Objective To investigate the levels of serum neuregulin-4(NRG4)and serum fatty acid binding protein 4(FABP4)in patients with diabetes retinopathy(DR)and their diagnostic value.Methods From A-pril 2020 to April 2023,a total of 84 DR patients admitted to Endocrine and Metabolic Disease Hospital Affili-ated to Shandong First Medical University were regarded as the observation group.Based on the type of retinal lesions,they were grouped into proliferative lesion group(35 cases)and non proliferative lesion group(49 ca-ses).In addition,84 patients with simple diabetes in the same period were regarded as the NDR group,and 84 healthy people who underwent physical examination in the same period were regarded as the control group.Enzyme linked immunosorbent assay was applied to detect serum levels of NRG4 and FABP4.Multivariate Logistic regression was applied to analyze the influencing factors of DR occurrence.Receiver operating charac-teristic(ROC)curve was applied to analyze the diagnostic value of serum NRG4 and FABP4 levels for DR.Results The levels of C-reactive protein(CRP),fasting plasma glucose(FPG),fasting insulin(FINS)and FABP4 in the observation group were higher than those in the NDR group and the control group(P<0.05),and the serum NRG4 level was lower than those in the NDR group and the control group(P<0.05).The ser-um NRG4 level in the proliferative lesion group was lower than that in the non proliferative lesion group(P<0.05),and the FABP4 level was higher than that in the non proliferative lesion group(P<0.05).NRG4 was a protective factor for the occurrence of DR(P<0.05),while FABP4 was a risk factor for the occurrence of DR(P<0.05).The area under the curve(AUC)of serum NRG4 and FABP4 for diagnosing DR was 0.850 and 0.866,and the AUC of combined diagnosis was 0.930.The efficacy of the combination of the two was su-perior to that of the individual diagnosis of serum NRG4 and FABP4(Zcombination-NRG4=3.467,Zcombination-FABP4=2.987,P=0.001,0.002).Conclusion The serum FABP4 level in DR patients is greatly increased,while the NRG4 level is greatly decreased.The combination of the two has high diagnostic value for DR.

Objective To compare the hematological parameters,biochemical profiles,histopathological characteristics,and cytokines associated with lipid metabolism in 8-weeks-old specific pathogen-free(SPF)grade and germ-free(GF)golden hamsters.Methods Twenty 8-weeks-old SPF grade and GF golden hamsters,with equal numbers of males and females,were utilized in this study.Serum cytokines associated with routine blood parameters,blood biochemistry,and lipid metabolism were quantified using automated hematology and biochemical analyzers,and enzyme-linked immunosorbent assay kits.The cecum,small intestine,lung,spleen,forestomach,and glandular stomach were examined by histopathology.Results We compared the result between SPF grade and GF golden hamsters at 8 weeks of age.Regarding hematological parameters,females showed significant differences(P＜0.05)in white blood cell count,hemoglobin concentration,mean corpuscular hemoglobin(MCH),and mean platelet volume between the two groups,and highly significant differences(P＜0.001)in MCH concentration(MCHC),platelet distribution width(PDW),and lymphocyte count(LYM).Males showed significant differences(P＜0.05)in mean corpuscular volume,MCH,LYM,neutrophil count,basophil count,and basophil percentage,and highly significant differences(P＜0.001)in MCHC,PDW,lymphocyte percentage,and neutrophil percentage.In terms of biochemical parameters,females showed significant differences(P＜0.05)in low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol,fasting insulin(FINS),and glycosylated serum protein(GSP),and highly significant differences(P＜0.001)in triglycerides(TG).Males showed significant differences(P＜0.05)in GSP levels and highly significant differences(P＜0.001)in total cholesterol,TG,LDL-C,FINS,and C-peptide levels.For serum cytokines related to lipid metabolism,females showed significant differences(P＜0.05)in interleukin(IL)-10,adiponectin(ADP),and IL-6 levels,and highly significant differences(P＜0.001)in high-sensitivity C-reactive protein(hs-CRP).Males showed significant differences(P＜0.05)in hs-CRP levels and highly significant differences(P＜0.001)in ADP and tumor necrosis factor-alpha levels.Hematoxylin/eosin staining showed that the cecal muscle layer was thinner and the number of crypts attached to the mucous membrane was reduced in GF compared with SPF grade golden hamsters.In addition,the ileocele was enlarged and the number of goblet cells was increased,the alveolar septum was widened,immune cells in the white pulp of the spleen were increased,and the blood content in the blood sinuses was increased.There was also thinning of the anterior gastric mucosa and the basophilic strength of the glandular gastric tube gland was weakened.Conclusions This study established the differences in routine blood parameters,blood biochemistry indicators,histopathological features,and cytokines associated with lipid metabolism between 8-weeks-old SPF grade and GF golden hamsters,to establish preliminary reference ranges.

