1.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
2.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
3.Transarterial chemoembolization combined with immune checkpoint inhibitor and molecular targeted therapy for Child-Pugh grade B hepatocellular carcinoma
Li CHEN ; Daguang WU ; Guangyu ZHU ; Binyan ZHONG ; Jinhe GUO
Journal of Interventional Radiology 2024;33(9):968-973
Objective To explore the clinical efficacy and safety of transarterial chemoembolization(TACE)combined with immune checkpoint inhibitor(ICI)and molecular targeted therapy for Child-Pugh grade B hepatocellular carcinoma(HCC).Methods The patients with Child-Pugh grade B HCC,who received TACE combined with ICI and molecular targeted therapy(combination group)or TACE monotherapy(monotherapy group)at the three medical centers including the Affiliated Zhongda Hospital of Southeast University of China between January 2018 and May 2021,were enrolled in this study.The primary outcome was overall survival(OS),and the secondary outcomes included progression-free survival(PFS),objective response rate(ORR),and clinical safety.Results A total of 126 patients were enrolled in this study,including 64 patients in the combination group and 62 patients in the monotherapy group.No statistically significant difference in median OS existed between the combination group and the monotherapy group[17.7 months(95%CI:11.9-29.9 months)vs.13.2 months(95%CI:7.8-19.9 months);P=0.160].In the combination group,the patients having a Child-Pugh score of 7 points obtained a significantly better OS[19.0months(95%CI:13.6-NR)vs.13.2 months(95%CI:8.0-NR),P=0.024].The differences in the median PFS and ORR between the two groups were not statistically significant(P=0.720 and P=0.960 respectively).Grade Ⅲ/Ⅳ adverse events occurred in 19 patients(14.1%)of the combination group and in 6 patients(9.7%)of the monotherapy group.Conclusion In treating patients with Child-Pugh grade B HCC,TACE combined with ICI and molecular targeted therapy does not show a better prognosis than TACE monotherapy,however,the patients having a Child-Pugh score of 7 points in the combination group can have a much better OS.
4.Role of different melanocyte populations in the reconstitution of pigmented hair follicles
Fang LI ; Wei CHEN ; Feiran HUANG ; Xiaoyu ZHANG ; Yuan ZHOU ; Zhenru LIU ; Daguang WANG
Chinese Journal of Dermatology 2023;56(2):118-124
Objective:To observe whether hair follicle cells from mice of different species can integrate to generate new pigmented hair follicles, and to explore the role of different melanocyte populations in pigmented hair follicle reconstruction in mice.Methods:The epidermal cell population, hair follicle epithelial cell population and dermal cell population were isolated from the skin of fetal or neonatal C57BL/6J and BALB/C mice, and epidermal melanocytes were obtained by culture and purification of the epidermal cell population. The experiments were divided into 3 parts: (1) hair follicle reconstruction experiment in neonatal C57BL/6J mice, which included 2 groups: epidermal cells + hair follicle epithelial cells group and dermal cells group; (2) chimeric hair follicle reconstruction experiment, which included 4 groups: dermal cells of neonatal C57BL/6J mice group, dermal cells of neonatal BALB/C mice group, dermal cells of neonatal BALB/C mice + dermal cells of neonatal C57BL/6J mice group, and dermal cells of fetal BALB/C mice + dermal cells of fetal C57BL/6J mice group; (3) pigmented hair follicle reconstruction experiment, which included 3 groups: dermal cells of neonatal BALB/C mice + epidermal cells of neonatal C57BL/6J mice group, dermal cells of neonatal BALB/C mice + hair follicle epithelial cells of neonatal C57BL/6J mice group, and dermal cells of neonatal BALB/C mice + cultured C57BL/6J epidermal melanocytes group. Different cells were implanted into dorsal skin fold chambers of the nude mice, and there were 4 mice in each group. At weeks 4 and 8 after inoculation, hair follicle reconstruction was assessed by gross observation, histological examination and immunofluorescence assay.Results:Among the 8 BALB/C nude mice in the 2 groups in the hair follicle reconstruction experiment, 7 survived and 1 died of wound infections on week 4 after inoculation; at weeks 4 and 8 after inoculation, no hair growth was observed in the epidermal cells + hair follicle epithelial cells group (3 mice) , while normal hair grew out in the dermal cells group (4 mice) mixed with epithelial components. Among the 16 BALB/C nude mice in the 4 groups in the chimeric hair follicle reconstruction experiment, 14 survived and 2 died of wound infections on week 4 after inoculation; at weeks 4 and 8 after inoculation, brown-grey hair grew well in the dermal cells of neonatal BALB/C mice + dermal cells of neonatal C57BL/6J mice group (4 mice) , and dermal cells of fetal BALB/C mice + dermal cells of fetal C57BL/6J mice group (3 mice) . Among the 12 BALB/C nude mice in the 3 groups in the pigmented hair follicle reconstruction experiment, 10 survived and 2 died of wound infections on week 4 after inoculation; at weeks 4 and 8 after inoculation, only white hair grew out in the dermal cells of neonatal BALB/C mice + cultured C57BL/6J epidermal melanocytes group (3 mice) , and no hair follicle melanocytes were observed by immunofluorescence assay, while brown-grey hair grew well in the dermal cells of neonatal BALB/C mice + epidermal cells of neonatal C57BL/6J mice group (4 mice) , and dermal cells of neonatal BALB/C mice + hair follicle epithelial cells of neonatal C57BL/6J mice group (3 mice) .Conclusions:The interaction between mesenchymal cells and hair follicle epithelial cells is a necessary condition for hair follicle reconstruction. The hair follicle cells from different species of mice can integrate to generate new pigmented hair follicles. Both hair follicle melanocytes and epidermal melanocytes can participate in the formation of pigmented hair follicles, but differentiated melanocytes have no such ability.
