Objective:To analyze the clinical phenotype and genetic basis of a Chinese pedigree affected with Otopalatodigital syndrome type 1 (OPD1).Methods:A pedigree which was evaluated at the Department of Endocrinology, General Hospital of the Central Theater Command on December 3, 2020 was selected as the study subject. Clinical phenotype and genetic features of the proband were analyzed. Whole exome sequencing was employed to screen for genetic variants in the proband, and Sanger sequencing was used to verify the candidate variants in the proband′s mother, uncle, maternal aunt, and paternal aunt. Pathogenicity analysis was also conducted for the candidate variants.Results:The proband, a 16-year-old male, had shown distinctive facial features including mildly prominent eyebrows, down-slanting palpebral fissures, hypertelorism, and depressed nasal bridge. Additionally, he had clubbing of bilateral thumbs and big toes, and central type diabetes insipidus. Genetic sequencing revealed that he has harbored a heterozygous c. 586C>T (p.R196W) missense variant of the FLNA gene (NM_001110556.2), which was also carried by his mother and uncle. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as likely pathogenic (PM1+ PM2_Supporting+ PP2+ PP3+ PS4 Supporting). Conclusion:The heterozygous c. 586C>T (p.R196W) variant of the FLNA gene probably underlay the pathogenesis in this OPD1 family. The central type diabetes insipidus in the proband may represent a newly discovered phenotype of OPD1. Above finding has contributed crucial information for the comprehensive understanding of the clinical manifestations and pathogenic mechanisms of OPD1.

Objective To investigate the relationship between serum neutrophil extracellular trapps(NETs)levels and carotid intima-media thickness(CIMT)in patients with type 2 diabetes mellitus(T2DM).Methods A total of 112 newly diagnosed T2DM patients were selected from September 2022 to May 2023.Patients were divided into the T2DM group(T2DM,n=35),T2DM+the intima-media thickening group(CIMT,n=39)and T2DM+the atheromatous plaque group(PLA,n=38)according to the CIMT.And 50 cases of healthy people were selected as the normal control(NC)group in the same period.Results Compared with the NC group,the BMI,SBP,FPG,2 hPG,TC,and NETs increased in the T2DM group(P<0.05);The BMI,SBP,HbA1c,2 hPG,TG,CIMT,HOMA-IR,and NETs were higher in the CIMT group than in the T2DM group(P<0.05),while FPG,TC,and LDL-C were higher in the CIMT group than in the NC group(P<0.05).The SBP,HbA1c,2 hPG,LDL-C,CIMT,and NETs were higher in the PLA group than in the CIMT group(P<0.05),while BMI,FPG,TG,and HOMA-IR were higher in the PLA group than in the T2DM group(P<0.05).Spearman correlation analysis showed that CIMT in T2DM patients was positively correlated with NETs,smoking,BMI,SBP,FPG,2 hPG,HbA1c,LDL-C,TC,TG,UACR and HOMA-IR(P<0.05).Multiple linear regression analysis showed that NETs,HbA1c,LDL-C and TG were independent risk factors for CIMT in the T2DM patients.The analysis of the working characteristic curve of the subjects showed that the area under the curve of serum NETs for diagnosing T2DM combined with AS was 0.960,with sensitivity of 97.4%and specificity of 84.7%.Conclusion Serum NETs are associated with atherosclerosis in T2DM patients.

Objective:To investigate the effects of myeloid-derived growth factor(MYDGF) on inflammatory response and osteoclast differentiation of RAW264.7 cells.Methods:The RAW264.7 osteoclast precursor cells were cultured and treated with different concentrations of recombinant MYDGF protein(rMYDGF), and their cell viability was assessed using the MTT assay. RAW264.7 cells were induced with lipopolysaccharide(LPS) to induce inflammation, and the expression of inflammatory mediators and cell polarization were observed after intervention with rMYDGF. The RAW264.7 cells were induced for osteoclast differentiation using receptor activator of nuclear factor-κB ligand(RANKL), and rMYDGF was added for intervention. Osteoclast differentiation was evaluated by tartrate-resistant acid phosphatase(TRAP) staining. The osteoclast resorption pits and the number of actin rings(F-actin rings) were observed under a microscope. Reverse transcription PCR was performed to detect the expression of activated T cell nuclear factor 1(Nfatc1), cathepsin K(CTSK), and c-Fos genes during osteoclast differentiation. The protein phosphorylation levels of nuclear factor-κB(NF-κB) signaling pathway proteins were detected using Western blotting.Results:MTT assay showed that rMYDGF did not significantly inhibit the viability of RAW264.7 cell when the concentration was lower than 100 ng/mL. Moreover, rMYDGF inhibited the expression levels of inflammatory factors and M1 cell polarization after LPS stimulation. Compared with the control group, the number and area of TRAP positive cells, the number and area of bone resorption pit were decreased in rMYDGF intervention group respectively, as well as the area of the F-actin ring was reduced and its shape was incomplete after rMYDGF intervention. Furthermore, rMYDGF reduced the expression levels of osteoclast-specific marker genes and inhibited the phosphorylation of NF-κB signaling pathway protein IκBα during osteoclast differentiation.Conclusion:MYDGF inhibits the inflammatory response and osteoclast differentiation of RAW264.7 cells.