Objective To compare the hematological parameters,biochemical profiles,histopathological characteristics,and cytokines associated with lipid metabolism in 8-weeks-old specific pathogen-free(SPF)grade and germ-free(GF)golden hamsters.Methods Twenty 8-weeks-old SPF grade and GF golden hamsters,with equal numbers of males and females,were utilized in this study.Serum cytokines associated with routine blood parameters,blood biochemistry,and lipid metabolism were quantified using automated hematology and biochemical analyzers,and enzyme-linked immunosorbent assay kits.The cecum,small intestine,lung,spleen,forestomach,and glandular stomach were examined by histopathology.Results We compared the result between SPF grade and GF golden hamsters at 8 weeks of age.Regarding hematological parameters,females showed significant differences(P＜0.05)in white blood cell count,hemoglobin concentration,mean corpuscular hemoglobin(MCH),and mean platelet volume between the two groups,and highly significant differences(P＜0.001)in MCH concentration(MCHC),platelet distribution width(PDW),and lymphocyte count(LYM).Males showed significant differences(P＜0.05)in mean corpuscular volume,MCH,LYM,neutrophil count,basophil count,and basophil percentage,and highly significant differences(P＜0.001)in MCHC,PDW,lymphocyte percentage,and neutrophil percentage.In terms of biochemical parameters,females showed significant differences(P＜0.05)in low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol,fasting insulin(FINS),and glycosylated serum protein(GSP),and highly significant differences(P＜0.001)in triglycerides(TG).Males showed significant differences(P＜0.05)in GSP levels and highly significant differences(P＜0.001)in total cholesterol,TG,LDL-C,FINS,and C-peptide levels.For serum cytokines related to lipid metabolism,females showed significant differences(P＜0.05)in interleukin(IL)-10,adiponectin(ADP),and IL-6 levels,and highly significant differences(P＜0.001)in high-sensitivity C-reactive protein(hs-CRP).Males showed significant differences(P＜0.05)in hs-CRP levels and highly significant differences(P＜0.001)in ADP and tumor necrosis factor-alpha levels.Hematoxylin/eosin staining showed that the cecal muscle layer was thinner and the number of crypts attached to the mucous membrane was reduced in GF compared with SPF grade golden hamsters.In addition,the ileocele was enlarged and the number of goblet cells was increased,the alveolar septum was widened,immune cells in the white pulp of the spleen were increased,and the blood content in the blood sinuses was increased.There was also thinning of the anterior gastric mucosa and the basophilic strength of the glandular gastric tube gland was weakened.Conclusions This study established the differences in routine blood parameters,blood biochemistry indicators,histopathological features,and cytokines associated with lipid metabolism between 8-weeks-old SPF grade and GF golden hamsters,to establish preliminary reference ranges.

