Function of IL-33 in Wound Age Estimation of Skin Wounds in Mice.

ABSTRACT: Objective To explore the application value of interleukin-33 （IL-33） in wound age estimation in forensic practice by observing the sequential changes of IL-33 after skin wound. Methods Skin wound models were generated on the back of mice with a round file of 5 mm in diameter. Skin samples of the same size were taken from the same parts of mice in control group and injury group 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d and 10 d after skin wound. Hematoxylin-eosin （HE） staining method was applied to observe the morphological changes in the recovering process after skin wound. Western blotting, immunohistochemistry staining and double immunofluorescence staining methods were applied to detect the expression changes of IL-33 in the skin wound samples. Results The results of Western blotting showed that the expression of IL-33 protein decreased slightly at 3 h after skin wound, increased gradually at 6 h after skin wound, and reached the peak value at 3 d, then decreased gradually. Immunohistochemistry staining results showed that faint positive expression of IL-33 was observed in epidermis, hair follicles, sebaceous glands and dermal resident cells of the control group skin. The positive cell rate of IL-33 increased at 3 h after skin wound and reached the peak value at 3 d, then decreased gradually. The results of double immunofluorescence staining showed that the majority of IL-33 positive cells from 1 d to 3 d after wound were macrophages, while the majority of IL-33 positive cells from 5 d to 7 d after wound were myofibroblasts. In addition, the results of HE staining showed that the wound healing process of the skin wound model was consistent with the pathological development law of inflammation. Conclusion IL-33 could become a reference index for wound age estimation of skin wound in forensic practice.

Application of Virtual Anatomy Technology in Postmortem Examination of Medical Dispute Cases.

ABSTRACT: Objective To discuss the application value of CT scanning technology in cause of death determination of medical dispute cases. Methods From July 2017 to December 2018, postmortem CT imaging data of 12 medical dispute cases were collected. CT imaging diagnosis results and anatomy findings as well as differences between antemortem and postmortem CT diagnosis were compared. The advantages and disadvantages of CT routine tests of the cadavers in terms of the diagnosis of disease and damage were analyzed. Results The comparison between CT imaging diagnosis and anatomical findings showed that CT scans had advantages in the diagnosis of disease and damage with large differences in density changes, such as atelectasis, pneumonia, calcification, fracture and hemorrhage, etc. The comparison of CT diagnosis in antemortem and postmortem examination showed that the cadavers of medical dispute cases were well preserved and that postmortem CT scan was meaningful for the diagnosis of antemortem diseases. Conclusion Virtual anatomy technology has a relatively high application value in postmortem examination of medical dispute cases. It can provide effective information for the appraisers before the autopsy and can also provide a reference for cause of death analysis when the anatomy cannot be performed.

Expression of Annexin A1 during Skin Incised Wound Healing in Mice.

OBJECTIVES: To investigate the expression changes of annexin A1 （ANXA1） during the process of skin incision healing, and to explore its expression and function during skin injury repair. METHODS: The skin injury model of mice was prepared, and skin tissues of the controls and the injured group at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d after injuries were taken. The morphological changes of the wound were observed by hematoxylin-eosin （HE） staining, and the expression of ANXA1 was detected by immunohistochemistry （IHC） and Western blotting. RESULTS: HE staining showed normal healing of skin wounds. IHC results revealed that ANXA1 was expressed in the epidermis, hair follicle, sebaceous gland and vascular endothelium. In the injured group, the expression of ANXA1 was enhanced in epidermis and skin appendages around the wound 6-12 h after injury, and ANXA1 was also highly expressed in neutrophils and a small number of mononuclear cells. ANXA1 was mainly positively expressed in monocytes, neovascular endothelial cells and fibroblasts, and small amount of fibroblasts at 1-3 d, 5-10 d, and 14 d after injury, respectively. Western blotting showed that, compared with the controls, the expression of ANXA1 was significantly increased at 6 h after injury, peaked at 1 d, and then decreased gradually in the injured group. CONCLUSIONS: ANXA1 may be involved in the regulation of skin damage repair, with time-dependent expression during skin wound healing, and thus is expected to be a biological marker for inferring the wound formation time.

Time-dependent Expression and Distribution of AChE during the Skin Incised Wound Healing in Mice.