Objective:To investigate the effect of alogliptin on bone loss in ovariectomized(OVX)mice.Methods:For animal experiments, thirty 8-week-old C57BL/6J female mice were divided into Sham group, OVX group, and OVX+ alogliptin group. OVX+ alogliptin group were administered with alogliptin in a dosage of 20 mg·kg -1·d -1 by gavage, Sham and OVX groups with equivalent saline. After 12 weeks intervention, serum bone anabolism indicators were detected, and Micro CT and HE staining were used to observe and analyze the bone trabecular structure of femur and tibia in mice. For in vitro experiments, bone marrow mesenchymal stem cells(BMSCs)were incubated with 100 μmol/L alogliptin for osteoblast differentiation. Alkaline phosphatase(ALP)and alizarin red S staining were used to determine the ALP activity and mineralization after osteogenic induction and culture. Real-time fluorescence quantitative PCR and Western blot were used to detect mRNA and protein expressions of osteoblast related genes. Results:Alogliptin intervention improved the biochemical indexes of bone anabolism and protected against bone microstructure deterioration to alleviate bone loss in OVX mice. Alogliptin stimulated osteoblast differentiation and elevated expression levels of Runt-related transcription factor 2(Runx2), ALP, osteocalcin, and osterix in in vitro experiments. Conclusion:Alogliptin can alleviate bone loss in OVX mice.

Objective:To observe the main clinical features and outcomes of type 2 diabetes mellitus patients after infection with COVID-19 and to compare them with those without diabetes mellitus.Methods:A single-center retrospective observational study was conducted in 88 in-patients who were diagnosed as COVID-19 from January 1 to February 26, 2020. They were divided into diabetic group and non-diabetic group, with 44 patients in each group. Patients′ medical history, laboratory examination, in-hospital treatment plan, and disease outcome were collected and compared.Results:The clinical symptoms of diabetic patients were varied, mainly fever(75.0%), cough(75.0%), fatigue(52.3%), and so on. The systolic blood pressure(SBP)was higher [131.50(120.00, 140.75) vs 125.00(120.00, 131.75)mmHg, 1 mmHg=0.133 kPa, P=0.021] and the oxygenation was lower [96.00%(94.25%, 97.00%) vs 97.00%(95.00%, 98.00%), P=0.038] at admission compared with the non-diabetic group. Hypertension and chronic kidney disease were more common in diabetic group. Fasting blood glucose [7.64(6.12, 15.43) vs 5.62(5.25, 6.50)mmol/L, P<0.01], interleukin-6(IL-6)[19.85(6.50, 43.38) vs 10.80(3.03, 20.90)pg/ml, P=0.046] in diabetic group were significantly higher than those in non-diabetic group. Secondary infection(27.3% vs 9.1%, P=0.027), ARDS(22.7% vs 4.5%, P=0.013)and shock(4.5% vs 0%, P<0.01)were more likely to occur in the diabetic group. More patients were treated with mechanical ventilation in the diabetic group(20.5% vs 4.5%, P=0.024). The diabetes group was more likely to progress to critical type(20.5% vs 4.5%, P=0.024), and the time to progress to critical state was shorter [3(1.75, 5.25) vs 6(3.00, 12.00)d, P=0.019). The duration of severe symptoms was longer in the diabetic group [26.5(15.00, 31.50) vs 9(8.00, 13.00)d, P=0.026]. Conclusion:The clinical symptoms of type 2 diabetes patients with COVID-19 are diverse. They are often combined with diseases such as hypertension and chronic kidney disease. The inflammatory reaction is more obvious and has more COVID-19 related complications and is more likely to progress into a persistent severe condition in a short time.

