Objective:To analyze the clinical phenotype and genetic characteristics of a Chinese pedigree affected with Hereditary antithrombin deficiency.Methods:A pedigree diagnosed at the the Second Affiliated Hospital of Wenzhou Medical University, Yuying Children’s Hospital in June, 2020 was selected as the study subject. Plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), and thrombin time (TT) of the probands and their pedigree members were determined using a STA-R automatic coagulation analyzer. Antithrombin activity (AT: A) and antithrombin antigen (AT: Ag) in plasma were determined with chromogenic substrate and immunonephelometry assays. All exons and flanking sequences of the anticoagulant protein gene SERPINC1 were amplified by PCR and subjected to Sanger sequencing. Candidate variants were verified with bioinformatic tools (PolyPhen-2, SIFT, Mutation Taster and PYMOL) to explore their effect on the function and structural conformation of the protein. Results:The probands (Ⅱ 2, Ⅱ 10), their brother (Ⅱ 5) and sons (Ⅲ 1, Ⅲ 8) had shown normal PT, APTT, FIB, and TT, but significantly decreased AT: A and AT: Ag, with their levels being 34%, 57%, 56%, 48%, 53% and 13.51 mg/dL, 13.44 mg/dL, 18.39 mg/dL, 17.36 mg/dL, 17.71 mg/dL, respectively. The remaining pedigree members had normal values. Sanger sequencing revealed that the probands and all affected pedigree members had harbored a heterozygous c. 851T>C (p.Met284Thr) missense variant in exon 5 of the SERPINC1 gene. Bioinformatic analysis and simulation suggested that the variant has resulted in alteration of hydrogen bonds at the c. 851 position, which may affect the structure of the protein. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PS1+ PM1+ PM5+ PP1+ PP4). Conclusion:The probands and other affected members were all diagnosed with type I hereditary AT deficiency, for which the c. 851 T>C (p.Met284Thr) variant of the SERPINC1 gene may be accountable.

As a new hypoglycemic drug,Dapagliflozin has attracted much attention because of its unique hypoglycemic mechanism. It has been used in many studies on type 2 diabetes mellitus,but the application of type 1 diabetes mellitus(T1DM)in the eastern population is rare. This article uses Dapagliflozin through a case of obese T1DM to provide new ideas for the treatment of T1DM.

Objective To investigate the dynamic mechanical responses and damage characteristics of peri-implant bone structures subjected to impact load.Methods A finite element model of the peri-implant bone microstructure was established,and an initial velocity was applied to the rigid body to simulate the impact load.A stress failure criterion was employed and a user-material subroutine was developed to assess failure.Subsequently,bone damage after the impact load was analyzed according to the material subroutine.Results After the impact load,the stress on the cortical bone increased rapidly,reaching a peak value(16.01 MPa)immediately.In contrast,the stress on the trabecular bone at the bottom of the implant reached its peak value(5.85 MPa)at 0.1 μs.The impact load resulted in stress waves that propagated and diffused within the bone structure,causing changes in the bone structure damage over time.The generated impact energy could be absorbed and dissipated by the trabecular bone through deformation.The deformed trabecular bone experienced damage and failure upon reaching the yield limit,whereas the cortical bone did not experience damage or failure under an impact load.Conclusions Structural changes in the trabecular bone should be considered in patients with impact damage.The numerical model established in this study can effectively predict bone impact damage by combining the structural mechanical properties and geometric characteristics of the bones.These findings can serve as a reference for assessing bone damage and post-damage treatment in patients subjected to impact loads in clinical practice.

Objective:To explore the effects and mechanism of Bushen Tongluo Prescription in mechanical injury of rat endometrium.Method:A total of 60 female SD rats were divided into the blank group, model group, estradiol valerate group, and Bushen Tongluo Prescription group according to the random number table method, with 15 rats in each group. Except for the blank group, the other three groups were used to establish a rat model of endometrial injury using mechanical injury method. After modeling, Bushen Tongluo Prescription group was orally administered with Bushen Tongluo Prescription decoction at a dosage of 2.1 g/kg, the estradiol group was orally administered with estradiol valerate at a dosage of 0.4 mg/kg, and the blank group and model group were orally administered with 0.5% CMC at an equal dosage, once a day, for a total of 8 days. Samples were taken on the 1st, 4th, and 8th day after gastric lavage. The thickness of the endometrium and glands were observed using HE staining, the degree of uterine tissue fibrosis was observed using Masson staining, and VEGF and TGF-β protein and mRNA expressions in uterine tissue were detected using Western blot and fluorescence quantitative PCR.Results:Compared with the Bushen Tongluo Prescription on the first day, the thickness of the endometrium and the number of glands increased on the eighth day ( P<0.05); compared with the model group, the number of glands in the Bushen Tongluo Prescription group increased on the 4th day of administration ( P<0.05), while the fibrotic area of the endometrium decreased on the 8th day of administration ( P<0.05); compared with the model group, on the 8th day of administration, the expression of VEGF protein and TGF-β1 in the Bushen Tongluo Prescription group increased ( P<0.05), and protein expression decreased ( P<0.05); compared with the model group, the group treated with Bushen Tongluo Prescription had TGFβ1 mRNA level on the first day increased ( P<0.05), while the level of TGF-β1mRNA decreased on the 8th day ( P<0.05). Conclusion:Bushen Tongluo Prescription can increase endometrial thickness and receptivity, effectively improve damaged endometrium, anti fibrotic and prevent endometrial adhesion by up-regulating VEGF protein expression and down-regulating TGF-β1 protein and mRNA expression.