ABSTRACT: Objective To study the time-dependent expression and distribution of acetylcholinesterase （AChE） during skin incised wound healing in mice, and discuss its effect in wound healing as well as the feasibility of using it as a reference index for wound age estimation. Methods A total of 45 C57BL/KsJ mice were randomly divided into one control group and eight incised groups. The skin incised wound model was established in the incised groups with samples of skin wounds taken at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d post-injury respectively, while the uninjured skin tissue was extracted in the control group. Expression and distribution of AChE in skin samples were detected by immunohistochemistry, double immunofluorescence and Western blotting. Results Immunohistochemistry results indicated that AChE was mainly detected in infiltrating polymorphonuclear cells （PMNs） 6 to 12 h post-injury. A large number of AChE-positive mononuclear cells （MNCs） were observed 1 to 3 d post-injury. The AChE-positive cells were mainly fibroblastic cells （FBCs） 5 to 14 d post-injury. The ratio of the AChE-positive cells increased initially 6 h post-injury, and reached the peak at 1 d post-injury. Double immunofluorescent staining showed that the majority of AChE-positive MNCs and FBCs expressed macrophage marker and myofibroblast marker, respectively. Western blotting results showed that the relative expression level of AChE in the incised group was higher than that in the control group averagely, reached the peak at 1 d post-injury, then reached a second peak at 7 d post-injury. Conclusion The expression of AChE is found in PMNs, macrophages and myofibroblast during skin wound healing, which indicates it might be involved in the adjustment of inflammatory response and fibrotic repair after injury. Moreover, combined use of various methods for the detection of the expression of AChE would provide reference for skin wound age estimation.

Relationship between the Number of Neutrophils and Myofibroblasts during Diabetic Wound Healing and Wound Age.

ABSTRACT: Objective To investigate the sequential changes of the number of neutrophils and myofibroblasts during diabetic wound healing, and discuss its application value in wound age estimation. Methods Diabetic DB mice and mice of the same age in the normal control group were selected, a wound healing model was established, wound samples were taken at different time points, while the number of neutrophils and myofibroblasts during diabetic wound healing were determined by immunohistochemical staining technique. Results The number of infiltrated neutrophils in the wounds of control and diabetic groups reached the peak respectively at 12 h and 5 d after injury. Compared with the control group, the number of neutrophils in the diabetic group decreased significantly from 6 h to 1 d after injury, but increased markedly from 5 d to 14 d. From 5 d to 10 d after injury, the average number of neutrophils at high magnification in wounds of the diabetic group was over 30, while that of neutrophils in wounds of the control group was less than 20. Myofibroblasts appeared in wounds from 3 d to 14 d after injury in the control group and from 5 d to 14 d after injury in the diabetic group. The difference in the number of myofibroblasts in wounds between control group and diabetic group from 3 to 7 d after injury had statistical significance. Conclusion In comparison with normal wound healing, the number of neutrophils and myofibroblasts during diabetic wound healing shows different sequential changes. The results of this study can provide reference for wound age estimation of patients with severe diabetes.

[Relationship between FoxO1 Expression and Wound Age during Skin Incised Wound Healing].

OBJECTIVES: To investigate FoxO1 expression and its time-dependent changes during the skin incised wound healing. METHODS: After the establishment of the skin incised wound model in mice, the FoxO1 expression of skin in different time periods was detected by immunohistochemistry and Western blotting. RESULTS: Immunohistochemistry staining showed that FoxO1 was weakly expressed in a few fibroblasts of epidermis, hair follicles, sebaceous glands, vessel endothelium and dermis in the control group. The FoxO1 expression was enhanced in the epidermis and skin appendages around the wound during 6-12 h after injury, which could be detected in the infiltrating neutrophils and a small number of monocytes. FoxO1 was mainly expressed in monocytes during 1-3 d after injury, and in neovascular endothelial cells and fibroblasts during 5-10 d. On the 14th day after injury, the FoxO1 expression still could be detected in a few fibroblasts. The Western blotting results showed that the FoxO1 expression quantity of the tissue samples in injury group was higher than in control group. The FoxO1 expression peaked at 12 h and 7 d after injury. CONCLUSIONS: FoxO1 is time-dependently expressed in skin wound healing, which can be a useful marker for wound age determination.

Construction of HEK293 cells stably expressing wild-type organic anion transporting polypeptide 1B1 (OATP1B1*1a) and variant OATP1B1*1b and OATP1B1*15.