Objective To investigate the effect of myeloid-derived growth factor ( MYDGF) on the secretion of glucagon-like peptide 1 ( GLP-1) in type 2 diabetic mice and its mechanism. Methods A type 2 diabetes model was established by injecting streptozotocin into C57BL/6J wild type ( WT) mice and MYDGF knockout ( KO) mice, which were divided into diabetic group ( WT-D, KO-D) and non-diabetic group ( WT-ND, KO-ND) . Six weeks later, the relevant indicators were detected. Next, those mice were divided into wild-type diabetes group (WT-GFP), wild-type diabetes MYDGF intervention group (WT-MYDGF), knockout type diabetes group (KO-GFP), and knockout type MYDGF intervention group ( KO-MYDG ) according to whether or not the AAV-MYDGF intervention was performed. The wild-type non-diabetic mice were used as a blank control group to observe the effects of MYDGF on biochemical indexes, GLP-1 secretion, and mitogen-activated protein kinase kinase ( MEK)/extracellular regulated protein kinases ( ERK) signal pathway in mice. Results After 6 weeks of intervention, there was no significant difference in the glucose and lipid metabolism indexes between WT-ND and KO-ND groups ( P>0.05) . Compared with WT-D group, fasting plasma glucose (FPG), HbA1C, and blood lipid levels in KO-D group were increased, while gcg, pc3 mRNA, and GLP-1 secretion levels were decreased (all P<0.05). Compared with the WT-GFP group, FPG, HbA1C , and blood lipid levels were decreased in WT-MYDGF group, while gcg and pc3 mRNA, and GLP-1 secretion levels were increased (all P<0.05). KO group revealed a result similar to that in WT group after MYDGF intervention. Western blotting showed that the phosphorylation level of ERK1/2 was lowered in KO diabetic mice compared with WT diabetic mice, which was enhanced in WT and KO mice after MYDGF intervention. Conclusions MYDGF promotes the secretion of GLP-1 by activating MEK/ERK signaling pathway, thereby delaying the development of diabetes.

Objective To investigate the effects of growth differentiation factor 11 ( GDF11 ) on β-cell function in db/db mice and its possible mechanism. Methods Twenty eight-week-old male db/db mice were randomizedtoi.p. administration of GDF11(0.3mg·kg-1·day-1)or equivalent PBS(n=10)for 6 weeks.10age-matched male db/m were used as normal control, received equivalent PBS injection for 6 weeks. Blood glucose levels, body weights and food intake were monitored weekly. IPGTT and glucose-stimulated insulin secretion ( GSIS) were analyzed. After 6 weeks of intervention, serum HbA1C , TG, TC, and FFA were measured respectively. The concentrations of hormones in serum and pancreas were evaluated. The mRNA expression of Pdx-1, MafA, Nkx6. 1, and insulin2 were determined by RT-PCR. The expression of phosphorylated Smd2 (P-Smad2), Smad2 in islet were examined by western blot. Results Compared with NC group, GDF11 administration decreased FBG, HbA1C , modified lipid profiles. GDF11 improved glucose tolerance and augmented GSIS. Moreover, GDF11 increased serum insulin and pancreatic insulin content, while decreased serum glucagon concentration. The expression of Pdx-1, MafA, Nkx6. 1, and Insulin2 were significantly increased in GDF11 group. GDF11 elevated the expression of P-Smad2 in islets. Conclusion s GDF11 may preserve β-cell function and facilitate the secretion and production of insulin. Diminishing the metabolic abnormalities, alleviating the secretion of glucagon, as well as maintaining the key transcript factor activation may contribute to the amelioration of β-cell function after GDF11 administration. Smad2 pathway may be related to the protective effects of GDF11.

