T),one dinucleotide deletion(1801-02 del AG) and one base insertion(1125 ins GCCATCG),which resulted in eight missense mutations,two nonsense mutations and two frame shift mutations,namely P534R,G343V,R259W,A141T,R401Q,K276E,Y174C,A314P,S108X,Q177X,fs E471 and fs A247.Conclusion The combined DHPLC and sequencing approach may act as a rapid and efficient method for ABCD1 gene mutation analysis in patients and carriers of X-linked adrenoleukodystrophy families.There exist different ABCD1 gene mutations in different pedigrees,and no obvious correlation between the genotype and phenotype has been found.

Objective Membranous nephropathy ( MN) is rarely complicated by crescents.This study was to observe the clinicopathologic characteristics of MN with crescents. Methods This retrospective study included 53 cases of biopsy-proven idiopathic MN with crescents in the absence of immunologic and clinical etiologic factors and another 100 MN patients without histological crescents as controls.The clinicopathologic features, treatment response, and out-comes were analyzed and compared between the two groups of pa-tients. Results Significantly higher percentages of hypertension and decreased eGFR were observed in the MN +crescents group than in the control (47.2%vs 19.0%, P<0.05;28.3%vs 40.%, P<0.05).Circulating autoantibodies against the M-type phospholipase A2 receptor (PLA2R) were found in 66.7%(30/45) of the patients.The glomeruli exhibited a median of 4.6%(1.8%-35.3%) involvement of crescents.Compared with the controls, the MN +crescents group showed remarkably higher rates of segmental glomer-ulosclerosis lesions (16.0%vs 49.1%, P<0.05), capillary loops necrosis (0.0%vs 11.3%, P<0.05), interstitial fibrosis/tubu-lar atrophy (IFTA) (54.0%vs 86.8%, P<0.05) and afferent arterial lesions (65.0%vs 92.5%, P<0.05).No significant differ-ences were found in the outcomes of the two groups of patients. Conclusion MN with crescents is rare, and secondary MN and cres-centic glomerulonephritis should be considered.Crescentic MN usually presents with hypertension and renal dysfunction clinically and high rates of severe segmental and global glomerulosclerosis, capillary loops necrosis, and IFTA histologically.The condition has a fa-vorable short-term prognosis.

Objective To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy(X-ALD).Methods The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells.Maternal contamination was evaluated by paternity test.PCRRFLP,sequencing and denaturing high performance liquid chromatography(DHPLC)were used to detect the ABCD1 gene of fetal genome.Results In the pedigree 1,the PCR product(799 bp)of the fetus 1 and her father(normal control)could be digested with BcnI. No P560L mutation,which was present in the index patient,was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing.In the pedigree 2,the PCR product(232 bp)of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation,which was present in the propositus,was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing.In the pedigree 3,the PCR product(271 bp)was digested with AciI.the pattern of the fetus 3 and the propositus being the same,and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing.In the pedigree 4,the PCR product(269 bp)was analyzed with the DHPLC,and the pattern of elution peaks of the fetus 4 and her father was similar,but different from that of the propositus.No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing.Judging from the sex of the fetuses,fetuses 1 and 2 were normal homozygotes,fetus 3 was an ALD hemizygote,and fetus 4 was a normal hemizygote.Conclusion A new protocol for X-ALD prenatal molecular diagnosis is proposed,which would ensure the accuracy of prenatal diagnosis.

OBJECTIVETo verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 (CASK/Id1) pathway in fibroblasts of human keloid.

METHODSImmunofluorescence laser was used to confirm CASK and Id1 protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Id1 expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Id1 protein of keloid fibroblasts was tested by immunoprecipitation.

RESULTSCASK and Id1 protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Id1 were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0.658 +/- 0.024, which was lower than that in the normal control group (1.076 +/- 0.008, t = 11.159, P < 0.05). The expression of Id1 mRNA was 0.497 +/- 0.014, which was higher than that in the normal control group (0.307 +/- 0.017, t = 15.148, P < 0.05). The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0.057 +/- 0.006, which was lower than that in the normal control group (0.168 +/- 0.012, t = 13.524, P < 0.05); the expression of Id1 protein was 0.812 +/- 0.035, which was higher than that in the normal control group (0.368 +/- 0.031, t = 16.356, P < 0.05). The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Id1 precipitate. There was a natural binding of CASK and Id1 in keloid fibroblasts.

CONCLUSIONCASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

Cell Proliferation ; genetics ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; metabolism ; Fibroblasts ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction

Objective To investigate the effects of liraglutide on glucose-lipid metabolism in ApoE-/-mice with RNAi-mediated adiponectin gene inhibition. Methods The dose-effective relationship of liraglutide was evaluated by intravenous glucose tolerance test (IVGTT), and the insulin sensitivity and glucose-lipid metabolism were assessed by the hyperinsulinemic-euglycemic clamp technique using 3-[3 H]-glucose as a tracer. Results In the IVGTT, blood glucose was significantly lower in the 1 mg/kg liraglutide group than that in other groups ( all P＜0. 01 ) at the points of 5, 15, and 30 min after glucose load. However, plasma insulin was significantly higher at the points of 5 and 15 min (all P＜0. 01 ). Fasting blood glucose (FBG), body weight, free fatty acids (FFA),total cholesterol, triglycerides, low-density lipoprotein-cholesterol ( LDL-C), and fasting plasma insulin in ApoE-/-mice with co-injection of liraglutide and adiponectin shRNA adenovirus ( HEA group ) were significantly lower than those in ApoE-/-mice with adiponectin shRNA adenovirus injection ( ADI group, P＜0. 05 or P＜0. 01 ). However,high-density lipoprotein-cholesterol (HDL-C) was significantly higher than the latter (P＜0. 05 ). During the steady-state of clamp, plasma insulin in ADI group was significantly higher than that in HEA group (P＜0. 01 ). Although FFA, total cholesterol, and triglycerides were suppressed in all groups, they were still higher in ADI group than those in HEA group (P＜0. 05). Glucose infusion rate (GIR) in HEA group were significantly higher than that in ADI group ( P ＜ 0. 01 ). At the end of clamp, glucose disappearance rate ( GRd ) was significantly lower, and hepatic glucose production significantly higher in ADI group than those in HEA group (P＜0.01 ). Conclusion Administration of liraglutide may ameliorate insulin resistance via increasing plasma adiponectin level in ApoE-/-mice with RNAi-mediated adiponectin gene inhibition.

Objective:To investigate the differences in transcranial Doppler (TCD) hemodynamic parameters and their correlation with cognitive impairment in middle-aged and elderly populations with different cognitive states in rural communities.Methods:From January to February 2022, participants were recruited from rural communities in Yandu District, Yancheng for cognitive assessment and TCD examination. TCD parameters were recorded, including mean flow velocity (MFV), peak systolic velocity (PSV), pulsatile index, and resistance index of bilateral anterior cerebral artery (ACA), middle cerebral artery (MCA), posterior cerebral artery (PCA), vertebral artery and basilar artery. According to the Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) scores, the participants were divided into a normal cognitive function group, a mild cognitive impairment (MCI) group, and a dementia group. Spearman correlation analysis was used to investigate the correlation between hemodynamic parameters and blood pressure. Multivariate logistic regression analysis was used to identify the independent influencing factors of MCI and dementia, and multiple linear regression analysis was used to identify the correlation between hemodynamic parameters and cognitive assessment scale scores. Results:A total of 270 participants were enrolled, including 82 in the normal cognitive function group, 100 in the MCI group, and 88 in the dementia group. The comparison between groups showed that advanced age, female and low education years might be the risk factors for dementia in rural areas. The proportion of smoking and drinking in the dementia group was lower than that in the normal cognitive function group and the MCI group. The right ACA MFV and PSV in the dementia group were significantly lower than those in the normal cognitive function group and the MCI group, while the left PCA PSV was significantly lower than that in the MCI group (all P<0.05). Multivariate logistic regression analysis showed that compared with the normal cognitive function group, the right ACA PSV (odds ratio [ OR] 0.964, 95% confidence interval [ CI] 0.935-0.993; P=0.016) was an independent related factor associated with dementia; compared with the MCI group, right ACA PSV ( OR 0.967, 95% CI 0.936-0.999; P=0.043) and left PCA PSV ( OR 0.953, 95% CI 0.911-0.998; P=0.039) were the independent related factors for dementia. Multiple linear regression analysis showed that the right ACA PSV ( β=0.431, P=0.036) and left PCA PSV ( β=0.133, P=0.021) were the independent related factors for MMSE score. Spearman correlation analysis showed that there was a significant correlation between blood pressure and several hemodynamic parameters (all P<0.05). Conclusion:The right ACA and left PCA PSV are significantly associated with dementia in rural middle-aged and elderly people, and are expected to serve as non-invasive early screening biomarkers for cognitive impairment in rural population.