A transgenic cell line stably expressing the human organic anion transporting polypeptide (OATP1B1) was established. Human Embryonic Kidney 293 (HEK293) cell line stably expressing OATP1B1*1a sequence was amplified through PCR with the extracted total RNA as templates from human liver, then subcloned into the plasmid pMD19-T and verified by sequencing. OATP1B1*1b/OATP1B1*15 mutant sequences were obtained by site-directed mutation PCR with pMD19-T/ OATP1B1*1a as templates. The plasmids pcDNA3.1(+)/OATP1B1*1a, *1b and *15 were constructed and transfected into HEK293 cell line using Lipofectamine 2000 transfection reagent. Several stable transfected clones were obtained after selection with G418. Using rosuvastatin as a probe substrate of OATP1B1, the intracellular rosuvastatin accumulation in HEK293 and HEK-OATP1B1*1a, *1b and *15 monoclone cells were validated by a ultra-performance liquid chromatography-tandem mass spectrometry. OATP1B1 mRNA and protein expression were detected by RT-PCR and Western blot, respectively. The results from RT-PCR, rosuvastatin uptake and Western blot assay indicated that human OATP1B1 was highly expressed in transfected cells compared with controls. The HEK-293 cell lines stably expressing human OATP1B1-wild and variant (HEK-OATP1B1, *1b and *15) are potential models to study drug transport in vitro.

Identification of CYP2C19 inhibitors from phytochemicals using the recombinant human enzyme model.

The aim of the present study was to develop the recombinant insect cell-expressed protein as an in vitro model for inhibitors screening for human cytochrome P450 2C19 (CYP2C19), and to use the model to investigate the inhibition effect of three phytochemicals on CYP2C19 in vitro. Omeprazole was applied as the probe substrate. The estimated inhibitory constant (K(i)) of ticlopidine and fluvoxamine were 0.64 +/- 0.025 microM and 0.29 +/- 0.090 microM, respectively. After co-incubation with ticlopidine or fluvoxamine, the mean omeprazole Michaelis-Menten constant (K(m)) increased from 4.99 +/- 0.22 microM to 16.25 +/- 1.22 microM or 19.20 +/- 1.73 microM, respectively, while omeprazole's mean V(max) did not vary much. Both ticlopidine and fluvoxamine were competitive inhibitors of CYP2C19. The IC50 of three phytochemicals, isoalantolactone, curcumol and schisandrin A was determined as 38.91 microM, 121.0 microM and 86.41 microM, and the K(i) as 5.02 +/- 1.04 microM, 35.84 +/- 8.95 microM, and 4.46 +/- 0.017 microM, respectively. The in vitro model for inhibitor screening established using recombinant CYP2C19 could be used to assess the inhibition potential of drug candidates. Isoalantolactone and schisandrin A are potent inhibitors of CYP2C19, while curcumol is a moderate potent inhibitor of CYP2C19.

[Preliminary analysis of risk factors of tinnitus related to female menopause].

Objective: To investigate the possible pathogenesis and possible risk factors of tinnitus related to female menopause. Methods: From April 2016 to October 2016, 59 female patients with menopausal syndrome were diagnosed in the menopause comprehensive management clinic. Tinnitus and menopause questionnaires were conducted, based on whether having tinnitus, those patients were divided into two groups: tinnitus group and no tinnitus group. Age, body mass index (BMI), Kupperman menopausal index (KMI) score, follicle-stimulating hormone (FSH) level of patients in the two groups were analyzed. Menopausal symptoms, related medical history and possible related factors of tinnitus were statistically analyzed. Results: A total of 59 cases were collected, 22 of which were accompanied by tinnitus. The incidence of idiopathic tinnitus was 35.1% (20/57) because 2 cases of thyroid related tinnitus with clear etiology were removed. Complete data were obtained from 17 of 20 patients with idiopathic tinnitus and 26 of 37 patients without tinnitus. Age, BMI, menopause KMI score, hormone level, menopause symptoms and possible factors related to tinnitus were statistically analyzed between the two groups, and the incidence of headache was statistically different between the two groups (χ2=9.098, P=0.003), but no other factors were statistically significant(P>0.05). The severity of insomnia and tinnitus were further analyzed (χ2=2.841, P=0.417), and there was no significant difference between the two groups. Conclusion: Headache history may be one of the high risk factors for the occurrence of menopausal tinnitus.

Construction and application of double-transfected cells expressing the human transporter P-glycoprotein and cytochrome P450 3A4.

