Pasteurella multocida(Pm)type A is an important pathogen responsible for respiratory diseases,such as bovine pneumonia,which seriously restricts the development of cattle industry in China.Currently,the prevention of Pm infection-related diseases primarily relies on vaccination in production.However,the diverse Pm serotypes result in inadequate cross-immunological protection from vaccines.Therefore,it is of great significance to develop vaccines with cross-protection for the prevention and control of Pm infectious diseases.The previous studies conducted by our team have demonstrated that PmCQ2△cra exhibits a strong immune protective effect against Pm type A(PmA),Pm type B(PmB),and Pm type F(PmF).Transcriptomic sequencing results suggest that the cross-immunoprotective effect of PmCQ2△cra may be attributed to high expression levels of bacterial surface protective antigens.Consequently,four putative immune protective antigens,namely PmCQ2-5175,PmCQ2-6290,PmCQ2-0275 and PmCQ2-2640,were screened through bioin-formatics analysis in this study.Subunit vaccines formulated with these potential antigenic proteins exhibited protective efficacy of 62.5％,25％,12.5％and 10％against PmA-infected mice,respectively.Importantly,PmCQ2-5175,one of the most protective single-component antigen vac-cines,demonstrating a 75％cross-protection against PmB infection in mice.Furthermore,the pro-tective efficacy of the PmCQ2-5175 protein screened in this study was superior to that of the previ-ously reported Pm antigen protein plpE.Moreover,the fusion expression protein PmCQ2-5175-PLPE exhibited better protective effects against PmA compared to a single protein.The findings of this study will establish a theoretical foundation for the advancement of Pm subunit vaccines with broad-spectrum immune protection.

Pasteurella multocida(Pm)type A is an important pathogen responsible for respiratory diseases,such as bovine pneumonia,which seriously restricts the development of cattle industry in China.Currently,the prevention of Pm infection-related diseases primarily relies on vaccination in production.However,the diverse Pm serotypes result in inadequate cross-immunological protection from vaccines.Therefore,it is of great significance to develop vaccines with cross-protection for the prevention and control of Pm infectious diseases.The previous studies conducted by our team have demonstrated that PmCQ2△cra exhibits a strong immune protective effect against Pm type A(PmA),Pm type B(PmB),and Pm type F(PmF).Transcriptomic sequencing results suggest that the cross-immunoprotective effect of PmCQ2△cra may be attributed to high expression levels of bacterial surface protective antigens.Consequently,four putative immune protective antigens,namely PmCQ2-5175,PmCQ2-6290,PmCQ2-0275 and PmCQ2-2640,were screened through bioin-formatics analysis in this study.Subunit vaccines formulated with these potential antigenic proteins exhibited protective efficacy of 62.5％,25％,12.5％and 10％against PmA-infected mice,respectively.Importantly,PmCQ2-5175,one of the most protective single-component antigen vac-cines,demonstrating a 75％cross-protection against PmB infection in mice.Furthermore,the pro-tective efficacy of the PmCQ2-5175 protein screened in this study was superior to that of the previ-ously reported Pm antigen protein plpE.Moreover,the fusion expression protein PmCQ2-5175-PLPE exhibited better protective effects against PmA compared to a single protein.The findings of this study will establish a theoretical foundation for the advancement of Pm subunit vaccines with broad-spectrum immune protection.

Objective:To retrospectively analyze the clinical characteristics of 193 Chinese patients with McCune-Albright syndrome (MAS).Methods:By using keywords " McCune-Albright syndrome", " Albright syndrome", or " fibrous dysplasia " as the search terms, 193 cases of MAS reported in China from January 1990 to November 2022 from the Wanfang data, CNKI, VIP, PubMed, and Embase databases were obtained, and their clinical data was retrospectively analyzed. Intergroup comparisons were carried out by using t test, Mann-Whitney U test, and χ2 test. Results:The 193 MAS patients had included 42 males and 151 females, with the median first-visit age of females being younger than males. The typical triad group had accounted for 46.1% of patients, and the middle first-visit and diagnosis age was younger than the atypical group. The primary reason for first-visit in males of MAS was fibrous dysplasia (FD), whilst that in females of MAS was peripheral precocious puberty (PPP). FD has occurred in 84.5% of the patients, with an average age of onset age being 6.1 years old, and 90% was ≤ 16 years of age. Endocrine hyperfunction was found in 79.3% of the patients, with a higher proportion in females compared with males ( P<0.05). Pituitary involvement was seen in 21.8% of the patients, and the incidence of craniofacial FD and cranial nerve compression was significantly higher in those with elevated growth hormone (GH) than without ( P<0.05). Café-au-Lait Spots were noted in 86.5% of the patients, and 28.3% (28/99) had located on the different side of FD. Conclusion:Most MAS patients had atypical manifestations and multi-systemic involvement. It is more common and occurs earlier in females. The most common reasons for initial diagnosis in male and female patients were FD and PPP, respectively. Patients with elevated GH should be examined for cranial nerve compression.

