Objective To investigate the symptoms and affecting factors of food intolerance among 903 adults.Methods Food special antibody(IgG)in human serum were examined by ELISA.Food Intolerance Health Assessment Questionnaire was used to assess positive rate of food intolerance and food intake habits.Chi-square test and Mann-Whitney test were used for data analysis.Results The ratio of food intolerance was 0.33％ to 28.13％.Intolerant foods include crab(28.13％),yolk(27.02％),milk (13.07％),soybean(11.96％),shrimp,tomato,corn,beef,rice,chicken,mushroom,wheat,and pork.The symptoms of nervous system(55.89％),digestive system(45.69％)and respiratory system(15.64％)were commonly seen.Prevalence of food intolerance in mental labors was lower than manual labors.Conclusion The most common intolerant food includes crab,yolk and milk.The symptoms of food intolerance are different.Manual labors are more susceptible to food intolerance.

Objective To established a diagnostic methods to idenfity Niemann-Pick disease type C (NPC) in China. Methods Two patients aged 5 and 20 years respectively who presented progressive neurologic regression and splenomegaly were subjected to filipin staining of cultured skin fibroblasts and genetic analyses of NPC1 gene. Results Although there were differences in onset ages and clinical presentations, filipin staining of the cultured skin fibroblasts confirmed the clinical diagnosis of NPC, showing an intense punctate pattern of fluorescence concentrating around the nuclei, consistent with the accumulation of unesterified cholesterol in NPC cells. Genetic sequence analysis further verified the results of filipin staining. The case 1 was compound heterozygous for M1142T(3425T＞C),R1186H(3557T＞C)and case 2 for Q88H(264G＞T),469_470insGT.The latter 2 mutations were novel, and the possibility of polymorphisms was not supposed by analyzing 134 DNA samples obtained form normal controls. Conclusions Filipin stainning of the cultured skin fibroblasts is a reliable method to clinically diagnose NPC with a sensitivity. Genetic diagnose should be performed where genetic analysis is allowed.

ObjectiveTo investigate clinical symptoms and influencing factors of food intolerance and to identify susceptibility of food intolerance among adults with various Traditional Chinese Medicine (TCM) constitutions.MethodsA total of 411 individuals were recruited by using simple sampling method.ELISA was used to test serum levels of food intolerance specific IgG antibody.TCM constitution,food intake habit,lifestyle,and biochemical data were collected.Results ( 1 ) The prevalence of food intolerace was 60.1％ (247/411).(2) The most commonly seen food intolerance specific IgG were those against egg (44.9％ ),crab ( 37.7％ ),or cod ( 23.1％ ).Intolerance to 1 to 5 kinds of food could be found in single individual.Subhealthy people were more likely to suffer from food intolerance ( P＜0.05 ).( 3 ) Those with food intolerance always presented with various symptoms,mainly respiratory system symptoms.(4) The most common types of TCM constitution of the participants were gentleness constitution,qi-insufficiency constitution,yang-deficiencyconstitution,anddampness-heatconstitution.Thoseofnon-bloodstasis constitution were more likely to suffer from crab intolerance ( P＜0.05 ).Conclusion The prevalence of food intolerance is relatively high in Fuzhou areas.Subhealthy status,obesity women,and pathological constitutions may be risk factors of food intolerance.

In order to better understand the clinical manifestation of systemic lupus erythematosus (SLE) with intracranial hypertension syndrome (IHS), we analyzed the clinical features and treatment of a typical SLE patient with IHS. SLE is one of the most unpredictable autoimmune diseases involving multiple organ systems that is defined clinically and associated with antibodies directed against cell nuclei. IHS is an uncommon manifestation of neuropsychiatric SLE (NPSLE) and is characterized by an elevated intracranial pressure, papilledema, and headache with occasional abducens nerve paresis, absence of a space-occupying lesion or ventricular enlargement, and normal cerebrospinal fluid chemical and hematological constituents. IHS has been reported in a few sporadic cases in patients with SLE worldwide, but rarely has been reported in China. In this study, a 34-year-old female SLE patient with IHS was reported and pertinent literature reviewed. The clinical presentation, image logical features, and investigatory findings were discussed.
Diagnosis, Differential ; Intracranial Hypertension/diagnosis ; Intracranial Hypertension/*etiology ; Lupus Erythematosus, Systemic/*complications ; Lupus Erythematosus, Systemic/diagnosis