Intestinal P-glycoprotein (P-gp) and cytochrome P450 (CYP) enzymes are known to influence oral bioavailabilities of drugs. Recombinant plasmids pcDNA3.1/Hypgro/CYP3A4 were transfected into MDCK and MDCK-MDR1 cells to construct the single-transfected cell line MDCK-CYP3A4 and double-transfected cell line MDCK-MDR1/CYP3A4. The expression of CYP3A4 in the double-transfected cell line was determined by Western blot and its activity was detected by the metabolism assays of three substrates of CYP3A4, which were 7-benzyloxy-4-trifluoro-methylcoumarin (BFC), testosterone and midazolam. In addition, the selection of monoclones with high CYP3A4 activities in the single-tranfected cell line was performed by the P450 Glo CYP3A4 assay. Through MTT assay, the interaction between P-gp and CYP3A4 was preliminarily determined based on the changes of IC50 values. The results showed that paclitaxel detoxified in the single-transfected MDCK-MDR1 cell because of P-gp efflux. And it was also less toxic in the single-transfected CYP3A4 cell line due to the metabolism by CYP3A4. In the double-transfected MDCK-MDR1/CYP3A4 cell line, the toxicity decreased dramatically because of the interplay between P-gp and CYP3A4. Therefore, the cell model could be applied to study the toxicity and detoxification of chemicals due to the metabolism by CYP3A4 and the efflux through P-gp.

[Reclassification of flat type sudden deafness].

Objective: To reclassify the flat type sudden deafness according to the types of audiogram shape, and to explore the correlation between different pattern of hearing loss and prognosis. Methods: All of 1 024 patients with unilateral sudden deafness (492 males and 532 females, aged from 19 to 65 years, with an average age of 41.2 years old) admitted to 33 hospitals nationwide from August 2007 to October 2011 were divided into four types according to Chinese Guideline of Sudden Deafness(2015): low-frequency, high-frequency, flat and total deafness. Then, 402 patients with flat type sudden deafness were further divided into ascending type, descending type and consistent type according to the audiogram shapes. First, we compared the clinical characteristics and prognosis among these three subtypes of flat deafness, then compared the clinical characteristics and prognosis between ascending flat deafness and low-frequency deafness, descending flat deafness and high-frequency deafness, consistent flat deafness and total deafness, explored the factors related to the prognosis of flat deafness. SPSS 21.0 software, ANOVA, χ2 test, t-test and Logistic regression were used to analyze the data. Results: The cure rates of flat ascending, flat descending and flat consistent sudden deafness groups were 70.7%, 17.1% and 34.0% respectively, with a statistically significant difference (χ2=33.984, P<0.001); However, there was no significant difference in age, sex and affected side (all P>0.05). The independent related factors for the recovery of flat type sudden deafness were as follows: whether there was dizziness [OR=0.459; 95% confidence interval (CI): 0.271-0.777], the type of audiogram shape (OR=0.721; 95%CI: 0.530-0.981), and days from onset to therapy (OR=0.903, 95%CI: 0.835-0.978), all of which had P values<0.05. There was no significant difference in the cure rates between ascending flat sudden deafness and low-frequency descending sudden deafness, descending flat sudden deafness and high-frequency descending sudden deafness (all P>0.05). The pure tone average(PTA) of flat consistent sudden deafness and total deafness were (69.1±18.9) and (101.7±17.7) dB HL, respectively, with a statistically significant difference (t=20.890, P<0.001), and the cure rates were 34.0% and 14.5%, respectively, with a statistically significant difference (χ2=29.012, P<0.001). Conclusion: According to the audiogram shape, the flat type sudden deafness can be further divided into ascending flat sudden deafness, descending flat sudden deafness and consistent flat sudden deafness, which can more effectively evaluate the prognosis. The cure rate of ascending flat sudden deafness is similar to that of low-frequency sudden deafness, and the prognosis is well; The cure rate of descending flat sudden deafness is similar to that of high-frequency descending sudden deafness, and the prognosis is poor. The cure rate of consistent flat sudden deafness is higher than that of total deafness. PTA plays an important role in the prognosis of consistent flat sudden deafness and total deafness. Total deafness can be regarded as a single type of sudden deafness.

Comparison of catalytical activity and stereoselectivity between the recombinant human cytochrome P450 2D6.1 and 2D6.10.

