Objective To study the effects of metformin on proliferation and apoptosis of oral squamous cells so as to create a new path for the treatment of oral carcinoma.Methods CAL27 cells were treated with different dosage of metformin(5,10,20 mmol/L)for 24,48, and 72 hours.The number of apoptosis was detected by flow cytometry and TUNEL.The autophagic vacuole was detected by immunofluores-cence.The expression of hallmark of apoptosis and autophagy was detected by Western blot.Results Metformin could induce apoptosis in CAL27 cells.The CL-PARP and Bcl-2 expression significantly increased.In line with the apoptosis,metformin can trigger autophagy in CAL27 cells.The expression of LC3,Beclin-1 and GFP-LC3 positive autophagosomes were increased by metformin.Metformin inhibit the expression of STAT3 and mTOR signaling pathways at the same time might be the possible reasons of the autophagy.And inhibited autophagy could en-hance metformin induced Caspase-3 activity in CAL27 cells.Conclusion Inactivation of STAT3 and mTOR pathway contributes to metform-in-induced autophagy.Inhibited autophagy could enhance metformin induced apoptosis in oral squamous cell carcinoma.

Objective To evaluate the clinical value of microvascular anastomotic devices in free anterolateral thigh flap in the recon-struction of oral carcinoma defects.Methods From September 2013 to September 2014,52 patients with oral cancer were treated by func-tional neck cleaning,primary tumors resection,and free anterolateral thigh flap in the reconstruction.Of which 27 patients received free of femoral anterolateral flap to repair veins by using microsurgical line end to end anastomosis.And 25 pieces of microvascular anastomotic ad-vices were applied in 25 patients with oral carcinoma defects.The length of anastomosis time,flap survival rate,and complications were recor-ded.Results In 52 patients,microvascular anastomotic devices were applied in 25 veins with shorter anastomosis time,(6.3 ±1.9)min vs. (12.3 ±1.4)min,and the difference was significant(P =0.001).The flap survival rate and vascular anastomosis patency rate was 100%.No intra-and post-operative complications such as blood leak,stapling,excessive tension and tear,thrombosis in flap vein crisis associated with microvascular anastomotic devices were observed.Twenty-seven patients with 40 root vein received manual microsurgical anastomosis,2 of them showed vein crisis.Conclusion The microvascular anastomotic devices used in repairing the soft tissue defects by anterolateral thigh flap with venous anastomosis for patients of oral cancer after surgery has the advantage of higher quality,shorter time and less complications.

Objective To compare the results of GnRHa stimulation test and GnRH stimulation test in girls with Idiopathic central precocious puberty.Methods The girls aged 6-10 who were diagnosed with early breast development (＜ 8 years old) from January 2016 to June 2018 were randomly divided into GnRHa stimulation test group and GnRH stimulation test group according to clinical manifestations,auxiliary examinations and follow-up.The difference in the results of the stimulation test between the two groups was compared.Results A total of 108 girls were enrolled in this study,and 40 (37％) of the GnRHa stimulation tests were performed.Among them,25 (63％) patients with idiopathic central precocious puberty had the confirmed diagnosis,and the GnRH stimulation test was performed.Of the 68 (63％) patients,30 (44％) had idiopathic central precocious puberty.The peaks of LH in the GnRHa challenge test group and the GnRH challenge test groupwere 11.33 (6.81,15.79) and 7.89 (5.35,14.21),and the FSH peaks were 15.68 (10.18,20.06) and 17.26 (13.34,21.42),showing no significant differences (U =1078.50,P =0.07;U =1617.50,P=0.10).Thepeak values of LH/FSH were0.86 (0.37,1.17)and0.52 (0.31,0.83),respectively,and there was a statistical difference (U =953.00,P =0.01).GnRH challenge test showed the ICPP sensitivity of 90.91％ (81.10％-100.72％),specificity of 94.29％ (86.60％-101.98％),total coincidence rate of 92.65％ (86.44 ％-98.85％),Yoden index of 0.85 (0.72％-0.98％).Sensitivity of the GnRHa challenge test showed the ICPP diagnosis rate of 95.24％ (86.13％-104.35％),specificity of 73.68％ (53.88％-93.48％),total coincidence rate of 85.00％ (73.93％-96.07％),Yoden index of 0.69 (0.47-0.91).Conclusion For the diagnosis of idiopathic central precocious puberty in girls,the GnRHa challenge test is more sensitive than the GnRH challenge test,and the specificity and the Yoden index are both low.Therefore,routine use of GnRHa stimulation test as the replacement of GnRH stimulation test is not recommended.

OBJECTIVE: To summarize clinical manifestations, inheritance pattern and mutations of NR0B1 gene in 7 children with X-linked adrenal dysplasia congenita (XL-AHC). METHODS: Clinical data of the 7 children was collected. Next-generation sequencing was carried out to detect potential mutations in the coding regions of adrenal gland-related genes. Suspected mutations were verified with Sanger sequencing. RESULTS: In all of the children, the initial symptom was adrenocortical insufficiency. Five cases had neonatal onset, while the remaining two developed it at the age of 2. Three cases (42.9%) had a short stature and 1 showed growth retardation (14.3%). Of the 7 cases, 6 (85.7%) had mutations occurring in exon 1, and 1 (14.3%) had it occurring in exon 2. Four cases (57.1%) were frameshift mutations, 2 cases (28.6%) were nonsense mutations and 1 case (14.3%) was missense mutation. Two mutations were known to be pathogenic, and 5 had not been reported previously. Maternal inheritance was found in 6 cases. Three children had a maternal uncle died of unexplained causes. The mothers of 2 children had a history of spontaneous abortions. One child had a brother died of unexplained reason. CONCLUSION Male children with primary adrenal insufficiency should be routinely checked for NR0B1 mutations, especially those with a family history. mutations of NR0B1 gene occur mostly in exon 1, with frameshift mutations being the most common type. The development of all patients with XL-AHC should be closely monitored during follow-up.
Adrenal Insufficiency ; Child ; DAX-1 Orphan Nuclear Receptor ; DNA Mutational Analysis ; Genes, X-Linked ; Humans ; Hypoadrenocorticism, Familial ; Male ; Mutation

OBJECTIVETo analyze clinical characteristics, genetic mutation and therapeutic effect of seven patients diagnosed with congenital hyperinsulinism(CHI).

METHODSClinical data for the patients was retrospectively analyzed.

RESULTSAll patients presented with hyperinsulinism(serum insulin:2.0-58.4 mU/L),even after hypoglycemia (blood glucose: 0.7-2.39 mmol/L) has developed. Mutations were identified in 4 patients (57.1%), which included a heterozygous c.262C to T(p.R88C) mutation in exon 4 of the UCP2 gene, a heterozygous c.1495C to A(p.G499C) mutation in exon 12 of the GLUD1 gene, a heterozygous c.1493C to T(p.S498L) mutation in exon 1 of the GLUD1 gene, and a heterozygous c.4432G to A(p.G1478R) mutation in exon 37 of the ABCC8 gene. The patient carrying a maternally inherited ABCC8 mutation was treated with cornstarch and had his blood glucose kept normal. All other patients responded well to diazoxide.

CONCLUSIONA genetic diagnosis was attained for 51.7% of patients in this study. Mild CHI patients can have their blood glucose controlled by giving cornstarch. Diazoxide is safe and effective for most CHI patients.