Objective Celloidin sections of temporal bone were used to develop a serial stereo anatomical atlas of temporal bone by computer aided three dimensional(3-D) reconstruction and stereology. Methods Fifty sets of serial celloidin sections of temporal bone with reference points were prepared and 3-D morphology of structures in temporal bone was restored systemically by the technique of computer aided 3-D reconstruction. A system of stereo anatomical atlas of temporal bone with stereoscope was established. Results Totally 48 cases of reconstruction were accomplished for bony and membranous labyrinth, ossicles, tendons, facial nerve and its canal, round window membrane and niche, posterior ampullary nerve, endolymphatic sac, cochlear aqueduct, VII and VIIIth cranial nerve. The stereo picture pairs of these structures with the best representation were selected as the stereo anatomical atlas of temporal bone which showed the fine spatial morphology and relationship of the above structures. The stereo anatomical atlas was applied to guide ear surgery and to assist anatomic training of temporal bone. Conclusion The stereo anatomic atlas of temporal bone is an innovative powerful anatomic tool and has tremendous application futures in otology, or even in the whole medical science.

Objective To assess the value of four different techniques of detecting the Mycobacterium tuberculosis (MTB) in bronchoalveolar lavage fluid (BALF) in the diagnosis of tracheobronchial tuberculosis. Methods A total of 98 patients diagnosed as tracheobronchial tuberculosis were selected from May 1,2013 to June 30,2016. The clinical data was analyzed retrospectively,and the positive rates of MTB of the 960 cultrue, the direct smears , the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were compared. Results The positive rates of the 960 cultrue,the direct smears,the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were 20.4%(20/98),15.3%(15/98),70.4%(69/98) and 74.5%(73/98),respectively. Among the four techniques ,the positive rates of the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were significantly higher than those of the 960 cultrue and the direct smears(P<0.05,respectively). However,no significant difference was found between the modified Ziehl?Neelsen stain method and the Xpert MTB/RIF assay (P > 0.05). Conclusions The modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay for detecting the MTB in BALF have high clinical value in the diagnosis of tracheobronchial tuberculosis.

To analyze the mutations of F12 genein one pedigree with congenital factor FXII (FXII) deficiency , and investigatethe molecular mechanisms of FXII deficiency . Methods Activated partial thromboplastin time(APTT),Prothrombin time(PT), FXII activity(FXII:C), FXII antigen(FXII:Ag) and other coagulant parameters were tested in the proband and his family members .5'and 3'UTR,all exons and their exon-intron boundaries of F12 gene were analyzed by direct sequencing .The detected mutations were confirmed by reverse sequencing .100 healthy persons were as normal controls .Results The proband showed a markedly prolonged APTT (106.4s), the FXII:C and FXII:Ag were 2.0% and 1.0%, respectively .Hissecond daughter and granddaughter had slightly prolonged APTT , and other family members are normal.The FXII:C and FXII:Ag of family members were also decreased ( his son, 23.0% and 21. 0%;his elder daughter , 23.0%and 23.0%;his second daughter ,24.0%and 23.0%;hisgranddaughter , 23.0%and 23.0%).The phenotype of all members is consistent with cross -reactive material negative . Nucleotide sequencing analysis showed that the proband had missense mutations in the F 12 gene, including one homozygous mutationc.1556T >G ( p.Leu519Arg) and a commonly reported single nucleotide polymorphism site within the promoter region of the F 12 gene (46T/T) .Sequencing results from the proband 'children demonstrate them as carriers of a heterozygous missense mutation .The proband 's wife is normal and with 46C/C in the promoter region .Conclusion The c.1556T>G in exon 13 is a novel mutation .This mutation affects FXIIcatalytic function , associated with a reduced level of FXII .

OBJECTIVE: To explore the changes of serum interleukin-6 (IL-6) level in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and OSAHS associated type 2 diabetes mellitus (T2DM) and their significance. METHOD: All observed subjects were divided into 3 groups: 20 cases of normal subjects, 35 cases of simple OSAHS patients, 48 cases of OSAHS associated T2DM patients, IL-6 concentrations of serum were measured by the enzyme-linked immunosorbent. RESULT: IL-6 level was higher in the group of OSAHS with T2DM than the group of OSAHS and the healthy control group (P < 0.05); IL-6 level was higher in the simple OSAHS group than the healthy controls. CONCLUSION IL-6 and other inflammatory factors may involved pathological physiological process in OSAHS patients sugar metabolic abnormalities; and is associated with the development of OSAHS associated with type 2 diabetes.
Case-Control Studies ; Diabetes Mellitus, Type 2 ; blood ; complications ; Humans ; Interleukin-6 ; blood ; Sleep Apnea, Obstructive ; blood ; complications