Objective To explore the effect of humanized nursing intervention on anxiety,depression and prognosis of maintenance hemodialysis patients.Methods A total of 90 patients with maintenance hemodialysis who were treated in our hospital were randomly divided into two groups,with 45 cases in each group.Patients in the control group were given routine care,and the patients in the observation group were given humanized nursing intervention on the basis of the control group.The psychological status (SAS,SDS) and quality of life of the two groups were compared and analyzed.Results After nursing,the scores of SDS and SAS were decreased than nursing before,and observation group was better than the control group (P ＜ 0.05).The compliance rate of the observation group was significantly higher than that of the control group (97.8％ vs.80.0％,P ＜ 0.05).After nursing intervention,the energy,physical effect,affective effect,social function,emotional status and overall health score of the observation group were significantly higher than intervention before,and these scores were significantly higher than that of the control group,and the differences were statistically significant (P ＜ 0.05).Conclusion Humanized nursing intervention for the maintenance of hemodialysis patients can significantly alleviate anxiety,depression and other negative emotions,establish a harmonious and stable nurse-patient relationship,and improve the prognosis and quality of life.

Objective To explore the effect of humanized nursing intervention on anxiety,depression and prognosis of maintenance hemodialysis patients.Methods A total of 90 patients with maintenance hemodialysis who were treated in our hospital were randomly divided into two groups,with 45 cases in each group.Patients in the control group were given routine care,and the patients in the observation group were given humanized nursing intervention on the basis of the control group.The psychological status (SAS,SDS) and quality of life of the two groups were compared and analyzed.Results After nursing,the scores of SDS and SAS were decreased than nursing before,and observation group was better than the control group (P ＜ 0.05).The compliance rate of the observation group was significantly higher than that of the control group (97.8％ vs.80.0％,P ＜ 0.05).After nursing intervention,the energy,physical effect,affective effect,social function,emotional status and overall health score of the observation group were significantly higher than intervention before,and these scores were significantly higher than that of the control group,and the differences were statistically significant (P ＜ 0.05).Conclusion Humanized nursing intervention for the maintenance of hemodialysis patients can significantly alleviate anxiety,depression and other negative emotions,establish a harmonious and stable nurse-patient relationship,and improve the prognosis and quality of life.

OBJECTIVETo investigate spontaneous metastasis, micrometastasis and genetic stability in human breast carcinoma xenografts in nude mice.

METHODSIntact tissue from surgical specimens from breast carcinoma patients was xenografted into nude mice and transplanted from generation to generation. Cells from the xenografts were cultured in vitro and retransplanted into nude mice. Microsatellite DNA in the genome of human breast carcinomas, xenotransplanted tumors and metastases in nude mice were analyzed at three microsatellite loci.

RESULTSThe tumorigenicity of orthotopic xenotransplantation was 88.6% (31/35), with a metastatic rate of 41.9% (13/31). Cells from xenotransplants were successfully cultured in vitro. The taking rate of retransplantation into nude mice and the spontaneous lung metastasis rate were both 100% (10/10). Microsatellite DNA sequences in the genome of xenotransplanted tumors and metastases in nude mice were identical with that of the original human breast carcinoma at three microsatellite loci.

CONCLUSIONSTumorigenicity and metastatic potential can be improved in human breast carcinoma xenografts using intact fresh tumor tissue and orthotopic grafts. Xenotransplanted tumors and tumors after serial passage maintained the genetic stability. The detection of microsatellite DNA may identify micrometastases in a nude mouse model.

Aneuploidy ; Animals ; Breast Neoplasms ; genetics ; pathology ; Cell Division ; Female ; Humans ; Mammary Neoplasms, Experimental ; genetics ; pathology ; Mice ; Mice, Nude ; Microsatellite Repeats ; Neoplasm Metastasis ; Neoplasm Transplantation ; Time Factors ; Transplantation, Heterologous ; Tumor Cells, Cultured

During 2 years,a 6-year-old girl was hospitalized for 2 times with recurrent onset of episodes of vomiting,weakness and fever after eating dessert at the Department of Neurology & Endocrine Pediatrics,the Affiliated Hospital of Qingdao University.The arterial blood gas analysis revealed severe hypoglycemia,lacticacidemia and metabolic acidosis,the urine ketone body was positive.After intravenous infusion of glucose,bicarbonate and antibiotics,there was a dramatic clinical improvement in a short time.Physical examination showed tachypnea and mild hepatomegaly,and she had normal physical and mental development.The laboratory findings revealed transient hyperuricacidemia.Urine organic acids analysis repeatedly showed an elevation of lactic acid,ketone and glycerol.Glyceroluria was a very distinctive trait.The literatures in PubMed was searched with glyceroluria as keyword.Three related diseases were identified:FBPase deficiency,glycerol kinase (GK) deficiency and complex GK deficiency.Further reading of related literatures to understand the characteristics of diseases and laboratory tests,the clinical diagnosis of GK deficiency and complex GK deficiency was excluded.The mutation analysis of FBPase gene (FBP1) was performed by Sanger sequencing and a novel compound heterozygous mutations of c.355G ＞A and c.960delG was discovered.Full analysis of disease-related traits and targeted gene testing is one of the effective methods for accurate diagnosis and treatment of inherited metabolic disorders.