5.Treatment of pincer nails
Yuan ZHOU ; Wei CHEN ; Daguang WANG
Chinese Journal of Dermatology 2019;52(5):364-367
Pincer nails are a kind of nail deformity characterized by abnormal thickening and distal transverse overcurvature of the nail plate,whose etiology and pathogenesis are still unclear.According to the morphology of the nail plate,pincer nails can be divided into three subtypes:common pincer nail (trumpet nail),tile-shaped nail,and plicated nail.Width index,height index and curvature index are objective indicators to evaluate the severity and therapeutic effect of the pincer nails.According to the classification and severity of the pincer nails,appropriate treatment methods can be adopted to improve the appearance of the nail plate and relieve clinical symptoms of patients.This review systematically summarizes the classification of,evaluation of the severity of and several therapeutic methods for pincer nails.
6.Effects of recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immta ure dendritic cells on T cell proliferation
Zhe CHEN ; Mingdao HU ; Daguang TIAN ; Xiaoping WEI ; Henghai YU ; Peng CHEN
Journal of Medical Postgraduates 2017;30(1):5-9
Objective HLA-G widely participates in immune tolerance by its combination with immunoglobulin-like tran-scripts IL-2 and IL-4 on the surface of dendritic cells (DCs).The aim of the article was to explore the effects of recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells on T cell proliferation . Methods Marrow blood was collected from macaca mulattas by the puncture needle after anesthesia .Density gradient centrifugation method was applied in separating mononuclear from the extracted blood on which CD 34+cells were collected and pu-rified by means of immunomagnetic separation .Small doses of cyto-kines were added to get the immature dendritic cells after induced dif-ferentiation of CD34+cells.After the recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells , observation was done on the viral infection efficiency and western blot was used in detecting the expression of HLA -G in immature den-dritic cells.Taking T cells in macaca mulatta as responders and DCs transfected by recombinant adnovirus -mediated HLA-G as stimu-lators, mixed lymphocyte test was conducted .T cells were divided into 5 groups: mDC group ( mature DCs ) , imDC group ( immature DCs), imDC(L) group(addition of 100 ng/mL lipopolysaccharide after getting imDC at 7th day) , imDC(V) group (imDCs infected by recombinant adnovirus-mediated HLA-G) , imDC( L+V) group ( imDCs infected by recombinant adnovirus-mediated HLA-G along with the addition of 100 ng/mL lipopolysaccharide in culture process ) . Results We obtained the immature dendritic cells and recom-binant adenovirus of HLA-G expressed in these cells .Flow cytometry showed DC purity was up to 92.3 %, imDC purity was up to 72.39%and positive percentage of CD 4+T was greater than 80%.In comparison with imDC group ,the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified (P<0.01).In comparison with imDC(V) group, the proliferation of stim-ulated T cells in imDC, mDC, imDC(L), and imDC(L+V) groups was obviously intensified (P <0.01).In comparison with imDC(L+V) group, the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified(P<0.01). Conclu sion Im-mature DCs infected by recombinant adnovirus can inhibit the proliferation of T cells effectively .
7.Study of introperitoneal hyperthermic perfusion chemotherapy combined with systemic neoadjuvent chemotherapy in treatment of gastric cancer patients with peritoneal carcinomatosis.
Daguang WANG ; Yanpeng XING ; YuChen GUO ; Yang ZHANG ; Yujia CHEN ; Jian SUO
Chinese Journal of Gastrointestinal Surgery 2016;19(5):540-544
OBJECTIVEThe aim of this study is to discuss the curative effect of introperitoneal hyperthermic perfusion chemotherapy(IHPC) combined with systemic neoadjuvant chemotherapy on the gastric cancer patients with peritoneal carcinomatosis.