OBJECTIVE: To assess the efficacy and safety of aromatase inhibitors (AIs) combined growth hormone in treatment of adolescent boys with short stature. METHODS: One hundred and fifty-one short stature pubertal boys with age of 10-14 years and bone age of 13-15 years, who were admitted to the Department of Pediatrics, the First Affiliated Hospital, Zhejiang University School of Medicine, were included in this trial. According to their own or parents' intention, the children were divided into recombinant human growth hormone (rhGH)+AI group ( =108) and rhGH group ( =43). All children were injected subcutaneously with rhGH 0.15-0.2 IU·kg ·d , and those in rhGH+AI group were additionally given 2.5 mg/d letrozole or 1 mg/d anastrozole, orally for 12 months or longer. The children were followed-up every 3 months. During the follow-up visit, the predicted adult height (PAH), sex hormone level, glucose and lipid metabolism, and other indicators were measured, and adverse reactions were monitored. RESULTS: After intervention, there were significant differences in ΔBA(bone age)/ΔCA(chronological age), ΔHtSDS (height standard deviation score based on bone age)and ΔPAH between rhGH+AI group and the rhGH group( < 0.05 or < 0.01). During follow-up, 63.9%of the children in the rhGH+AI group had elevated uric acid and 51.9%had decreased high-density lipoprotein (HDL); 25.9%showed severe acne, excitement, hyperactivity and irritability, 11.1%had knee pain; 4.6%had fracture; 2.8%had mild renal dysfunction; 1.9%had inactivity, drowsiness, memory loss and performance decline; 1.9%showed mild abnormal liver function; 0.9%showed impaired fasting glucose; 0.9%showed granulocytopenia. In the rhGH group, 11.6%of the children presented with knee pain and 2.3%with impaired fasting glucose. CONCLUSIONS AI combined with rhGH can delay the growth of BA and effectively improve the PAH of adolescent boys with larger bone age. However, the occurrence of adverse reactions of AI should be closely monitored during treatment.
Adolescent ; Aromatase Inhibitors ; therapeutic use ; Body Height ; Child ; Growth Disorders ; Human Growth Hormone ; Humans ; Male ; Recombinant Proteins

Objective To explore the gene mutation characteristics and detailed clinical presentations of hyperglycemia caused by GCK mutations in 10 patients.Methods The clinical and follow-up data of 10 patients with hyperglycemia caused by mutation of GCK gene were reviewed.The patients were ascertained between January 1,2014 and August 31,2018 at the Department of Pediatrics,the First Affiliated Hospital of Zhejiang University and Ningbo Women & Children's Hospital.Clinical data were collected,including age,gender,main complaint,family history,fasting blood glucose,fasting blood insulin,2-hour blood glucose,2-hour blood insulin after oral glucose tolerance test,glycosylated hemoglobin,anti-glutamic acid decarboxylase antibody and body mass index.Mutations of GCK gene were detected by Sanger sequencing or high-throughput sequencing of diabetes-related genes in the patients and their family members.Results There were ten patients,8 of them were male,2 were female.The ages at diagnosis varied between 4.7 to 12.3 years.The patients usually did not have obvious clinical symptoms of diabetes mellitus.Most of them were unexpectedly found to have hyperglycemia and with impaired glucose metabolism in three consecutive generations.The fasting blood glucose of patients was 6.8-7.7 mmol/L,2-hour postprandial blood glucose was 7.8-11.6 mmol/L.Fasting blood insulin was 0.5-8.5 mU/L,glucose tolerance test results showed that 2 h postprondial blood insulin was 1.3-55.4 mU/L.The level of glycosylated hemoglobin was 6.1％-6.8％.Anti-glutamic acid decarboxylase antibody was negative in all patients.The GCK mutations identified in patients and one of their parents were located at exon5 (4 cases),exon9 (2 cases),exon2 (1 case),exon4 (1 case),exon6 (1 case) and exon7 (1 case).Conclusions Most of the hyperglycemia patients caused by GCK mutations did not have typical clinical symptoms of diabetes.The fasting blood glucose was slightly elevated.Abnormal glucose tolerance test results were found in all 10 patients.Three consecutive generations of family had impaired glucose metabolism.GCK mutations located at exon 5 were common in 10 cases.There was no correlation between type of mutations and plasma glucose levels in domestic and international researches.When fasting glucose was found abnormal in clinic,a complete family history should be taken and the GCK gene should be sequenced to confirm the diagnosis in time.