Objective:To explore the effects of game addiction disorders on brain cognitive control functions based on near-infrared spectroscopy.Methods:Thirteen subjects were screened according to the Online Game Addiction (OGA) Scale. The experimental paradigm was the stop-signal task. The relative concentration levels of oxyhemoglobin (HbO 2) and deoxyhemoglobin (Hb) in the prefrontal region of the brain during cognitive activity were collected using near-infrared spectroscopy to assess the cognitive control function of the subjects. Results:The game-addicted patients had lower keystroke accuracy in the stop-signal task than healthy subjects, and the difference was statistically significant ( P<0.05). Compared to healthy subjects, game-addicted patients had less activation in prefrontal areas and showed uncontrolled behavior and brain activity. Conclusions:Game addiction disorders impair brain cognitive control, which in turn triggers a weakening of cognitive control. The results of this study provide a reference for the prevention and treatment of game addiction.

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.

Objective:To investigate prognostic factors for acral lentiginous melanoma (ALM) , and to construct a nomogram to verify the predictive value of these factors.Methods:Clinical data on 1 573 patients with ALM were collected from the Surveillance, Epidemiology, and End Results (SEER) database of National Cancer Institute in United States between 2004 and 2015. Data about patients′ age, gender, ulcer status, SEER staging, surgical protocols, T-, N- and M-staging, overall survival rates and disease-specific survival rates were extracted. Chi-square test was used to analyze the correlation of clinical characteristics with overall survival rates and melanoma-specific survival rates, and univariate and multivariate Cox proportional hazards regression models were used to investigate prognostic factors and establish predictive models.Results:Among the 1 537 patients with ALM, 714 were males, 823 were females, 818 were under 64 years of age, and 1 363 were Caucasian. Skin lesions occurred on the lower limbs and buttocks in 1 205 cases, and 974 cases had ulcers; according to the SEER staging, non-spread localized skin lesions were observed in 1 048 cases. There were significant differences in the mortality rate among patients of different ages at diagnosis, different gender, with different ulcer status, surgical status, and at different SEER stages, T-stages, N-stages and M-stages (all P < 0.001) . Univariate and multivariate Cox regression analysis showed that age ≥ 65 years, male, ulcers and distant lymph node metastasis in the SEER staging were associated with increased risk of death in the patients (all P < 0.05) , and the mortality rate was significantly higher in the patients with T2-, T3- or T4-stage ALM than in those with T1-stage ALM (all P < 0.05) , and higher in the patients with N1-, N2- and N3-stage ALM than in those with N0-stage ALM (all P < 0.05) . Conclusion:Age, gender, ulcer status, SEER stage, T-stage and N-stage are independent prognostic factors for overall survival rates and disease-specific survival rates of ALM.

Objective:To analyze the phenotype and genotype of two Chinese family with inherited dysfibrinogenemia and the molecular pathogenic mechanism.Methods:In the probands and their family members, coagulation routine, fibrinogen activity(Fg∶A) and fibrinogen antigen(Fg∶Ag) were detected . To find the mutation and exclude single nucleotide polymorphisms, all the exons and exons-intron boundaries of fibrinogen genes ( FGA, FGB and FGG) were amplified by Ploymerase Chain Reaction (PCR), then sequenced. Bioinformatics prediction softwares were used to predict and score the change of function caused by the variant. PyMol were used to analyze the structure of protein caused by the variant. Clustal X software was used to analyze the conservation of the mutant amino acids. Results:The thrombin time(TT) of the two was slightly prolonged and could not be corrected by protamine sulfate, and the fibrinogen activity was significantly reduced (1.25 g/L and 1.17 g/L), but the fibrinogen antigen content was normal, respectively (3.50 g /L and 3.81 g/L). Genetic analysis showed that both probands were heterozygous missense variants( FGB exon 7 c. 1115 T>A (p.Val372Glu)), both of which originated from the paternal line. The prediction results of the four bioinformatics softwares indicate that this variant could be disease causing. Clustal X software showed that Val372 is highly conserved among homologous species. Based on the guidelines of the American College of Medical Genetics and Genomics, c. 1115 T>A was predicted to be likely pathgenic(PM2+ PP1+ PP2+ PP3+ PP4). PyMol shouwed p. Val372Glu variant changes the secondary structure and three-dimensional structure of fibrinogen protein were changed caused by p. Val372Glu variant. Conclusion:Inherited dysfibrinogenemia of the probands maybe caused by variant of FGB c. 1115 T>A(p.Val372Glu), and the variant was firstly reported.