METHODSSixty-four patients with gastric cancer and peritoneal carcinomatosis who were hospitalized in the Department of Gastrointestinal Surgery of First Hospital of Jilin University from December 2006 to December 2013. After peritoneal carcinomatosis was confirmed during laparoscopic exploration, FOLFOX6 (oxaliplatin and calcium folinate and 5-Fu) was performed for systemic chemotherapy. One course was 14 days and a complete treatment includes four courses. At the same time, patients underwent peritoneal catheter insertion and received IHPC(5-Fu 1 500 mg/m(2) and Cisplatin 35 mg/m(2) were added into 0.9% NaCl solution 2 000 ml, the infusion velocity was 35-45 ml/min, infusion time was 45-60 minutes, the temperature was controlled to 41°C). A comprehensive evaluation was taken after the fourth course of treatment before operation. Further surgical therapy was performed according to the assessment result.
RESULTSSixty-four patients received IHPC combined with systemic chemotherapy. Thirty-two patients(50.0%) had partial response, 18(28.1%) stable disease, and 14(21.9%) progressive disease after chemotherapy. No severe complications or death occurred during the neoadjuvant chemotherapy. Thirty-two patients(50.0%) received radical resection, 10(15.6%) palliative operation, and another 22 patients(37.4%) didn't comply with inclusion criteria of operation. Patients receiving operation had a median survival time of 678 days, which was significantly longer than patients without operation, with a median survival time of 251(χ(2)=23.34, P=0.02).
CONCLUSIONSIHPC combined with systemic chemotherapy is an effective therapeutic method for gastric cancer patients with peritoneal carcinomatosis in terms of reducing preoperative tumor load and achieving radical resection.
Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma ; drug therapy ; Chemotherapy, Cancer, Regional Perfusion ; Cisplatin ; therapeutic use ; Combined Modality Therapy ; Digestive System Surgical Procedures ; Fluorouracil ; therapeutic use ; Humans ; Hyperthermia, Induced ; Laparoscopy ; Leucovorin ; therapeutic use ; Neoadjuvant Therapy ; Organoplatinum Compounds ; therapeutic use ; Peritoneal Neoplasms ; drug therapy ; Peritoneum ; Stomach Neoplasms ; drug therapy ; Treatment Outcome
8.A cross-sectional study of parental self-reported physical violence against their children
Jingqi CHEN ; Yanqiu KONG ; Xiangmei LI ; Jingyi LI ; Yanan FENG ; Xiaoxia ZHAO ; Wenjing ZHANG ; Xiaoling LOU ; Daguang CHEN ; Yichen JIN ; Buyi YU ; Chen QIU
Chinese Mental Health Journal 2015;(9):703-707
Objective:To investigate the prevalence of physical violence against children (PVAC)by parents and to explore possible related factors.Methods:In two primary schools in Liaoning Province,1164 parents of pupils in grade 1 to 6 were surveyed by a self-administered questionnaire anonymously to analyze situation about PVAC and related factors.PVAC was defined as having one or more following parent-to-child behaviors in the past three months:push,or shook a child;pinch,screw,or scratch a child;hit child's buttocks with hand;hit child's hand,foot,arm,leg or back with hand;hit child's buttocks with an object;hit child's face or head with hand;hit elsewhere (not buttocks)with an object;kicked a child with a foot or hit with a fist.Results:Of 1164 parents,53.1% reported that they had minor PVAC (51.0%)or/and severe PVAC (19.8%).Multivariate logistic regres-sion analysis indicated that following factors increased the risk of PVAC:child's male gender (OR =1.49),younger age of children (OR =1.38),poor school performance of children (OR =1.85 ),mother (OR =2.09),parents'childhood physical violence victimization experiences (OR =1.53),parents'supportive or tolerant attitudes towards corporal punishment (OR =3.15),parents'lower awareness of the harmfulness of physical violence against children (OR =2.31),and lower social economic status (OR =1.47).Conclusion:The PVAC by their parents may be com-mon in the present study.Parents'supportive or tolerant attitudes toward corporal punishment and low awareness of the harmfulness of physical violence is the main risk factors of PVAC.