OBJECTIVE: To analyze ultrasonographic finding in fetuses with Wolf-Hirschhorn syndrome (WHS) and refine the critical region on chromosome 4p16.3 for WHS-associated fetal growth retardation (FGR). METHODS: In total 2262 fetuses with abnormal ultrasonographic findings who underwent prenatal karyotyping and chromosomal microarray analysis were reviewed. WHS-associated 4p deletions detected in these fetuses were compared, and prenatal ultrasound findings in such fetuses were summarized. Meanwhile, WHS cases with prenatal ultrasound findings and isolated 4p deletions in previous studies were included for further analysis. An analysis of smallest region of overlap (SRO) among discrepant 4p deletions in these cases above was performed to define a critical region for FGR. RESULTS: 4p deletions were detected in 10 of the 2262 fetuses and 5.0% of the 202 fetuses with FGR. Combined with 80 WHS cases from previous studies, the most common prenatal ultrasound finding was FGR, which yielded a frequency of 76.7%. In addition, a SRO spanning approximately 419 kb (genomic position: 1.32-1.74 Mb) on chromosome 4p16.3 was discovered by comparing the unusual 4p deletions among the 10 fetuses. The region contained seven protein-coding genes, including TACC3, SLBP, TMEM129, FAM53A, MAEA, UVSSA and CRIPAK. CONCLUSION For fetuses with WHS, the most common prenatal ultrasound phenotype was FGR. A region between 1.32 Mb to 1.74 Mb from the telomere on chromosome 4p16.3 is critical for WHS-associated FGR, for which TACC3 and SLBP are the candidate genes.

In order to better understand the clinical manifestation of systemic lupus erythematosus (SLE) with intracranial hypertension syndrome (IHS),we analyzed the clinical features and treatment of a typical SLE patient with IHS.SLE is one of the most unpredictable autoimmune diseases in-volving multiple organ systems that is defined clinically and associated with antibodies directed against cell nuclei.IHS is an uncommon manifestation of neuropsychiatric SLE (NPSLE) and is characterized by an elevated intracranial pressure,papilledema,and headache with occasional ab-ducens nerve paresis,absence of a space-occupying lesion or ventricular enlargement,and normal cerebrospinal fluid chemical and hematological constituents.IHS has been reported in a few sporadic cases in patients with SLE worldwide,but rarely has been reported in China.In this study,a 34-year-old female SLE patient with IHS was.reported and pertinent literature reviewed.The clinical presentation,image logical features,and investigatory findings were discussed.

In order to explore the role of TNF-a in Niemann-Pick type C (NPC) disease,lentivi-ral-delivered RNA interference (RNAi) was used to silence the expression of murine TNF-a gene in vitro and in npc mice.Interference efficiency of the lentivirus expressing TNF-a-siRNA,previously constructed with the concentration of 2×108ifu/mL,was determined by RT-PCR and ELISA in BV-2 tricularly infused into 4-week old npc mice for a 4-week period,and the mice were divided into 3 (n=4).By using immunohistochemistry and real-time PCR,the down-regulation of the target genes ciently in vitro and the interference efficiency was 66.7%.Lentivirus could be expressed stably for long-term in the npc mice brain.Immunohistochemistry and real-time PCR revealed that,as com-pard with non-intervention gronp and Lenti-control group,Lenti-TNF-a-siRNA efficiently down-regulated the expression of murine TNF-a gene with the interference efficiency being 66.9%.TNF-a-siRNA downregulated the expression of TNF-a gene in vitro and in vivo,which provided a potential tool for studying and treation neurodegenerative diseases and TNF-a-related diseases.