OBJECTIVE: To investigate the significance of tumor necrosis factor-alpha (TNF-α) and plasma endothelium (ET) in pathophysiologic process of patients with obstructive sleep apnea hypopnea syndrome(OSAHS) with type 2 diabetes mellitus (T2DM). METHOD: All observed subjects were divided into 4 groups. A number of 80 patients with OSAHS, 65 cases of OSAHS with T2DM patients, 20 patients with T2DM, and 32 cases of healthy control group were observed in this study. The serum levels of TNF-α and ET were detected by double antibody sandwich ELISA, the content of TNF-α and ET were compared between OSAHS group and OSAHS + T2DM group. It were also compared before and after treatment of CPAP or surgery. RESULT: TNF-α level is higher in OSAHS+T2DM group than that in the OSAHS group, T2DM group and the control group (P < 0.05); TNF-α level in OSAHS group and the T2DM group are higher than that in the control group (P < 0.05), but there was no difference in TNF-α level between the OSAHS group and the T2DM group. There was also no difference in ET level of the four groups. There were significant differences of TNF-α before and after treatment of CPAP or surgery in OSAHS group and OSAHS+T2DM group (P < 0.05). But there was no significant differences of ET before and after treatment of CPAP or surgery in the OSAHS+T2DM group (P > 0.05). CONCLUSION TNF-α may be involved in the development of OSAHS and T2DM, while ET may have little effect on the occurrence and development of OSAHS and T2DM.
Antibodies ; Diabetes Mellitus, Type 2 ; blood ; complications ; Endothelins ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Sleep Apnea, Obstructive ; blood ; complications ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVE: To explore the risk factors of type 2 diabetes mellitus (T2DM) of Dong nationality in Western Hunan, and to provide a scientific basis for the prevention and treatment of T2DM in the district. METHODS: In the case-control study, the subjects were divided into a T2DM group, an impaired glucose regulation (IGR) group and a normal glucose tolerance (NGT) group through oral glucose tolerance test (OGTT). A questionnaire survey was conducted, and physical measurements and the detection of blood glucose, blood lipids and serum insulin were done. RESULTS: Univariate analysis found significant difference in age, fasting insulin (FINS), HOMA insulin resistance index (HOMA-IRI), HOMA β-cell function index (HOMA-βC), trigalloyl glycerol (TG), high density lipoprotein cholesterol (HDL-C), systolic blood pressure, and diastolic blood pressure in the glucose metabolism among different groups (P<0.05). Body mass index (BMI), waist/hip ratio (WHR), family history of diabetes, number of physical activities per week, dinner taste, fat and protein-rich foods, fresh vegetables and fruits intake were associated with T2DM. Multifactor non-conditional ordinal logistic regression indicated that age was the risk factor of T2DM and IGR. Compared with the group whose age was≤50, the odds ratios of the groups of 5070 were 1.85, 2.83 and 2.64 respectively, P<0.05. The risk of suffering from diabetes of the overweighted or obese people was 2.13 times that of a normal BMI group, P<0.01. The other influencing factors included WHR (OR=2.06), family history of diabetes (OR=11.36), and fat and protein-rich foods (OR=1.90). CONCLUSION The main influencing factors of T2DM of Dong nationality in Western Hunan include age, BMI, WHR, family history of diabetes, fat and protein-rich foods. We must strengthen the health eduation of T2DM of Dong nationality in Western Hunan to reduce the risk of T2DM.
Blood Glucose ; Body Mass Index ; Case-Control Studies ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; ethnology ; Ethnic Groups ; Glucose Tolerance Test ; Humans ; Insulin ; Insulin Resistance ; Insulin-Secreting Cells ; Lipids ; Obesity ; Overweight ; Risk Factors ; Socioeconomic Factors

Objective To analyze the mutations of F12 gene in one pedigree with congenital factor FⅫ(FⅫ)deficiency, and investigate the molecular mechanisms of FⅫ deficiency.Methods Pedigree investigation.In February 2015,a patient with hereditary FⅫdeficiency was admitted to the Third Clinical College of Wenzhou Medical University.Activated partial thromboplastin time(APTT), prothrombin time (PT),FⅫactivity(FⅫ:C),FⅫ antigen(FⅫ:Ag)and other coagulant parameters were tested in the proband and his family members.5′and 3′UTR, all exons and their exon-intron boundaries of F12 gene were analyzed by direct sequencing.The detected mutations were confirmed by reverse sequencing.The conserved amino acids were analyzed by ClustalX-2.1-win software, and four bioinformatics softwares (PolyPhen-2,PROVEAN,SIFT and MutationTaster)were also used to analyze the effect of mutations on protein function.Results The proband and her younger brother showed a markedly prolonged APTT which were 116.4 s and 101.3 s, while her father had slightly prolonged APTT, and other family members were normal.The FⅫ:C and FⅫ:Ag of family members were also decreased(the proband,2.0% and 1.0%;her younger brother,2.0% and 1.0%; her father,18.0% and 13.0%).The phenotype of all members was consistent with cross-reactive material(CRM)negative.Nucleotide sequencing analysis showed that the proband and her younger brother had missense mutations in the F 12 gene, including one homozygous mutation c.1681G>A(p.Gly542Ser)and a commonly reported single nucleotide polymorphism site within the promoter region of the F12 gene(46T/T).Sequencing results from the proband's parents and son demonstrated them as carriers of a heterozygous missense mutation.The proband's husband was normal and with 46C/C in the promoter region.The ClustalX-2.1-win results indicated that the Gly542 was highly conserved among the homologousspecies.The predicting outcomes of the four bioinformatics softwares were the same,the PolyPhen-2(score 1.000)and PROVEAN(score -4.975)both declared p.Gly542Ser was a harmful mutation.The SIFT(score 0.00)and the MutationTaster(score 0.999)manifested the mutation could affect the protein funtion.Conclusions c.1681G>A(p.Gly542Ser)in exon 14 and 46T/T were related with the significant decrease of the FⅫlevel of this pedigree of hereditary FⅫ deficiency.

Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.

Objective:To evaluate the clinical value of sonography based volume computer aided display heart (SonoVCADheart) in the display of key diagnostic elements in basic fetal echocardiographic views.Methods:4D volume data based on fetal four-chamber view of 80 singleton fetuses (including 57 normal fetuses and 23 fetuses with heart abnormalities) were collected by using a volumetric probe from the Sir Run Run Shaw Hospital, Zhejiang University College of Medicine from January 8-22, 2019. Four to five volume datasets based on four-chamber view of the heart were rapidly acquired from each fetus. Three doctors (Doctor A: Engaged in fetal echocardiography diagnosis for more than 15 years, Doctor B and C: 6 months of regular fetal echocardiography training but lack of experience) with different fetal echocardiographic experiences performed off-line processing using SonoVCADheart combined tomographic ultrasound imaging (TUI) at different times to obtain eight standard echocardiographic diagnostic views, and to score the elementary contents of each echocardiographic view. The scores of the same doctor at different times, the scores of less experienced doctors and experienced doctor, and the scores between normal and abnormal fetuses, and the time required for analysis and diagnosis among all doctors were analyzed and compared, respectively. The diagnostic coincidence rates of SonoVCADheart for fetuses with cardiac malformations were also assessed.Results:A total of 279 volume datasets obtained from 57 normal fetuses, an average of 4.89/fetus, and 109 volume datasets obtained from 23 fetuses with cardiovascular abnormalities, an average of 4.74/fetus, and all volume datasets were used for SonoVCADheart analysis. The volume percentage of all the elements in the 8 diagnostic views with image quality≥2 points shown by SonoVCADheart in the normal fetuses was about 70.61%-74.91%, in the abnormal fetuses was about 53.21%-55.96%. There were no significant differences in the scores between the same doctor at different times, the scores between inexperienced doctors, and the scores among experienced and less experienced doctors(all P>0.05). There were significant differences in the scores between normal and abnormal fetuses except for all of the superior and inferior vena cava view and the aorticarch view of doctors A and C(all P<0.05). The time required for experienced doctor A and inexperienced doctors B and C to obtain 8 diagnostic views and to complete the diagnosis was statistically significant ( P<0.05). There was no significant difference in the time required for the inexperienced doctors B and C to obtain 8 diagnostic sections and to complete the diagnosis ( P>0.05). There was statistically significant difference in the diagnosis time required for doctor A using SonoVCADheart and two-dimensional echocardiography in fetuses with cardiovascular malformations ( P<0.05). However, there was no statistically significant difference in the time required in the normal fetus between SonoVCADheart and two-dimensional echocardiography ( P>0.05). The diagnostic coincidence rate of SonoVCADheart for fetuses with cardiac malformations is about 89.91%-90.83%. Conclusions:SonoVCADheart is a repeatable and stable novel fetal heart processing tool enabling displaying eight standard diagnostic sections of the fetal heart, and has potential clinical application value in the standardization of image acquisition and sequence display.

Objective: To explore the feasibility of Sonography based Volume Computer Aided Display Heart (SonoVCADheart) in the display of the fetal ventricular outflow views, and compare diameters of fetal aorta (AO) and pulmonary artery (PA) measured by two-dimensional echocardiography (2DE) and SonoVCADheart. Methods: Eighty singleton fetuses in the second and third trimesters of pregnancy in January 2019 in Sir Run Run Shaw Hospital, Zhejiang University College of Medicine were enrolled. Conventional 2DE examinations were performed. The volume datasets were analyzed offline using the new automatic image processing software SonoVCADheart. The diameters of AO and PA were measured by 2DE and SonoVCADheart, respectively. Pearson correlation analysis was used to evaluate the correlation between the two methods. The consistency of the two methods was verified by Bland-Altman analysis, and he reliability of SonoVCADheart was assessed using the intraclass correlation coefficient (ICC). Results: Fetal ventricular outflow views were successfully obtained using SonoVCADheart in 73(91.2%) of 80 fetuses. There were good correlations between the two methods for measuring the diameters of AO and PA (r=0.953, 0.971; all P<0.001). The 95% agreement limits of AO and PA were (-0.669, 0.568)mm and (-0.632, 0.580)mm, respectively. ICC demonstrated that SonoVCADheart achieved great repeatability both between and within observers. Conclusions SonoVCADheart may have potentials for the quantitative evaluation of fetal ventricular outflow tracts with its good repeatability and reliability.