background: Cerebral palsy (CP) is one of the major diseases that lead to severe disability and seriously impacts the quality of life of children. Infra-low frequency transcranial magnetic stimulation (ILF-TMS) is a new technique of noninvasive brain stimulation that exactly regulates the power of specific neurotransmitters through a special magnetic field. Our study was in order to investigate the efficacy of ILF-TMS treatment in children with spastic cerebral palsy. Methods: 113 spastic cerebral palsic children were randomly divided into two groups: conventional rehabilitation group and ILF-TMS treatment group, Healthy control group was established at the same time. In conventional rehabilitaion group, children were treated with conventional rehabilitation treatment; In ILF-TMS treatment group were treated with ILF-TMS in addition to conventional rehabilitation treatment. Neurotransmitter in the brain was recorded with encephalofluctuograph (EFG) before and after ILF-TMS treatment. Gross Motor Function Measure (GMFM), Fine Motor Function Measure (FMFM) and Gesell development scale (GDS) were used to comprehensively evaluate the motor function in children with spastic cerebral palsy. Results: The results showed that the relative power of γ-aminobutyric acid (GABA) in spastic cerebral palsy was lower than that in healthy controls and was increased significantly after ILF-TMS treatment for 3 months. The relative power of glutamate (Glu) in spastic cerebral palsy was higher than that in healthy controls and was reduced significantly after ILF-TMS treatment for 3 months. After 3 months training period there was significant improvements on the GMFM (dimension B, dimension C and dimension D), FMFM (dimension A and dimension B) and GDS (gross motor DA and gross motor DQ) in the ILF-TMS treatment group when compared to conventional rehabilitation group. Conclusions: These findings indicate that GMFM is a sensitive indicator to assess the treatment efficacy in children with spastic cerebral palsy and ILF-TMS treatment can improve the motor function through regulating neurotransmitters in brain.

Microarray technology was employed to determine the differential pattern of gene expression within the hippocampus as a result of traumatic brain injury (TBI). The validity of the microarray data was confirmed using real-time RT-PCR. Following either moderate or severe lateral fluid percussion injury, rats were studied 0.5, 4, and 24 h after injury. In general, animals exhibited mRNA up or down regulation of approximately 10% of the genes studied. However, it was clear that the pattern of gene expression was influenced by both the severity of injury and the time after injury at which animals were studied. For example, genes encoding molecules for cellular signaling, synaptic plasticity, metabolism, ion channels and transporters were up regulated following severe injury, but down regulated following moderate injury. Furthermore, moderate injury was associated with an increasing number of responsive genes as a function of time post-injury. However, animals sustaining a severe level of injury exhibited decreasing number of responsive genes during the same post-injury period. The different patterns of gene expression between injury severity and across time after the insult suggests that the pathophysiological cascade induced by TBI is accompanied by a molecular response which, like the other aspects of the cellular response for survival, may indicate a "molecular window" that may offer an opportunity for therapeutic interventions involving gene therapy. Our results also suggest that fundamentally different pathophysiological processes or cascades may be induced by different severities of injury.

BACKGROUND: Tyrosine kinase inhibitors (TKIs) have demonstrated success in the treatment of acute lymphoblastic leukemia (ALL) in patients that express BCR-ABL rearrangements (Philadelphia chromosome [Ph]). The current study aimed to assess the efficacy of TKIs and prognostic factors in the treatment of adults with Ph+-ALL. METHODS: In this multicenter retrospective study, the relationship between Ph+-ALL and treatment outcomes among Chinese patients receiving TKI-containing induction/consolidation chemotherapy was examined. A total of 86 Ph+-ALL patients were included and followed for 3.85 (0.43-9.30) years. Overall survival (OS) and event-free survival (EFS) were analyzed. RESULTS: A total of 86 Ph+-ALL patients (40 females and 46 males; median age: 34.0 years) were enrolled, including those with BCR/ABL transcripts 190 (n = 52), 210 (n = 25), and 230 (n = 2); BCR/ABL isoform determination was not available for 7 patients. Mortality was influenced by variable BCR/ABL transcripts and TKI administration, and BCR/ABL transcripts, hematopoietic stem cell transplantation (HSCT), and TKI administration were associated with the occurrence of events. The OS rate in the TKI administration group during steady state was significantly higher compared with those patients who did not receive TKI administration (P = 0.008), the EFS rate in the TKI administration group during steady state was significantly higher compared with those patients who did not receive TKIs (P = 0.012), and also higher than those with TKI salvage administration (P = 0.004). BCR/ABL transcripts 210 showed preferable OS and EFS compared with BCR/ABL transcripts 190 and 230 (P

