Objective:To discuss the effect of downregulating microRNA-208a(miR-208a)on the resistance of the colorectal cancer cells to 5-fluorouracil(5-FU),and to clarify its related molecular mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-208a and secreted frizzled-related protein 1(SFRP1)mRNA in the 5-FU-resistant colorectal cancer cell line HT-29/5-FU and its parent HT-29 cells.The HT-29/5-FU cells were transfected with miR-208a inhibitor plasmid and its negative control plasmid(inhibitor-NC),and SFRP1 small interfering RNA(si-SFRP1)and its negative control plasmid(si-NC),either separately or in combination,followed by treatment with 5-FU.The cells were divided into inhibitor-NC group,miR-208a inhibitor group,miR-208a inhibitor+si-NC group,and miR-208a inhibitor+si-SFRP1 group.MTT assay was used to detect the proliferation activities of the cells and the resistance indexes were calculated;Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptotic rates of the cells after treated with different concentrations of 5-FU;Western blotting method was used to detect the expression levels of SFRP1,β-catenin,P-glycoprotein(P-gp),and ATP-binding cassette subfamily B member 1(ABCB1)proteins in the cells in various groups;dual-luciferase reporter gene assay was used to validate the targeting relationship between miR-208a and SFRP1.Results:Compared with HT-29 cells,the expression level of miR-208a in the HT-29/5-FU cells was increased(P＜0.05),and the expression level of SFRP1 mRNA was decreased(P＜0.05).Compared with inhibitor-NC group,the proliferation activity of the cells in miR-208a inhibitor group was decreased(P＜0.05),the resistance index was decreased,the apoptotic rate was increased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were decreased(P＜0.05).The dual-luciferase reporter gene assay results showed that SFRP1 was a target gene of miR-208a and miR-208a could negatively regulate the expression of SFRP1.Compared with miR-208a inhibitor+si-NC group,the proliferation activity of the cells in miR-208a inhibitor+si-SFRP1 group was increased(P＜0.05),the resistance index was increased,the apoptotic rate was decreased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were increased(P＜0.05).Conclusion:Downregulation of miR-208a can improve the resistance of the HT-29/5-FU cells to 5-FU by targeting and upregulating the SFRP1 expression,thereby inhibiting the transmission of the Wnt signaling pathway.

Objective To investigate the application value of low-molecular-weight heparin calcium combined with alprostadil in the treatment of hypoparathyroidism(HP)after thyroid surgery.Methods Sixty patients with HP after thyroid surgery were randomly divided into two groups:study group(30 cases)and control group(30 cases).The study group received low-molecular-weight heparin calcium and alprostadil,along with oral calcium supplements and calcitriol or an intravenous infusion of calcium gluconate,for three consecutive days after surgery.The control group received only oral calcium supplements and calcitriol or intravenous calcium gluconate.The values and changes in parathyroid hormone(PTH),postoperative drainage volume,and postoperative complications were compared between two groups.Results There were no significant difference in PTH levels at 3 days,1 week,1 month,and 6 months after surgery between two groups(P＞0.05).The difference in PTH levels between 3 days and 1 day after surgery was(6.77±2.61)ng/L in study group and(1.29±1.12)ng/L in control group,with a significant difference(P＜0.05).The difference in PTH levels between 1 week and 1 day after surgery was(10.51±3.66)ng/L in study group and(2.65±2.17)ng/L in control group(P＜0.05).There were no significant difference in PTH levels at 1 month,6 months,and 1 day after surgery(P＞0.05).Comparison of postoperative drainage volume:The postoperative drainage volume of study group was(90.07±22.24)ml,and that of control group was(88.20±24.09)ml,with no statistical significance(P＞0.05).There was no significant difference in postoperative complications between two groups(P＞0.05).Conclusion In cases of HP after total thyroidectomy,timely use of low-molecular-weight heparin calcium and alprostadil can accelerate the recovery of PTH and reduce patient numbness,which is safe,effective,and worthy of clinical promotion.

Bilateral vestibulopathy（BVP） is one of the common diseases in the vestibular nervous system, with an incidence rate of about 4%-7% in the population, which can lead to a variety of body dysfunctions. At present, there are two main treatment methods for BVP. One is vestibular rehabilitation. However, only part of BVP patients can finally benefit from vestibular rehabilitation, and most patients will remain with permanent vestibular dysfunction. Benefiting from the maturity of cochlear implant technology, European and American countries took the lead in the development of vestibular prosthesis（VP） technology to restore the vestibular function in patients with BVP. This review will focus on the development history, principles, future applications and the related research progress of VP in China.
Humans ; Bilateral Vestibulopathy/therapy* ; Cochlear Implants ; Vestibule, Labyrinth ; Cochlear Implantation ; China

Objective: To test the mechanism and upstream pathway of outer hair cell apoptosis in Cadherin 23 (Cdh23) gene mutant mice. Method: The mutant Cdh23^erl/erl(erl) mice were collected as the study group, while the C57BL/6J (B6) mice were chosen as the control group. A total of 70 mice per group were used in this study. The study group and control group underwent auditory-evoked brainstem response (ABR) tests at the same age. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect outer hair cell(OHC) apoptosis. The qRT-PCR was conducted to test the expression of ER stress markers immunoglobulin-binding protein (BiP) and C/EBP homologous protein (CHOP) mRNA. The expression and location of BiP and CHOP protein in OHC were detected by immunostaining. The expression of BiP protein in cochleae was identified by Western blot. The expression and location of CDH23 protein in OHC were discovered by immunostaining. Results: The ABR thresholds in erl mice were significantly higher than those in B6 mice at the age of 1 and 3 months (both P<0.05). The surface preparation with TUNEL staining confirmed OHC apoptosis in erl mouse cochleae which showed a higher TUNEL positive cell ratio than B6 mouse(t=11.291, P<0.01). The ER stress marker Bip and Chop mRNA were upregulated in the erl mouse inner ear, when compared with those in the B6 mouse(both P<0.05). The BiP protein extracted from the erl mouse cochleae was significantly higher than that of B6 mouse measured by Western blot (t=3.66, P=0.02). Immunostaining showed that BiP and CHOP were highly detected in the OHC in erl mouse cochleae, and was mainly detected in the perinuclear region of OHC. However, a bare BiP and CHOP signal were shown in B6 mouse cochleae. The CDH23 protein was specifically localized at the top of the OHC in B6 mice, indicating the localization of the tip links in hair bundle stereocilia. On the contrary, the CDH23^erl protein was found to be localized from the top to the nuclei of the OHC in erl mice. Portions of the CDH23^erl proteins failed to reach the top of the hair bundles and remained in the OHC cytoplasm. Conclusion As the downstream response of the Cdh23 gene mutation, portions of the mutant CDH23^erl protein was accumulated in ER lumen resulting in the increase of ER loading and ultimately triggered ER stress and hair cell apoptosis in erl mouse cochleae.

Objective: To observe the parameters of the results of suppression head impulse paradigm (SHIMP) in healthy adults, and to provide reference for evaluating vestibular oculomotor reflex function in patients with peripheral vertigo. Methods: Fifty healthy adults, 22 males and 28 females, aged from 23-65 years, with an average age of (38.5±11.6) years, were recruited from January to March 2018. Parameters provided by the video head pulse software included the gains, the latency and the peak velocity of saccades, and comparison was made with head impulse paradigm (HIMP). Results: All subjects were elicited anti-compensatory saccades in SHIMP. The normal values of left and right gains were 1.02 and 1.10 in HIMP, and 0.93 and 1.01 in SHIMP respectively. The left and right saccades latency were (201.1± 50.8)ms and (187.0± 42.9)ms, and the peak saccadic velocity were (302.7±58.5)°/s and (291.5±46.5)°/s in SHIMP; there were small but significant difference between two sides about gains in HIMP and SHIMP, as well as latency in SHIMP(P<0.05); there were small but significant difference between HIMP and SHIMP about gains in ipsilateral(P<0.01); there were no significant difference between two sides about peak saccadic velocity in SHIMP(P>0.05). Conclusions SHIMP can be used for the examination of vestibular oculomotor reflex function in adult population. It is easy to be operated and is convenient for clinical application. Combined with head pulse test, the function of the semicircular canal can be evaluated together.

Objective To explore the feasibility of hybrid procedure of percutaneous puncture for residual shunt after ventricular septal defect(VSD) repair.Methods From March 2015,we did percutaneous punctured transcatheter device closure of residual shunt after VSD repair for four patients.The patients included 2 males and 2 females with age from 4 years 9 months to 11 years 7 months and weight from 18 kg to44 kg.Four patients are diagnosed for4 years after VSD repair,3 years after tetralogy of Fallot (TOF),3 years after coarctation of aorta(CoA) combined with VSD,and 1 year after VSD repair,respectively.The withsize of residual shunt from 4.8 mm to 6.8 mm.Residual shunt are perimembranous,conoventricular and intracristal.Punctured directly through the right ventricular surface into the right ventricle in the 4-th or 5th intercostal space of the left sternal border.Next,extracted the guide wire after inserting a conveyor tube.Subsequently,7-10 mm VSD closure device were delivered via the conveyor tube.Results All the four operations were succeeded.A total of four closure devices were placed.Two of them are equilateral devices,and two are decentered devices,with size from 7mm to 9mm.Echocardiographyexamination revealed no significant pericardial effusion.Post-operative ultrasonic cardiogram showed the VSD closure device on site,with no pericardia1 effusion and no obvious morphological abnormalities of the valve.For one-year follow-up,the cardiac functionsof all patients were well.Conclusion Percutaneous punctured transcatheter device closure is a new,efficient and reliable method of treatment for children withresidual shunt after VSD repair.This method also can extend and offer a new avenue for valvular prosthesis and Perimembranous VSD.

Objective To explore the feasibility of percutaneous puncture for ventricular septal defect(VSD).Methods From March 2015,we did percutaneous punctured transcatheter device closure of VSD for 22 patients.The patients included 15 males and 7 females with age from 2 years 6 months to 11 years 7 months, weight from 13.5 kg to 44.0 kg.Among 22 pa-tients,6 were diagnosed with residual shunt after VSD repair, with size of residual shunt from 4.8 mm to 7.0 mm.Residual shunts are perimembranous, conoventricular and intracristal.The other 16 patients were diagnosed with isolated VSD,with de-fect size from 3.5 to 5.1 mm.For all patients,we punctured directly through the right ventricular surface into the right ventri-cle in the 3th or 4th intercostal space of the left sternal border.Next, we succeeded to extract the guide wire after inserting a conveyor tube for 21 patients.Subsequently,7-10 mm VSD closure device were delivered via the conveyor tube.Only one patient was changed to perform occlusion of VSD through chest small incision due to improper selection of punctured position. Results A total of 21 closure devices were placed for 21 patients.18 of them are equilateral device,and 3 are decentered de-vice,with size from 5 mm to 10 mm.There were small amount of pericardial effusion in 4 patients using TEE examination,and no future treat was performed after observation.For one-year follow-up, all patients have recovered very well.Conclusion Percutaneous punctured transcatheter device closure fits for treatment for children with residual shunt after VSD repair and isola-ted VSD.It has a good recent result.

Objective: To observe the features of air-conducted sound elicited ocular vestibular-evoked myogenic potential(ACS-oVEMP) and cervical vestibular-evoked myogenic potential(ACS-cVEMP) in patients with Meniere disease (MD). To analyze the relationship between air-conducted sound elicited vestibular-evoked myogenic potentials (ACS-VEMP) responses and clinical stages of disease, as well as its clinical application of cervical and ocular vestibular evoked myogenic potentials in MD. Method: Fifty six patients with MD and 50 normal subjects (100 ears) were recruited for conventional cVEMP and oVEMP examinations. Grades of vestibular function were also collected for patients with MD. The relationship between VEMPs abnormity, grades of vestibular function and clinical stages of MD were analyzed. Results: The abnormal rates of cVEMP and oVEMP in MD patients were 57.1% (32/56) and 64.3% (36/56), which were significantly higher than those in normal subjects respectively (χ²=22.286, P=0.000; χ²=15.217, P=0.000). The abnormal rates of cVEMP and oVEMP in MD patients of stage Ⅰ to stage Ⅳ were 20.0% (1/5) and 40.0% (2/5), 50.0% (9/18) and 50.0% (9/18), 59.3% (16/27) and 70.4% (19/27), and 100.0% (6/6) and 100.0% (6/6) respectively. There was a significant difference in cVEMP abnormity between four stages of MD patients (P=0.046). Significant correlation was found between clinical stages and the grades of vestibular dysfunction (r_s=0.417, P=0.001). Conclusions Dysfunction of vestibular otolithic organs and their input pathways in patients with MD can be detected by cVEMP and oVEMP tests. The abnormal rates of VEMP could show an gradually increasing trend with the development of MD stages. And the extent of vestibular lesions could be detected by cVEMP and oVEMP tests, which may provide a reference for clinical staging of MD.

Objective To develop a new method to detect A (TA)n TAA polymorphism in the UGT1A1 gene promoter by fluo‐rescence real‐time quantitative PCR (RQ‐PCR) .Methods Genomic DNA was extracted from peripheral blood in 16 patients with Gilbert′s syndrome and 66 healthy individuals .The polymorphic A(TA)n TAA sequence in the UGT1A1 gene promoter was deter‐mined by DNA sequencing .A pair of primers and two TaqMan probes labeled with either 5′FAM or VIC reporter dye incorporated a 3′minor groove binder were designed .The A(TA)n TAA polymorphisms in the UGT1A1 gene promoter were identified by RQ‐PCR for all research subjects .The sensitivity and specificity of RQ‐PCR for detecting the A(TA)nTAA polymorphisms were veri‐fied by DNA sequencing method .Results The homozygous A(TA)7TAA polymorphism was found in the promoter region of the UGT1A1 gene in all 16 patients with Gilbert′s syndrome by using RQ‐PCR .The homozygous A(TA)6TAA polymorphism was foundin46healthysubjects,whiletheheterozygousA(TA)6TAA/A(TA)7TAApolymorphismwasfoundinother20healthysub‐jects .All A(TA)nTAA polymorphisms in the promoter region of the UGT1A1 gene identified by RQ‐PCR were consistent with that of DNA sequencing .Conclusion It is a sensitive ,specific and simple method to detect the A (TA)n TAA polymorphisms in the promoter region of the UGT1A1 gene by RQ‐PCR ,which can be promoted and applied in clinic .

OBJECTIVETo compare the efficacy of three methods for forensic diatom test, namely strong acid digestion-centrifuge enrichment-light microscopy (SD-CE-LM), microwave digestion-membrane filtration-automated scanning electron microscopy (MD-ME-SEM), and microwave digestion-membrane filtration-light microscopy (MD-MF-LM).

METHODSSixty samples were randomly divided into 3 groups for diatom test using three methods, and the sample preparation time, degree of digestion and recovery rate of diatoms were compared.

RESULTSThe sample preparation time was the shortest with MD-MF-LM and the longest with SD-CE-LM (P<0.05). MD-ME-SEM and MD-MF-LM allowed more thorough tissue digestion than SD-CE-LM. MD-ME-SEM resulted in the highest total recovery rate of diatom, followed by MD-MF-LM and then by SD-CE-LM (P<0.05); the recover rate of different diatom species was the highest with MD-ME-SEM, followed by MD-MF-LM and SD-CE-LM (P<0.05).

CONCLUSIONSD-CE-LM has a low recovery rate of diatoms especially for those with lengths shorter than 40 µm or densities less than 1/5. With a high recovery rate and accuracy in diatom test, MD-ME-SEM is suitable for diagnosis of suspected drowning cases. MD-MF-LM is highly efficient, sensitive and convenient for forensic diatom test.

Centrifugation ; Diatoms ; isolation & purification ; Drowning ; Forensic Sciences ; methods ; Humans ; Microscopy ; Microscopy, Electron, Scanning ; Microwaves ; Specimen Handling