Polymorphisms of the cytochrome P450 2D6 (CYP2D6) gene play a major role in pharmacokinetic variability in human, while CYP2D6*10 is an important subtype in Asian people. In this study, the co-expression enzyme of human recombinant CYPOR, CYPb5 and CYP2D6.1 or CYP2D6.10 with the Bac-to-Bac system in baculovirus-infected insect cells was used to study the catalytical activity to imipramine metabolism and stereoselective metabolism of propranolol. The metabolites of imipramine were identified of hydroxyl imipramine and desipramine by LC-MS/MS. There are some differences between CYP2D6.1 and CYP2D6.10 activity. The kinetics parameters K(m), V(max), and CL(int) are 11.77 +/- 0.91 micromol/L, 0.4235 +/- 0.05 nmol/nmol CYP2D6.1/min and 3.60 x 10(-5) ml/min/nmol CYP2D6.1 (n = 3) for CYP2D6.1, respectively, and 9.05 +/- 0.87 micromol/L, 0.42 +/- 0.03 nmol/nmol CYP2D6.10/min, and 4.60 x 10(-5) ml/min/nmol CYP2D6.10 (n = 3) for CYP2D6.10. For propranolol, two metabolites were identified to be hydroxyl and N-desisopropylation propranolol by LC-MS/MS. When the substrate concentration was 0.20 micromol/L, CYP2D6.1 and CYP2D6.10 exhibited significant stereoseletivity. Furthermore, enantioselective formation has been detected. Both of CYP2D6.1 and CYP2D6.10 produced more hydroxyl propranolol from the R-(+)-isomer than from the S-(-)-isomer while there was no obvious difference for N-desisopropylation propranolol production between R-(+)- and S-(-)- isomer. In summary, there is a somewhat different catalytical activity and stereoselectivity between the human recombinant CYP2D6.1 and CYP2D6.10. The data we got will be helpful in preclinical research and clinical use of CYP2D6 substrates.

[The peripheral blood inflammatory markers in Ménière's disease patients with and without migraine].

Objective: To investigate the peripheral blood inflammatory markers including white blood cell count (WBC), monocytes, neutrophils, lymphocytes, platelets, neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), high-density lipoprotein(HDL-C), low-density lipoprotein and fibrinogen (FIB) in Ménière's disease (MD) patients with and without migraine, and to explore the relationship between the inflammatory response with MD and migraine. Methods: The general physical condition, clinical manifestations, pure-tone audiometry, and peripheral blood inflammatory markers of 92 unilateral MD patients who were hospitalized in Peking University People's Hospital for surgical treatment from January 2017 to January 2021 were continuously collected. Meanwhile, 50 healthy controls matched with age and sex were included, and their general physical conditions and peripheral blood inflammatory markers were also collected. This study consisted of two parts. First, the differences in epidemical characteristics and peripheral blood inflammatory markers between MD patients and healthy controls were compared by univariate analysis. Second, all 92 MD patients were divided into two subgroups according to whether they were accompanied by migraine. The clinical characteristics and peripheral blood inflammatory markers of MD patients with and without migraine were compared by univariate analysis. Thereafter, binary Logistic regression was used to analyze the related factors of whether MD patients were accompanied with migraine. Results: Compared with the healthy control group, the peripheral blood WBC, neutrophils and FIB of MD patients were significantly increased (all P<0.05). Compared with MD patients without migraine, MD patients with migraine had higher female prevalence, longer disease history, lower low-frequency hearing threshold, higher frequency of vertigo attacks and higher HDL-C levels (all P<0.05), meanwhile, female, frequency of vestibular attacks and HDL-C were independent related factors of whether MD patients were accompanied with migraine. Conclusion: The occurrence of MD and migraine may be related to the inflammatory response. The level of anti-inflammatory factors in the blood of MD patients with migraine are higher, suggesting that the inflammatory response status of MD patients with and without migraine is different.

Prevalence of anxiety in patients awaiting diagnostic procedures in an oncology center in Brazil.

OBJECTIVE: Anxiety disorders have been shown to undermine the quality of life of cancer patients. Unfortunately, medical professionals often neglect to screen for anxiety in their patients. The aim of the present study was to describe the prevalence of anxiety in patients awaiting diagnostic procedures in an oncology center waiting room, and to investigate possible relationships between anxiety and demographic and clinical variables. METHODS: A cross-sectional study was performed with 398 patients who completed a self-administered questionnaire containing the Hospital Anxiety and Depression Scale (HADS) and the State-Trait Anxiety Inventory (STAI). RESULTS: Results of the HADS indicated that 38% of participants had anxiety, while data from the STAI showed that 46% had either high state or trait anxiety. The most frequently cited source of anxiety was concern over test results. Age, gender, employment status, and education level were correlated with anxiety. CONCLUSIONS: The prevalence of anxiety is high among patients awaiting diagnostic procedures. Patients in the waiting room should be routinely screened for anxiety. Careful assessment and treatment of anxiety are important components in the care of patients with cancer.