Objective:To investigate the role of the clearance of exogenous myelin antigen in experimental autoimmune encephalomyelitis (EAE).Methods:EAE was induced in C57BL/6J mice by subcutaneous immunization with myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) or FITC-MOG 35-55. The concentration of exogenous myelin antigen was assessed by analyzing the proliferation of the transferred CFSE-labeled mT/mG-2D2 CD4 + T cells in spleen tissues. The release of exogenous myelin antigen from the inoculation sites was analyzed by immunohistochemistry and flow cytometry. HE staining was used to investigate the mechanism underlying the rapid clearance of exogenous myelin antigen. The role of the clearance of exogenous myelin antigen in EAE was investigated by comparative analysis of EAE induced by subcutaneous immunization in the back and footpads, and analyzing the therapeutic effect of soluble MOG 35-55. Results:The proliferation of mT/mG-2D2 CD4 + T cells in mice was enhanced on day 2 than on day 7 after immunization [(52.6±6.8)% vs (18.5±4.9)%, P<0.01]. There was no significant difference in the proliferation of mT/mG-2D2 CD4 + T cells between EAE mice (day 13) and naive mice [(4.4±1.5)% vs (2.5±1.4)%, P=0.11]. Immunohistochemistry and flow cytometry showed that MOG 35-55 was released and engulfed by CD11b + cells at the inoculation sites, and no more MOG 35-55 was released at the onset of EAE. HE staining showed that granuloma that formed surrounding the antigen emulsion during EAE development prevented antigen release from the emulsion, completely isolating the antigen from the peripheral immune system. The incidence of EAE was relatively low in mice immunized via footpads, which was related to the sustained release of MOG 35-55, but had no direct relation to CD4 + regulatory T cells. Continuous intraperitoneal injection of soluble MOG 35-55 could prevent and treat EAE. Conclusions:Exogenous myelin antigen has been completely cleared in EAE mice, and the occurrence of EAE depends on the clearance of the myelin antigen.

Objective:To investigate the methylation status of SDC2, PPP2R5C and ADHFE1 genes in stool and their values in the screening of colorectal cancer and precancerous lesions.Methods:From August 2020 to March 2021, 64 patients with colorectal cancer, 72 patients with adenoma, 33 patients with hyperplastic polyps and 59 healthy people were recruited from Qingdao Central Hospital Affiliated to Qingdao University, and the morning stool samples were collected from the research subjects. The genomic DNA was extracted and modified with sulfite. The methylation status of SDC2, PPP2R5C and ADHFE1 genes were detected by methylation specific polymerase chain reaction (MSP), and the fecal occult blood test (FOBT) was performed. Taking the pathological results as the gold standard, receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to compare the effect of combined detection of methylation of three genes and FOBT in predicting colorectal cancer and precancerous lesions. R-Studio software was used to construct a nomogram for the prediction of colorectal cancer with combined detection of gene methylation in stool and other clinical features, and the calibration and validation were performed.Results:The positive rates of combined detection of methylation of SDC2, PPP2R5C and ADHFE1 genes in stool were higher than those of FOBT in colorectal cancer+adenoma [74.3% (101/136) vs. 47.1% (64/136), χ2 = 23.20, P = 0.001], colorectal cancer [90.6% (58/64) vs. 70.3% (45/64), χ2 = 8.91, P = 0.003] and adenoma [59.7% (43/72) vs. 26.4% (19/72), χ2 = 14.43, P = 0.002]. There was no significant difference in the positive rates in hyperplastic polyps [21.2% (7/33) vs. 6.1% (2/33), χ2 = 0.12, P = 0.125] and healthy controls [10.2% (6/59) vs. 8.5% (5/59), χ2 = 4.01, P = 1.000]. The combined detection of gene methylation was better than FOBT in the prediction of colorectal cancer + adenoma [AUC: 0.85 (95% CI 0.80-0.91) vs. 0.71 (95% CI 0.64-0.78), P < 0.05], especially in the prediction of adenoma [AUC: 0.82 (95% CI 0.74-0.89) vs 0.64 (95% CI 0.57-0.69), P < 0.001]. The sensitivity and specificity of ADHFE1 gene methylation status in predicting colorectal cancer were high (90.6% and 96.6%). In colorectal cancer patients over 50 years old, the positive rate of combined detection of gene methylation was higher than that of FOBT [90.2% (55/61) vs. 68.9% (42/61), P < 0.05]. The nomogram calibration curve for predicting colorectal cancer constructed based on the combined detection of gene methylation and each clinical feature showed a high degree of concordance between the predicted and observed diagnostic performance of colorectal cancer. Conclusions:The methylation levels of SDC2, PPP2R5C AND ADHFE1 genes in stool are increased in patients with colorectal cancer or adenoma. The combined detection of gene methylation is expected to be a non-invasive method for the screening of colorectal cancer and precancerous lesions.