Objective To assess the association of subclinical thyroid dysfunction with fractures. Methods Medline, Embase, Pubmed, Cochrane Library, CBM, CNKI, Wan Fang, and VIP databases were systematically searched from January 1990 to August 2015 to identify prospective cohort studies which have studied the risk of fracture in patients with subclinical thyroid dysfunction. The relative risks ( RR) of cohort studies were pooled respectively, depending on the result of heterogeneity test among the individual studies search. The Stata (version 13. 0) software was used for meta-analysis. Results Nine prospective cohort studies including 292460 participants were identified as eligible for the meta-analysis. RR of subclinical hyperthyroidism for fracture was 1. 39(95%CI 1. 24-1. 55);for hip fracture, RR was 1. 24(95%CI 1. 10-1. 40);for nonspine fracture, RR was 1. 32 (95%CI 1. 09-1. 60). Different gender for subclinical hyperthyroid was associated with higher fracture rates:for females, RR was 1. 15(95%CI 1. 04-1. 27); for males, RR was 1. 31 (95% CI 1. 08-1. 59). The incidence of fracture in patients with subclinical hyperthyroidism was higher during the follow-up. For subclinical hypothyroidism, the RR was 1. 21(95% CI 1. 03-1. 42). Subgroup analysis indicated that there were significant differences between endogenous/exogenous subclinical hyperthyroidism and euthyroid, but no differences between endogenous/exogenous subclinical hypothyroidism and euthyroid were found. Conclusion Subclinical hyperthyroidism is associated with an increased risk of fracture in the population, especially hip fracture and nonspine fracture. During the course of subclinical hyperthyroidism, the incidences of fracture should be noticed both in females and males. However, there is no evidence which could prove a definite association between subclinical hypothyroidism and the risk of fracture.

Objective To investigate the effects and possible mechanism of growth differentiation factor 11 (GDF11)on angiogenesis in diabetic hindlimb ischemia. Methods Sixty SD rats were used in this study. Diabetes was induced by intraperitoneal injection of streptozotocin. 3 days after streptozotocin administration, 40 rats with plasma glucose concentration≥16.7 mmol/L were selected in the subsequent experiments. 12 weeks after diabetes was induced,the left femoral artery and all the sides branches were dissected free and excised. After resection of the left femoral artery,rats were randomized to four groups:PBS group(n=10),GDF11 group(n=10),IgG Ab group (n=10),or GDF11 Ab group(n=10). After 0,7,and 14 days,the serial blood flows were measured by a Laser Doppler perfusion image(LDPI)analyzer. To detect capillary endothelial cell,the sections of muscles were reacted with anti-CD31 monoclonal antibodies,and subsequently reacted with Cy3-conjugated anti-rabbit IgG antibody. The expression levels of HIF1α and VEGF were detected by western blotting. Results In GDF11 group significantly increased the blood perfusion and capillary density of ischemia hindlimb of the diabetic rats were found,which was correlated to an increased level of HIF1α and VEGF. In contrast, GDF11 Ab could lead to the opposite effects. Conclusion GDF11 treatment promotes the recovery of diabetic hindlimb ischemia, which may be related to the improvement of expression of HIF1 alpha and VEGF.

Objective To study the effect and mechanisms of berberine (BBR) on the proliferation of papillary thyroid cancer K1 cells induced by high glucose.Methods K1 cells were cultured under 5.5 mmol/L or 25 mmol/L glucose condition with or without different concentration of BBR (0,10,40 and 80 μmol/L) for 24 hours.The proliferations of K1 cells in each condition were detected by MTT.Western blot was used to measure the expression of nuclear factor erythroid 2-related factor 2 (Nrt2),phosphoinositide 3-kinase (PI3K),protein kinase B (Akt) and phosphorylated-Akt (p-Akt).The distribution pattern of Nrf2 in K1 cells was determined using immunofluorescent staining.Results Compared with 5.5 mmol/L condition,the proliferation rate [(126.64 ± 5.41) ％ vs (87.31 ± 3.67) ％],expression levels of PI3K (0.425 ±0.019 vs 0.272 ±0.039),p-Akt/Akt (0.446 ±0.021 vs 0.168 ±0.035) and Nrf2 (0.597 ± 0.014 vs 0.308 ± 0.026),and Nrf2 distribution (93.0％ vs 23.1％) in nuclear of K 1 cells under 25 mmol/L condition were significantly elevated,respectively (all P ＜0.01).Addition of BBR in 25 mmol/L condition dose dependently (10,40,80 μmol/L) lowered the proliferation rate of K1 cells [(111.76 ± 4.10)％,(70.03 ±2.18)％,(32.41 ±3.76)％ vs (126.64 ±5.41)％,all P＜0.05],and suppressed the expression of PI3K,p-Akt/Akt,Nrf2,and Nrf2 nuclear distribution (P ＜ 0.05).Conclusions BBR dose dependently inhibited the proliferation of high glucose-induced K1 cells.This effect was associated with the suppression on of PI3K/Akt signaling activation,Nrf2 expression and its nuclear translocation.