[Gadolinium angiography of inner ear in patients with sudden deafness and its clinical analysis].

Objective: To observe the gadolinium imaging findings of inner ear in patients with sudden deafness and to analyze its clinical features. Methods: From November 2017 to July 2020, 21 patients with sudden deafness in the People's Hospital of Dongsheng District, Ordos City were selected as the research objects, including 14 males and 7 females, aged 36-76 years, with a median age of 50 years. The course of disease was 1-19 days, with an average of 5.5 days. The patients received audiology tests, laboratory examination, and intravenous gadolinium angiography, each of whom was scanned twice by 3D-FLAIR sequence: once before intravenous gadolinium injection, and once again 4.5-6.0 h after intravenous gadolinium injection. The following corresponding clinical treatment was given. The imaging manifestations and clinical features were observed. Results: Among 21 cases of sudden deafness in acute stage, the signal intensity of 11 cases was significantly higher than that of the contralateral ear, and 2 cases had vestibular labyrinthine hydrops. In laboratory examination, only 2 cases of total deafness had increased WBC count and faster erythrocyte sedimentation rate, and the rest had no abnormality. The hearing types of 21 patients with sudden deafness were: total deafness in 8 cases, flat decline in 10 cases, low frequency decline in 1 case, high frequency decline in 2 cases. The total effective rate was 57% (12/21). The hearing types of 11 patients with abnormal gadolinium angiography were total deafness in 5 cases, flat decline in 5 cases and high frequency decline in 1 case. The total effective rate was 64% (7/11). Conclusion: Gadolinium angiography is abnormal in some patients with sudden deafness, and the permeability of blood labyrinth barrier may be increased, which is worthy of further study.

Structural elucidation of metabolites of ginkgolic acid in rat liver microsomes by ultra-performance liquid chromatography/electrospray ionization tandem mass spectrometry and hydrogen/deuterium exchange.

Ginkgolic acids have been shown to possess allergenic as well as genotoxic and cytotoxic properties. The question arises whether the metabolism of ginkgolic acids in the liver could decrease or increase their toxicity. In this study, the in vitro metabolism of ginkgolic acid (15:1, GA), one component of ginkgo acids, was investigated as a model compound in Sprague-Dawley rat liver microsomes. The metabolites were analyzed by ultra-performance liquid chromatography coupled with photodiode array detector/negative-ion electrospray ionization tandem mass spectrometry (UPLC-PDA/ESI-MS/MS) and hydrogen/deuterium (H/D) exchange. The result showed that the benzene ring remained unchanged and the oxidations occurred at the side alkyl chain in rat liver microsomes. At least eight metabolites were found. Among them, six phase I metabolites were tentatively identified. This study might be useful for the investigation of toxicological mechanism of ginkgolic acids.

[The correlation between plasma fibrinogen level and different types of sudden sensorineural hearing loss].

Objective: To investigate the correlation between plasma fibrinogen level and the incidence of sudden hearing loss. Method: A total of 225 patients (age ranging from 18 to 70 years) with sudden sensorineural hearing loss(SSNHL) were selected. The disease onset was within 2 weeks. No patient received previous medical intervention. Depending on the types of auditory threshold curve, SSNHL was divided into low frequency descending type, high frequency descending type, flat desending type and total deafness type. Two hundred and twenty-nine cases with normal hearing and no inflammatory diseases in the same period was selected as control group. Fibrinogen level and Blood Routine Indexes were detected for the purpose of retrospective cohort study.Result: The levels of fibrinogen in SSNHL group(2.98±0.59) g/L were significantly higher than that in control group(2.66±0.36) g/L (P<0.01). According to the types of auditory threshold curve, SSNHL groups were divided into group A(55 cases, 24.44%), Group B(36 cases, 16.00%), Group C(43 cases,19.11%) and Group D(91 cases, 40.44%). The average fibrinogen levels before treatment were at(2.75±0.46)g/L, (3.16±0.61) g/L, (3.02±0.63) g/L and(3.03±0.63) g/L respectively. There was no significant difference in fibrinogen level(P=0.286) between group A and the control group. Fibrinogen levels of group B, group C and group D were significantly elevated compared to control group (P<0.01). Conclusion: The level of fibrinogen is not significantly correlated with development of sudden deafness of low frequency descending type. The incidence of high frequency descending type, flat descending type and profound deafness type are all correlated with the level of high fibrinogen at the onset of sudden deafness. The classification of sudden deafness based on audiogram curves could be of great significance for analysis of possible causes and selection of treatment options.