Objective To study the AAAS gene mutations in a child with autosomal recessive Allgrove syndrome. Methods Clinical data were collected and blood samples were obtained from the proband of Allgrove syndrome and her parents. Genomic DNA was extracted and sequenced by PCR amplification. Subclone sequencing was performed to validate the gene mutations. The disease-causing potentials of mutation genes were evaluated by the Mutation Taster, and the target protein tertiary structure was modelled by the Swiss Model. Results A new heterozygous insertion mutation(c. 1347_1348insG) of exon 15 in the proband was identified and firstly reported. Other two reported mutations were detected, which were the heterozygous mutation c. 688C>T in the patient and her mother, and the homozygous mutation c. 855C>T in the proband and her parents. In addition, it was confirmed that the novel compound heterozygous mutations(c. 688C>T, c. 1347_1348insG) in the AAAS gene of the proband were pathogenic mutation locus. Conclusion The heterozygous mutation(c. 1347_1348insG) of AAAS gene was firstly reported. In case of the patients being clinically misdiagnosed, related-gene detection should be performed for the patients who were diagnosed with primary adrenal insufficiency during the period of infants and young childhood.

OBJECTIVETo report on two cases affected with immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX).

METHODSTwo unrelated Chinese infants affected with IPEX were investigated. Case 1 was a 4-month-old boy with neonatal diabetes and severe enteropathy. Case 2 was a 6-day newborn boy with neonatal diabetes and ketoacidosis. DNA samples of the two infants and their parents were sequenced for FOXP3 gene mutations. Suspected mutations were verified among 100 unrelated healthy controls. The function of mutations was predicted with bioinformatics software.

RESULTSBoth infants had onset of the disease during neonatal period, and manifested insulin-dependent diabetes mellitus, persistent diarrhea, eczema and malnutrition. In case 1, a novel splice site mutation was identified in intron 9 (c.967+3A>T) of the FOXP3 gene, for which his mother was a carrier. For case 2, a missense mutation (c.1150G>A) was detected in exon 11 of the FOXP3 gene, for which his mother was also a carrier. The IVS9 c.967+3A mutation was not detected among the 100 healthy controls. As predicted with Human Splicing Finder software, the c.967+3A>T mutation may influence the splicing of mRNA and affect the function of protein.

CONCLUSIONBoth cases had typical clinical manifestation of the IPEX syndrome, among whom a novel splice site mutation (IVS9 c.967+3A>T) and a missense mutation (c.1150G>A) of the FOXP3 gene were identified. The clinical manifestation of the IPEX syndrome may be variable and the mortality is high. FOXP3 gene sequencing is recommended when insulin-dependent diabetes mellitus is diagnosed during the neonatal period.

A 12-year-old girl presented with a history of cervical mass, and one week of throat discomfort and dyspnea. Five years ago, the patient was diagnosed as Hashimoto's thyroiditis and hyperthyroidism; she received antithyroid drug treatment, but the result was not satisfactory. B-ultrasonic showed that the size of thyroid gland was 8.1 cm×3.2 cm in the left and 8.2 cm×4.8 cm in the right. After iodine 131 combined with radiofrequency ablation (RFA) treatment, throat discomfort and recumbent breathing difficulties disappeared, and B-ultrasonic showed that the size of thyroid reduced to 2.3 cm×1.7 cm (left) and 2.8 cm×2.0 cm (right). No recurrence was observed during the two and a half years of follow-up.
Ablation Techniques ; methods ; Child ; Dyspnea ; etiology ; therapy ; Female ; Goiter ; complications ; diagnostic imaging ; pathology ; therapy ; Hashimoto Disease ; therapy ; Humans ; Hyperthyroidism ; therapy ; Iodine Radioisotopes ; therapeutic use ; Radio Waves ; therapeutic use ; Ultrasonography