The Sanfilippo syndrome type B is an autosomal recessive disorder caused by mutation in the gene ( NAGLU ) encoding α- N -acetylglucosaminidase, a lysosomal enzyme required for the stepwise degradation of heparan sulfate. The most serious manifestations are profound mental retardation, intractable behavior problems, and death in the second decade. To generate a model for studies of pathophysiology and of potential therapy, we disrupted exon 6 of Naglu , the homologous mouse gene. Naglu−/− mice were healthy and fertile while young and could survive for 8–12 mo. They were totally deficient in α- N -acetylglucosaminidase and had massive accumulation of heparan sulfate in liver and kidney as well as secondary changes in activity of several other lysosomal enzymes in liver and brain and elevation of gangliosides G M2 and G M3 in brain. Vacuolation was seen in many cells, including macrophages, epithelial cells, and neurons, and became more prominent with age. Although most vacuoles contained finely granular material characteristic of glycosaminoglycan accumulation, large pleiomorphic inclusions were seen in some neurons and pericytes in the brain. Abnormal hypoactive behavior was manifested by 4.5-mo-old Naglu−/− mice in an open field test; the hyperactivity that is characteristic of affected children was not observed even in younger mice. In a Pavlovian fear conditioning test, the 4.5-mo-old mutant mice showed normal response to context, indicating intact hippocampal-dependent learning, but reduced response to a conditioning tone, perhaps attributable to hearing impairment. The phenotype of the α- N -acetylglucosaminidase-deficient mice is sufficiently similar to that of patients with the Sanfilippo syndrome type B to make these mice a good model for study of pathophysiology and for development of therapy.

Sanfilippo syndrome type B (mucopolysaccharidosis III B) is a rare autosomal recessive disease caused by deficiency of alpha-N-acetylglucosaminidase, one of the enzymes required for the lysosomal degradation of heparan sulfate. The gene for this enzyme, NAGLU, recently was isolated, and several mutations were characterized. We have identified, in amplified exons from nine fibroblast cell lines derived from Sanfilippo syndrome type B patients, 10 additional mutations: Y92H, P115S, Y140C, E153K, R203X, 650insC, 901delAA, P358L, A664V, and L682R. Four of these mutations were found in homozygosity, and only two were seen in more than one cell line. Thus, Sanfilippo syndrome type B shows extensive molecular heterogeneity. Stable transfection of Chinese hamster ovary cells, by cDNA mutagenized to correspond to the NAGLU missense mutations, did not yield active enzyme, demonstrating the deleterious nature of the mutations. Nine of the 10 amino acid substitutions identified to date are clustered near the amino or the carboxyl end of alpha-N-acetylglucosaminidase, suggesting a role for these regions in the transport or function of the enzyme.

Five plastocyanin-deficient mutants were identified from a population of UV-mutagenized Chlamydomonas reinhardtii cells. Genetic complementation experiments indicated that four mutants represented alleles at the PCY1 locus (pcy1-2, pcy1-3, pcy1-4, and pcy1-5). Sequence analysis confirmed that two strains, pcy1-2 and pcy1-3, carry a frameshift (-1) and a nonsense mutation, respectively, while strains pcy1-4 and pcy1-5 synthesize an extended protein as a result of read-through mutations at the stop codon. The C-terminal extension does not affect synthesis or processing of the pre-proteins, but the polypeptides are rapidly degraded after the second (lumenal) processing event. The frameshift mutation in pcy1-2 results in loss of Pcy1 mRNA, as noted previously for strain ac208 (pcy1-1), but the abundance of Pcy1 mRNA in strain pcy1-3, which carries a nonsense mutation at codon 26, is unaffected relative to wild-type cells. The decreased abundance of frameshifted Pcy1 mRNA is attributed to increased degradation rather than decreased synthesis, since the mRNAs can be stabilized by treatment of cells with cycloheximide or anisomycin. The fifth strain has a wild-type plastocyanin-encoding gene, but the strain accumulates apoplastocyanin at the expense of holoplastocyanin. We suggest that the mutation identifies a new locus (PCY2) whose function is required for normal holoplastocyanin accumulation. Like ac208 (pcy1-1), several of the new mutants were suppressed spontaneously owing to accumulation of cytochrome c6 (a functional substitute for plastocyanin). The suppressor mutation(s) displayed Mendelian inheritance and segregated independently from the PCY1 locus, which confirms that regulation of Cyc6 expression is not tightly linked to plastocyanin function.