9.Techniques and feasibility of laparoscopic total gastrectomy combined with distal pancreatectomy and splenectomy
Daguang WANG ; Yang ZIANG ; Xuan SUN ; Jinhai YU ; Yan CHEN ; Liang HE ; Huai CHEN ; Jian SUO
Chinese Journal of Digestive Surgery 2012;11(2):132-135
Laparoscopic curative gastrectomy has been widely adopted because it is minimal invasiveness.The efficacy of laparoscopic gastrectomy is comparable to that of open gastrectomy for patients with early or advanced gastric cancer,but few studies of laparoscopic total gastrectomy combined with distal pancreatectomy and splenectomy for patients with advanced gastric cancer have been reported.Seven patients with advanced gastric cancer received laparoscopic total gastrectomy combined with distal pancreatectomy and splenectomy at the First Hospital of Jilin University from September 2010 to September 2011.The mean operation time,blood loss,postoperative hospital stay and the nunber of lymph nodes resected were (218 ± 24) minutes,(366 ± 174) ml,( 14.6 ± 2.8 ) days and 32 ± 15,respectively.Two patients were complicated by pancreatic juice leakage and were cured with drainage. No anastomotic leakage,abdominal bleeding or mortality was observed. With the development of techniques and equipments,laparoscopic total gastrectony combined with distal pancreatectomy and splenectomy is safe and feasible in selected patients.
10.Therapeutic effect assessment of integrated intervention for learning difficulties in children
Guixiu LIN ; Shan CHEN ; Daguang CHEN ; Jun HU ; Xiaorong YIN
Chinese Journal of Tissue Engineering Research 2005;9(24):210-212
BACKGROUND:Children with learning difficulty often present with cognitive impairments and imbalanced development of intelligence accompanied by emotional and behavioral problems.OBJECTIVE:To evaluate the therapeutic effects of integrated interventions in multiple aspects for children with learning difficulties.DESIGN:A case-control study of children with learning difficulties.SETTING:Department of Pediatric Internal Medicine, Union Hospital Affiliated to Fujian Medical University.PARTICIPANTS:This study involved school-age children of 6 to 12years old with learning difficulties who came to the department specialized in pediatric psychology for consultation from July 1998 to July 2001. The diagnosis of learning difficulties conformed to the diagnostic criteria for special learning disability in ICD-10 with IQ > 70 as an essential condition. Children with emotional disorders, mental deficiency, hyperactivity,visual and auditory dysfunctions and organic cerebral diseases were excluded. The diagnosis was further verified by a professor specialized in neuropsychology. The children voluntarily participated in the cognitive training, behavioral intervention and comprehensive trainings of senses and those who fulfilled the one-year treatment were included in this study. The ratio of therapists to the children was 1:2, and each session of treatment lasted for 90-120 minutes given 2-3 times a week, and a treatment course consisted of 24 sessions. The children were supposed to complete 4 consecutive treatment courses. Totally 31 children completed the treatment courses, including 24 boys and 7 girls with the mean age of(7.7±1.2)years who had an average IQ of 87.5±8.6.grated interventions for one year consisting of cognitive training, behavioral intervention and comprehensive training of the senses. Raven's Deductive Reasoning test and Beery Visual-Motor Integration Test were use to examine the deductive reasoning ability and visual-motor integration of the children, and visual attention test and auditory attention test, as well as sequential and reverse-sequence memory tests were employed to test their attentions. Achenbach children behavioral scale was applied to evaluate the children's behavioral problems, and Conners' questionnaire for children's behavioral problem was used to measure their attention-deficit/hyperactivity disorder (ADHD) index.MAIN OUTCOME MEASURES:Changes in the deductive reasoning ability, capability of visual-motor integration, attention and psychological behaviors after one year of intervention.RESULTS:All the 31 patients children entered the result analysis with no loss. After one year of integrated intervention, the children had significantly improved scores for Raven's Deductive Reasoning Test (87.51 ±8.64 vs 90.03±8.70, t=-9.610, P < 0.001) and Beery Visual-Motor Integration Test (6.75±1.74 vs 8.27±1.94, t=-9.002, P < 0.001). Obvious improvement was also seen in the visual attention test (15.90±20.11 vs 2.74±3.67), scope of auditory attention and memory (7.70±1.60 vs 11.23±1.23for sequential memory and 3.52±0.93 vs 6.97±0.16 for reversed-sequence memory, P < 0.01). The total scores for ADHD assessment declined to different extents at 3 and 6 months during the treatment and upon the completion of the treatment (22.65±3.53 vs 18.87±4.14 at month 3, 5.65±3.92at month 6 and 12.65±3.36 at one year, t=7.35-15.36, P < 0.01). The scores for 9 psychobehavioral items in Achenbach children behavioral scale obviously declined were also reduced significantly during the therapy,and 87% (26/31) of the children had obviously improved academic performance.CONCLUSION:Interventions integrating cognitive training, behavioral intervention and comprehensive training of the senses may produce good and persistent effect on the cognitive and emotional symptoms of children with learning difficulties, resulting in obviously improved visual, auditory and brain functions.

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