[Advances in the relevance of laryngopharyngeal reflux and tinnitus].

Tinnitus is the most common symptom in otorhinolaryngology. In the past, tinnitus was often localized in the auditory system lesions, causing a great amount of idiopathic tinnitus patients inadequately treated. At present, it is believed that the pathogenesis of tinnitus not only originates from auditory system itself, but also participates in systemic disorders. In recent years, investigating in the effects of laryngopharyngeal reflux on tinnitus is undergoing rapid progress. This review highlights the current state of relationship between laryngopharyngeal reflux and tinnitus, which may provide a new systemic insight into diagnosis and treatment on tinnitus as well as laryngopharyngeal reflux.

[Study on the effects of microRNA-203 on the invasion and apoptosis of laryngeal cancer cells via targeting LASP1].

Objective: To explore the role of microRNA-203 in laryngeal cancer and its underlying mechanism and clarify the relationship between microRNA-203 and LASP1.Method: microRNA-203 expression in laryngeal cancer tissues and paracancerous tissues was detected by quantitative real time-polymerase chain reaction（qRT-PCR）. The regulatory effects of microRNA-203 on invasion and apoptosis of laryngeal cancer cells were detected by Transwell assay and flow cytometry, respectively. Dual-luciferase reporter gene assay was performed to access the binding condition of microRNA-203 and LASP1. Both mRNA and protein levels of LASP1 in laryngeal cancer cells were detected after transfection with microRNA-203 mimic or microRNA-203 inhibitor by qRT-PCR and Western blot. Rescue experiments were finally performed to detect whether microRNA-203 regulates laryngeal cancer development via targeting LASP1. Result: microRNA-203 was lowly expressed in laryngeal cancer tissues and cell lines.Knockdown of microRNA-203 in Hep-2 cells can promote the invasiveness and inhibit apoptosis of laryngeal cancer cells. Subsequently,LASP1 was predicted to be the target gene of microRNA-203，which was further verified by dual-luciferase reporter gene assay.LASP1 expression was negatively regulated by microRNA-203. Furthermore，rescue experiments showed that microRNA-203 regulates invasion and apoptosis of laryngeal cancer cells via targeting LASP1. Conclusion： Low expression of microRNA-203 could promote the invasion and inhibit apoptosis of laryngeal cancer cells viainhibiting LASP1. microRNA-203 and LASP1 both play a very important role in the development of laryngeal cancer..

[Correlation analysis of incidence, season and temperature parameters of different types of sudden deafness].

Objective:The objective of this study was to investigate the correlation between the onset of different types of sudden sensorineural hearing loss（SSNHL） with temperature parameters and seasons. Method:We retrospectively reviewed the medical charts of 175 patients who were diagnosed as SSNHL, precisely collected the exact date and city of onset, confirmed the season, and obtained the meteorological data including maximum temperature（Tmax）, minimum temperature（Tmin）, mean temperature（T）, day-to-day change of mean temperature（ΔT）, and diurnal temperature range（Trange） at the same day, then analyzed the relation between season and temperature with the onset of different types of SSNHL. Result:There was a significant difference of Trange between different types of SSNHL（P=0.001）. Trange on the onset date of all-frequency SSNHL（including flat and profound type） was significantly higher than low and high frequency descending type（P=0.001, P<0.05 respectively）. Types of SSNHL had weak association with Trange groups（P=0.03, Cramer's V=0.220）. An increase of 1℃ in Trange increased the risk of flat type SSNHL by 23.9% and 16.5% compared with low and high frequency descending type, respectively, and for profound type, the risk was increased by 22.4% and 15.1%. No significant differences were observed between seasons and SSNHL types（P=0.666）. Conclusion:The incidence of different types of sudden sputum may be related to the worse temperature on the day, and has nothing to do with the disease season.