ObjectiveTo study the effect of Bushen Huoxuetang on the apoptosis and the expression of B-cell lymphoma （Bcl-2）-associated X protein （Bax）/ Bcl-2 and cleaved cysteine-containing aspartate proteolytic enzyme-3 （cleaved Caspase-3） in the nude mouse model of bone metastasis of breast cancer， and explore the mechanism of Bushen Huoxuetang in inhibiting bone destruction. MethodThirty BALB/c female nude mice were randomly assigned into blank group （n=6） and model group （n=24）. The suspension of 4T1 breast cancer cells was injected into the tibia of mouse right lower limb to establish model of bone metastasis of breast cancer. The successfully modeled nude mice were randomly assigned into model group， Bushen Huoxuetang group， zoledronic acid group， and combined drug group， with 6 mice in each group. Bushen Huoxuetang was administrated at a dose of 36.67 g·kg^-1， once a day， and zoledronic acid was administrated by subcutaneous injection at a dose of 100 μg·kg^-1， twice a week. The combined drug group was administrated with the same doses of Bushen Huoxuetang group by gavage and zoledronic acid by subcutaneous injection. The mice in the blank group and the model group were administrated with the same volume of distilled water by gavage for 14 days. On the next day at the end of drug administration， the mice were sacrificed by cervical dislocation. The general situation and weight changes of the mice were examined. The right lower limb was collected， and X-ray scanning and hematoxylin-eosin （HE） staining methods were used for observation of pathological changes in the bone. The terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） was employed to detect the apoptosis of bone tissue in nude mice， and Western blot to determine the expression of Bax/Bcl-2 and cleaved Caspase-3 in the bone tissue. ResultCompared with the blank group， the modeling reduced the body weight （P<0.01） and increased the right lower limb weight of the nude mice （P<0.01）. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination increased the body weight （P<0.01） and decreased the right lower limb weight （P<0.01）. Compared with the blank group， the other groups showed obvious tumor cell atypia， deep nuclear staining， and clear bone metastasis， and the model group showed obvious osteolytic damage in right lower limb and loss of proximal tibia and knee joint. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination reduced the osteolytic lesions in the right lower limb and recovered part of the bone structure， demonstrating an inhibitory effect on bone destruction. The TUNEL assay showed that the model group had lower apoptosis rate of bone metastatic tumor cells than the blank group， Bushen Huoxuetang group， zoledronic acid group， and combined drug group （P<0.01）. Compared with the blank group， the modeling down-regulated the expression of Bax and cleaved Caspase-3 （P<0.01） and up-regulated the expression of Bcl-2 （P<0.01）. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination up-regulated the expression of Bax （P<0.01） and cleaved Caspase-3 （P<0.05， P<0.01） and down-regulated the expression of Bcl-2 （P<0.05， P<0.01）. ConclusionBushen Huoxuetang may inhibit bone destruction in the nude mouse model of bone metastasis of breast cancer by up-regulating the expression of Bax， down-regulating the expression of Bcl-2， activating cleaved Caspase-3， and further inducing apoptosis.