Objective To investigate association of the paired box 4 (PA X 4) gene rs3824004 (574C>A; R192S) and rs2233580 (575G>A; R192H) polymorphism with obesity and metabolic markers in children and adolescents. Methods A total of 103 obese children were randomly selected, and an average age was (10.82±2.57) years, and body mass index (BMI) was (26.82±4.57) kg/m2. At the same period, 100 normal weight children were selected as the control group, and an average age of (10.60±2.84) years, and BMI was (16.79±2.13) kg/m2. The blood pressure, physical measurements, and blood metabolic parameters were measured and compared. The oral glucose tolerance test (OGTT) and insulin release test were performed in the obesity group. The homeostasis model insulin resistance index (HOMA-IR) and the overall insulin sensitivity index (WBISI) were calculated. PA X 4 rs3824004 and rs2233580 polymorphism were detected by PCR.The differences of allele frequency and genotype frequency of polymorphic loci were analyzed, and the correlation between different genotypes and metabolic indexes was analyzed. Results The height, weight, BMI, systolic blood pressure, diastolic blood pressure, waist circumference, hip circumference, waist to height ratio (WHtR), fasting blood glucose (FPG), total cholesterol (TC), low density lipoprotein (LDL), triacylglycerol (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the obesity group were significantly higher than those in the control group, and the high density lipoprotein (HDL) was significantly lower than that in the control group (all P<0.05). The frequency of gene distribution was in accordance with the Hard-Weinberg balance. The frequencies of A allele of rs3824004 in obesity and control groups were 4.9% and 5.0%, respectively, and the frequencies of CA genotype were 9.7% and 10.0%, respectively, and there was no significant difference between two groups (P>0.05). The frequency of GA allele of rs2233580 in obesity group was 25.2%, which was significantly higher than that in control group (P<0.05). The BMI and waist in rs2233580 GA genotype were significantly higher than those in GG genotype (all P <0.05). However,logistic regression analysis showed that there was no correlation between PA X 4 rs2233580 genotype and metabolic markers (all P>0.05).There were no significantly differences in HOMA-IR and WBISI among different genotypes of PA X 4 rs2233580 in obesity group(all P>0.05).Conclusions PA X 4 rs2233580 affects children's BMI and waist circumference and may be involved in the development of childhood obesity, but it is not an independent risk factor for obesity in children and adolescents.

Objective To investigate the clinical characteristics of twins with thyroxine binding globulin (TBG) deficiency and to find SERPINA7 gene mutations.Method Data(2015) related to clinical characteristics,serum biochemistry,gene mutations and pedigree of two children with TBG deficiency were collected in the First Affiliated Hospital of College of Medicine,Zhejiang University.The related literature was searched form China National Knowledge Infrastructure,Wanfang Data Knowledge Service Platform,National Center for Biotechnology Information and PubMed (up to December 2015) by using search terms "Thyroxine binding globulin deficiency,gene,mutation".Result Both patients were diagnosed as central hypothyroidism at the beginning and treated with L-thyroxine.Both of the identical twins of the triplet were observed for mutation in exon3,c.631 G ＞ A(p.A211T),a new mutation had not been reported,but their parents and another non-identical triplet brother were normal.Literature review showed that 23 foreign cases with SERPINA7 gene mutation had been reported,however,no Chinese with SERPINA7 gene mutation had been reported.Among reported cases it was shown that SERPINA7 gene mutations located in exon,intron,promoter and enhancer.Up to now,49 variants had been identified,41 of them located in the mutated genes.Including these two cases,patients with thyroxine binding globulin deficiency were characterized by reduced serum TH levels,but normal free TH and TSH and absence of clinical manifestations.Conclusion The new mutation of SERPINA7 gene c.631G ＞ A (p.A211T)is not transmitted via the known X chromosome linked heredity,and as the cases were test tube triplet infants,it is a de novo mutation.The serum thyroid function tests of TBG deficiency showed decreased TT4,TT3 and normal TSH and TBG deficiency is often misdiagnosed as central hypothyroidism.