Objective A sterile golden hamster model was established by cesarean section purification.Methods SPF-grade donor female golden hamsters were selected,and males and females were mated 1∶1 and separated after mating.The cage time of the surrogate mothers was 1 week earlier than that of the donor mothers.Parturient golden hamsters underwent hysterectomy on a sterilized workbench,and the uteruses were transferred into isolation kits and stripped.To obtain sterile milk for milk replacement,sterile ICR mice and sterile SD rats were used.After successful separation,the hamsters were transferred to isolation kits to prepare for feeding.The sterility status of the feeding isolation kits was tested monthly.Results Three caesarean sections were performed,but the first and second lactations failed.The third milk replacement was successful,and 18 young hamsters were obtained with survival rates of 88%and 66%after weaning.All hamsters were quality tested by GB/T 14926.41-2001.Conclusions Using a cesarean section purification technique and sterile ICR mice and SD rats for microbial-free milk replacement,a sterile golden hamster model was obtained.

Objective:To analyze the factors affecting delayed chemotherapy in children with Burkitt lymphoma (BL) and their influence on prognosis.Methods:Retrospective cohort study. Clinical data of 591 children aged ≤18 years with BL from May 2017 to December 2022 in China Net Childhood Lymphoma (CNCL) was collected. The patients were treated according to the protocol CNCL-BL-2017. According to the clinical characteristics, therapeutic regimen was divided into group A, group B and group C .Based on whether the total chemotherapy time was delayed, patients were divided into two groups: the delayed chemotherapy group and the non-delayed chemotherapy group. Based on the total delayed time of chemotherapy, patients in group C were divided into non-delayed chemotherapy group, 1-7 days delayed group and more than 7 days delayed group. Relationships between delayed chemotherapy and gender, age, tumor lysis syndrome before chemotherapy, bone marrow involvement, disease group (B/C group), serum lactate dehydrogenase (LDH) > 4 times than normal, grade Ⅲ-Ⅳ myelosuppression after chemotherapy, minimal residual disease in the interim assessment, and severe infection (including severe pneumonia, sepsis, meningitis, chickenpox, etc.) were analyzed. Logistic analysis was used to identify the relevant factors. Kaplan-Meier method was used to analyze the patients' survival information. Log-Rank was used for comparison between groups.Results:Among 591 patients, 504 were males and 87 were females, the follow-up time was 34.8 (18.6,50.1) months. The 3-year overall survival (OS) rate was (92.5±1.1)%,and the 3-year event-free survival (EFS) rate was (90.5±1.2)%. Seventy-three (12.4%) patients were in delayed chemotherapy group and 518 (87.6%) patients were in non-delayed chemotherapy group. The reasons for chemotherapy delay included 72 cases (98.6%) of severe infection, 65 cases (89.0%) of bone marrow suppression, 35 cases (47.9%) of organ dysfunction, 22 cases (30.1%) of tumor lysis syndrome,etc. There were 7 cases of chemotherapy delay in group B, which were seen in COPADM (vincristine+cyclophosphamide+prednisone+daunorubicin+methotrexate+intrathecal injection,4 cases) and CYM (methotrexate+cytarabine+intrathecal injection,3 cases) stages. There were 66 cases of chemotherapy delay in group C, which were common in COPADM (28 cases) and CYVE 1 (low dose cytarabine+high dose cytarabine+etoposide+methotrexate, 12 cases) stages. Multinomial Logistic regression analysis showed that the age over 10 years old ( OR=0.54,95% CI 0.30-0.93), tumor lysis syndrome before chemotherapy ( OR=0.48,95% CI 0.27-0.84) and grade Ⅲ-Ⅳ myelosuppression after chemotherapy ( OR=0.55,95% CI 0.33-0.91)were independent risk factors for chemotherapy delay.The 3-year OS rate and the 3-year EFS rate of children with Burkitt lymphoma in the delayed chemotherapy group were lower than those in the non-delayed chemotherapy group ((79.4±4.9)% vs. (94.2±1.1)%, (80.2±4.8)% vs. (92.0±1.2)%,both P<0.05). The 3-year OS rate of the group C with chemotherapy delay >7 days (42 cases) was lower than that of the group with chemotherapy delay of 1-7 days (22 cases) and the non-delay group (399 cases) ((76.7±6.9)% vs. (81.8±8.2)% vs. (92.7±1.3)%, P=0.002).The 3-year OS rate of the chemotherapy delay group (9 cases) in the COP (vincristine+cyclophosphamide+prednisone) phase was lower than that of the non-chemotherapy delay group (454 cases) ((66.7±15.7)% vs. (91.3±1.4)%, P=0.005). Similarly, the 3-year OS rate of the chemotherapy delay group (11 cases) in the COPADM1 phase was lower than that of the non-chemotherapy delay group (452 cases) ((63.6±14.5)% vs. (91.5±1.3)%, P=0.001). Conclusions:The delayed chemotherapy was related to the age over 10 years old, tumor lysis syndrome before chemotherapy and grade Ⅲ-Ⅳ myelosuppression after chemotherapy in pediatric BL. There is a significant relationship between delayed chemotherapy and prognosis of BL in children.

Fetal STR typing of aborted tissue has long been a major problem in forensic DNA.Especially for the first trimester abortion tissue,it is difficult to isolate the embryonic components by histomorphological means,resulting in the inability to accurately obtain the STR typing of the fetus.The mixed STR typing results of mother and fetus can provide a key basis for the identification of suspects in cases of rape-induced pregnancy.In this study,next generation sequencing was used to successfully detect mixed STR typing of mother and suspected fetus or single STR typing of suspected fetus in 4 rape-induced early pregnancy abortion tissues.Combined with Y-STR and flank sequence information,it provides a more comprehensive and reliable genetic basis for the identification of suspects.

Objective:To explore the clinical and genetic features of two Chinese pedigrees affected with Autosomal dominant intellectual developmental disorder 49 (MRD49).Methods:Two MRD49 pedigrees which were admitted to the Children′s Hospital Affiliated to Shandong University respectively on January 28, 2021 and November 10, 2022 were selected as the study subjects. Clinical data of the two pedigrees were collected and analyzed. Genomic DNA was extracted from peripheral blood samples of the probands and their family members. The probands were subjected to mutational analysis by high-throughput sequencing. Candidate variants were validated using real-time fluorescence quantitative PCR (q-PCR) or Sanger sequencing and bioinformatic analysis. This study was approved by the Medical Ethics Committee of the Children′s Hospital Affiliated to Shandong University (No. SDFE-IRB/T-2022002).Results:Proband 1 had presented with language delay, motor retardation and intellectual disability, and his maternal grandmother, mother, aunt and cousin all had various degrees of intellectual disability. Sequencing results showed that proband 1 had deletion of exons 3 ~ 7 of the TRIP12 gene. q-PCR verification showed that his mother, aunt, maternal grandmother and cousin had all harbored the same deletion. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP1). Proband 2, who had mainly presented with language delay, motor retardation and intellectual disability, and was found to harbor a heterozygous c.3010C>T (p.Arg1004*) variant of the TRIP12 gene, which was verified to be de novo in origin. Based on the guidelines from the ACMG, the variant was classified as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:This study had diagnosed two MRD49 families through high-throughput sequencing. Above findings have enriched the phenotypic and mutational spectrum of MRD49 in China, which has also facilitated genetic counseling for the two pedigrees.

Objective To evaluate the effects of fecal status and transplantation method on the intestinal flora structure after fecal microbiota transplantation in germ-free mice.Methods Thirty-six C57BL/6 mice were divided randomly into three groups:fresh fecal gavage transplantation(group A),frozen fecal gavage transplantation(group B),and frozen fecal rectal transplantation groups(group C).Feces were collected at 2,4 and 6 weeks after transplantation.All mice were sacrificed at 6 weeks to obtain the contents of the small and large intestines.The structure and function of the gut microbiota and dynamic trends in microbial changes were analyzed by 16 S rRNA sequencing.Results The diversity of the small intestine microbes in both the group A and group C were similar to those in the group B,according to α-diversity analysis(P＞0.05),but the diversity of large intestine microbes was significantly increased(P＜0.001).According toβ-diversity analysis,small intestine samples from the group A and group B clustered in the same area,indicating that the microbial community compositions were similar(P＞0.05),but samples from the large intestine were distributed in different areas,showing significant differences(P＜0.001).Small and large intestine samples from the group B and group C were distributed in different areas,with significant differences(P＜0.001).Linear discriminant analysis effect size showed that Bacteroidota were relatively dominant in the group A,while Verrucomicrobiota and Proteobacteria were relatively dominant in the group B and Firmicutes were relatively dominant in the group C.Functional prediction using PICRUSt2 software showed that neither fecal status nor the method of transplantation affected the functions of the microbial community.Conclusions Both fresh fecal gavage and frozen fecal rectal transplantation can enhance the microbial diversity of the large intestine,compared with frozen fecal gavage transplantation.Fecal status does not affect the gut function and colonization trends of the microbiota,whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.

Objective To evaluate the effects of fecal status and transplantation method on the intestinal flora structure after fecal microbiota transplantation in germ-free mice.Methods Thirty-six C57BL/6 mice were divided randomly into three groups:fresh fecal gavage transplantation(group A),frozen fecal gavage transplantation(group B),and frozen fecal rectal transplantation groups(group C).Feces were collected at 2,4 and 6 weeks after transplantation.All mice were sacrificed at 6 weeks to obtain the contents of the small and large intestines.The structure and function of the gut microbiota and dynamic trends in microbial changes were analyzed by 16 S rRNA sequencing.Results The diversity of the small intestine microbes in both the group A and group C were similar to those in the group B,according to α-diversity analysis(P＞0.05),but the diversity of large intestine microbes was significantly increased(P＜0.001).According toβ-diversity analysis,small intestine samples from the group A and group B clustered in the same area,indicating that the microbial community compositions were similar(P＞0.05),but samples from the large intestine were distributed in different areas,showing significant differences(P＜0.001).Small and large intestine samples from the group B and group C were distributed in different areas,with significant differences(P＜0.001).Linear discriminant analysis effect size showed that Bacteroidota were relatively dominant in the group A,while Verrucomicrobiota and Proteobacteria were relatively dominant in the group B and Firmicutes were relatively dominant in the group C.Functional prediction using PICRUSt2 software showed that neither fecal status nor the method of transplantation affected the functions of the microbial community.Conclusions Both fresh fecal gavage and frozen fecal rectal transplantation can enhance the microbial diversity of the large intestine,compared with frozen fecal gavage transplantation.Fecal status does not affect the gut function and colonization trends of the microbiota,whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.

OBJECTIVE: To explore the genetic etiology of a child with Hypomagnesemia, epilepsy and mental retardation syndrome (HSMR). METHODS: A child who was admitted to the Children's Hospital of Shandong University on July 9, 2021 due to repeated convulsions for 2 months was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and his pedigree members were collected for the extraction of genomic DNA. Whole exome sequencing was carried out, and candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The child, a 1-year-and-7-month-old male, had presented with epilepsy and global developmental delay. Serological testing revealed that he has low serum magnesium. Genetic testing showed that the child has harbored a heterozygous c.1448delT (p.Val483GlyfsTer29) variant of the CNNM2 gene, which was de novo in origin. The variant has caused substitution of the Valine at position 483 by Glycine and formation of a termination codon after 29 amino acids at downstream. As predicted by Swiss-Model online software, the variant may alter the protein structure, resulting in a truncation. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1448delT (p.Val483GlyfsTer29) was predicted as a pathogenic variant (PVS1+PS2+PM2_Supporting+PP4). CONCLUSION The heterozygous c.1448delT variant of the CNNM2 gene probably underlay the HSMR in this child. Above finding has enriched the phenotype-genotype spectrum of the CNNM2 gene.
Humans ; Male ; Cation Transport Proteins ; Computational Biology ; Ethnicity ; Intellectual Disability/genetics* ; Magnesium ; Mutation ; Seizures/genetics* ; Infant