Objective:To investigate the intervention effect of topical shikonin on an imiquimod-induced psoriasis-like mouse model and its effect on expression of CCAAT enhancer binding protein δ (CEBPD) .Methods:Twenty specific pathogen-free BALB/c male mice were randomly and equally divided into model group, shikonin 1 group, shikonin 2 group and blank control group by using simple random sampling. Mice in the model group, shikonin 1 group and shikonin 2 group were topically treated with 50 mg of 5% imiquimod cream every day on the shaved back to establish the psoriasis-like mouse model. After 6-hour treatment, mice in the shikonin 1 group and shikonin 2 group were treated with 0.5 ml of shikonin at concentrations of 0.576 and 5.76 g/L respectively in the modeling area for 8 consecutive days; the blank control group received no treatment. Changes in the skin lesions of these mice were observed by naked eyes every day, and evaluated by using psoriasis area severity index (PASI) ; after 8-day treatment, the mice were sacrificed by cervical dislocation, the dorsal skin tissues were resected, and immunohistochemical study and Western blot analysis were performed to determine the expression of CEBPD in the mouse epidermis. Statistical analysis was carried out with SPSS 16.0 software by using one-way analysis of variance for comparisons of observation indices among different groups, as well as least significant difference- t test for multiple comparisons. Results:On day 8, the mice in the model group presented with obvious erythema, scales, and infiltrative and thickened skin lesions; compared with the model group, the skin lesions were markedly improved in the shikonin 1 group and shikonin 2 group, and the improvement was more obvious in the shikonin 2 group. On day 8, the PASI score significantly differed among the blank control group, model group, shikonin 1 group and shikonin 2 group (0, 11.0±1.22, 8.6±0.55, 5.8±1.30 points, respectively; F=128.21, P<0.01) , and there were significant differences between any two groups (all P < 0.01) . Immunohistochemical study showed a significant difference in the expression of CEBPD ( A value) among the model group, shikonin 1 group, shikonin 2 group and blank control group (0.072±0.026, 0.177±0.036, 0.290±0.062, 0.407±0.051, respectively; F=48.895, P < 0.01) , and there were also significant differences between any two groups (all P < 0.01) . Western blot analysis showed that the CEBPD expression in the mouse epidermis was highest in the blank control group, followed in descending order by the shikonin 2 group, shikonin 1 group and model group, and significantly differed among the above 4 groups ( F=10.237, P<0.05) ; moreover, there were significant differences in the CEBPD expression between the model group and blank control group, as well as between the shikonin 1 group and blank control group (both P<0.05) , while no significant difference was observed between the shikonin 2 group and the blank control group ( P > 0.05) . Conclusion:Topical shikonin could effectively interfere with the development of imiquimod-induced psoriasis-like mouse model; CEBPD expression decreased in the psoriasis-like mouse model, and could be markedly upregulated by topical application of shikonin.

OBJECTIVE: To analyze the phenotype and genotype of a patient affected with inherited antithrombin deficiency. METHODS: All exons and exon-intron boundaries of the AT genes were subjected to PCR amplification and Sanger sequencing. The influence of variants on the disease was predicted using bioinformatic software (MutationTaster). RESULTS: The results of all coagulation tests were normal, though the antithrombin activity and antigen content of the proband and his father have decreased significantly (34%, 48% and 12.97 mg/dL, 15.60 mg/dL, respectively). His mother was normal. Genetic analysis revealed that the proband and his father both carried a heterozygous g.2736dupT variant of the AT gene. Bioinformatic analysis suggested that the variant may be pathogenic. CONCLUSION The proband and his father both had type I hereditary antithrombin deficiency caused by a g.2736dupT variant of the AT gene. The variant was unreported previously.
Antithrombin III/genetics* ; Antithrombin III Deficiency/genetics* ; DNA Mutational Analysis ; Genetic Testing ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree