Objective:To explore the genetic and clinical characteristics of Guizhou patients with enlarged vestibular aqueduct（EVA） syndrome through combined SLC26A4 variant analysis and clinical phenotype analysis. Methods:Seventy-two EVA patients underwent comprehensive genetic testing using a multiplex PCR-based deafness gene panel and next-generation sequencing（NGS）. The audiological and temporal bone imaging characteristics were compared across mutation subtypes. Results:A total of 27 pathogenic loci of SLC26A4 were detected in 72 patients, including c.919-2A>G in 79.2%（57/72）. A novel deletion（c.1703_1707+6del） was discovered. Among 65 cases, truncated mutations were 89.2%（58/65）, 52.3%（34/65）, 28（43.1%） and 7（10.8%）. No significant differences were observed in the midpoint diameter of the vestibular aqueduct and the incidence of incomplete partitioning typeⅡ（IP-Ⅱ） of the cochlea among the three groups of patients. Moreover, there was no difference in the midpoint diameter of different vestibular pipes or the combination with IP-Ⅱ. Conclusion:The most common mutation site of SLC26A4 in EVA patients in Guizhou is c.919-2A>G, though genotype-phenotype correlations remain elusive. The detection of 27 mutation sites and the discovery of new mutation sites suggested the precise diagnostic significance of NGS technology in EVA patients in Guizhou.
Humans ; Sulfate Transporters ; Vestibular Aqueduct/abnormalities* ; Mutation ; Membrane Transport Proteins/genetics* ; Hearing Loss, Sensorineural/genetics* ; Male ; Female ; Child ; Adolescent ; Child, Preschool ; Adult ; Young Adult ; Phenotype ; High-Throughput Nucleotide Sequencing

Objective:To observe the therapeutic effect of compound Duzhong Jiangu granule on Kashin-Beck disease (KBD), and to provide a new alternative for the treatment of patients with KBD.Methods:According to the principle of random distribution, patients with KBD diagnosed clinically in Linyou County and Yongshou County of Shaanxi Province who meet the inclusion criteria were divided into a traditional Chinese medicine group and a Western medicine group, and they were treated with compound Duzhong Jiangu granule (Chinese medicine group) and ibuprofen sustained release capsule + 21 jinvita + chondroitin sulfate (Western medicine group), respectively, for a duration of one month. Questionnaire survey was conducted to collect the clinical data of all survey respondents before and after medication by using the Joint Dysfunction Index Scale and the Western Ontario and McMaster Universities Arthritis Index (WOMAC) scales, for evaluation and analysis. The incidence of adverse reactions of the two groups were recorded.Results:A total of 218 KBD patients that met the inclusion criteria were selected, including 167 patients in the Chinese medicine group and 51 patients in the Western medicine group. There were 94 males and 73 females in the Chinese medicine group, aged (62.93 ± 6.72) years. In the Western medicine group, there were 18 males and 33 females, aged (63.29 ± 7.02) years. There was no statistically significant difference in age between the two groups ( Z = - 0.24, P = 0.813). After taking the compound Duzhong Jiangu granules for treatment of KBD patients in the traditional Chinese medicine group, there were significant changes in the number of patients with joint rest pain, joint movement pain, morning stiffness, maximum walking distance and lower limb mobility in the Joint Dysfunction Scale compared to before treatment. The difference between before and after medication was statistically significant (χ 2 = 37.93, 29.64, 50.40, 13.57, 25.25, P < 0.001). After 1 month of medication, there were 13 cases of significant effect, 64 cases of effectiveness, and 90 cases of ineffectiveness in the traditional Chinese medicine group, with a total effective rate of 46.11%. There were 0 cases of significant improvement, 13 cases of effectiveness, and 38 cases of ineffectiveness in the Western medicine group, with a total effective rate of 25.49%. The difference in total effective rates between the two groups was statistically significant (χ 2 = 8.62, P = 0.013). In addition, there was a statistically significant difference in the improvement of lower limb mobility (difficulty of daily activities) between the Chinese medicine group and the Western medicine group (χ 2 = 8.21, P = 0.017). After taking medication, the joint pain, stiffness, and difficulty of daily activities scores in the WOMAC scale of KBD patients in the Chinese medicine group and the Western medicine group were significantly reduced. The differences in scores before and after medication were statistically significant (Chinese medicine group, Z = - 7.60, - 7.74, - 9.75, P < 0.001; Western medicine group, Z = - 5.20, - 3.81, - 3.93, P < 0.001). There was a significant differences in the improvement degree of daily activity difficulty and total score between the Chinese medicine group and the Western medicine group ( Z = - 3.75, - 3.34, P < 0.01). During the medication period, the incidence of adverse reactions in the traditional Chinese medicine group was lower than that in the western medicine group (Chinese medicine group, 29.34%; Western medicine group, 45.09%, χ 2 = 4.38, P = 0.036). Conclusions:Compound Duzhong Jiangu granule has a good therapeutic effects on KBD, significantly improving joint dysfunction of patients. It has advantages in improving the activity ability of KBD patients and reducing the difficulty of daily activities, and has less adverse reactions else.

Objective With the help of computerized tomography (CT) and susceptibility-weighted imaging (SWI) examinations, to analyze and compare the incidence and the prevalent sites of microbleeds (CMBs) in control group, diabetes group, hypertension group and diabetic hypertension group with intracerebral CMBs. The risk factors for the onset of CMBs also will be explored, in order to provide a basis for the selection of clinical diagnosis and treatment options for CMBs. Methods A collection of 174 non-acute patients in the Department of Neurology of Shandong Electric Power Central Hospital from May 2019 to October 2020, including 101 males and 73 females, aged 45-89 years, with an average age of 72 ± 5 years. Divided into four groups according to disease types: Hypertension with type 2 diabetes group, Hypertension group, Type 2 diabetes group, Control group (no hypertension and type 2 diabetes). All patients underwent CT and MRI SWI sequence scans. The 4 groups of patients were matched in terms of gender and age, excluding the influence of gender and age. The incidence, number and distribution of CMBs in the four groups were counted, and the CT and SWI imaging manifestations of CMBs in the four groups were analyzed and compared. Results CT did not find the incidence of CMBs in the brain of patients. SWI imaging showed that CMBs were detected in 58 of 174 patients (incidence rate 33.3%), and CMBs occurred in 24 cases of diabetes and hypertension group (incidence rate 57.1%). CMBs occurred in 21 cases in the hypertension group (incidence rate 46.7%), 8 cases in the diabetes group (incidence rate 22.2%), and CMBs occurred in 5 cases in the control group (incidence rate 9.8%). The incidence of CMBs in the hypertension group and the diabetes combined with hypertension group was significantly higher than that in the control group (P < 0.05). There was no significant difference in the incidence of CMBs in patients with simple diabetes compared with the control group. Also, there was no significant difference in the incidence of CMBs between the diabetes combined with hypertension group and the simple hypertension group. Conclusion CT did not find the incidence of CMBs in the brains of patients in the non-acute phase. SWI could clearly show the CMBs of the enrolled patients. Diabetes is less likely to cause an increase in CMBs. Hypertension is significantly related to the occurrence of CMBs. CMBs associated with hypertension or/and diabetes are common in the basal ganglia and thalamus, which is different from cerebral amyloid angiopathy, which provides a basis for the choice of clinical treatment options.

OBJECTIVE：To establish pre -column derivatization-HPLC fingerprint of Polyporus polysaccharide ，and to determine the contents of main monosaccharide components ，so as to provide reference for quality evaluation of Polyporus umbellatus. METHODS ：Polyporus polysaccharide was extracted with boiling water and precipitated by ethanol and deproteinized by Sevage from 11 batches of P. umbellatus from different producing areas. The samples were firstly hydrolyzed with trifluoro-acetic acid （TFA）and then derivatized by 1-phenyl-3-methyl-5-pyrazolone（PMP）. HPLC analysis was then conducted. The determination was carried out on HypersiL BDS C 18 column with mobile phase composed of 0.1 mol/L phosphate buffer （pH 6.84）-acetonitrile（84∶16，V/V）by gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm， and column temperature was 30 ℃. The sample size was 20 µL. The similarity of 11 batches of Polyporus polysaccharide was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System （2012A edition ），and the contents of main monosassharide components were detected. The peak was identified by comparing with the reference substance ，and cluster analysis was performed by using SPSS 23.0 software. RESULTS ：In HPLC fingerprints of the 11 batches of samples ，3 common peaks were identified ，namely mannose ，glucose and galactose. The similarity of all samples was above 0.94. Cluster analysis classified 11 batches of samples into three categories. S 1-S6，and S 8 were grouped into category 1；S7，S10 and S 11 were grouped into category 2；S9 was individually grouped into one category. Results of content determination showed that the contents of mannose ranged from 1.571 to 8.771 mg/g；those of glucose ranged from 26.072 to 132.194 mg/g，and those of galactose ranged from 3.420 to 36.593 mg/g. CONCLUSIONS ：Established pre-column derivatization HPLC fingerprints can provide reference for quality evaluation of P. umbellatus . The monosaccharide composition of different batches of Polyporus polysaccharide is the same ；there is no significant correlation between fingerprint characteristic peak and the origin of herbs ；there is significant difference in the content of monosaccharide of P. umbellatus .

OBJECTIVE To assess the association of single nucleotide polymorphisms (SNPs) of the T-cadherin (CDH13) gene with metabolic syndrome (MS) among ethnic Han Chinese.METHODS Genotypes of 6 SNPs(rs11646213, rs12596316, rs3865188, rs12444338, rs12051272, and rs7195409) of the CDH13 gene among 453 patients with MS and 526 controls were determined with a TaqMan method, and their association with MS was assessed. RESULTS For 5 SNPs (rs11646213, rs3865188, rs12444338, rs12051272, and rs7195409), no difference was found in allelic and genotypic frequencies of the CDH13 gene between the two groups. Comparing with rs12596316 (AA+GG) genotype, rs12596316 AG genotype has significantly increased the risk of MS(P = 0.01,OR = 1.38,95%CI: 1.07-1.78), though no association was found between particular alleles of the rs12596316 with MS.There was no difference in the frequencies of rs11646213-rs12596316-rs3865188-rs12444338-rs12051272 haplotype between the two groups(P > 0.05). CONCLUSION No association was found between the five SNPs (rs11646213, rs3865188, rs12444338, rs12051272 and rs7195409) of the CDH13 gene with the MS, while the rs12596316AG genotype of the CDH13 gene is associated with the susceptibility to MS among ethnic Han Chinese.

Background:Deoxyhypusine synthase(DHPS)is a key factor in post-translational modification of the precursor of eukaryotic initiation factor 5A(eIF-5A),and eIF-5A is closely related to the regulation of proliferation and invasion of tumor cells. Aims:To investigate the expression of DHPS in gastric cancer and its clinical significance,and to explore the possible mechanism of its effect on metastasis of gastric cancer. Methods:Tissue microarray containing 92 gastric cancer tissues and paired adjacent cancerous tissues was employed to detect the DHPS expression by using immunohistochemical staining,and the correlation of DHPS expression with the clinicopathological characteristics of gastric cancer was analyzed. DHPS-siRNA and GC7,an inhibitor of DHPS were used,respectively to intervene human gastric cancer cell line MGC803. The invasive ability of MGC803 cells was assessed by cell invasion assay,and the expressions of metastasis-related proteins including vascular endothelial growth factor(VEGF),matrix metalloproteinase 2(MMP2)and MMP9 were detected by ELISA. Results:In 62(67.4%)cases of gastric cancer,DHPS was highly expressed,and its expression was closely related to tumor diameter,TNM stage and depth of invasion(P <0.05),but not related to gender,age,lymph node metastasis and distant metastasis(P >0. 05). Both DHPS-siRNA and GC7 could down-regulate the invasiveness of MGC803 cells,while the former could also reduce the expressions of VEGF,MMP2 and MMP9 proteins(P <0.05). Conclusions:DHPS is highly expressed in gastric cancer and associated with tumor invasion and progression. DHPS is expected to be a new target for diagnosis and treatment of gastric cancer because of its regulatory effect on invasion and metastasis of tumor cells.

ObjectiveThe purpose of this study was to observe the correlation of mTOR and VEGF gene with nephropathy indicators in diabetic rats. Methods Forty-eight male Sprague-Dawley(SD)rats were divided into diabetes mellitus group(DM=28)and control group(NDM=20). Diabetic models were produced by injection of streptozotocin. In the courses of 12,16,20 and 24 weeks,the histology scores(HS)and glomerular basement membrane(GBM)thickness were collected. The protein and mRNA expressions of the gene of mTOR,VEGF and VEGFR2 were observed by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-Q-PCR)by SYBR Green. And the standardized cycle of threshold(SCt)was regarded as the indicators of the mRNA expression. Results HS and GBM thickness were significantly higher in DM rats than those in NDM rats,especially in DM rats of the courses of 20 and 24 weeks(P < 0.01). IHC scores of VEGF and VEGFR2 were higher in total DM rats and were positively correlated with each other. There were positive correlations between HS with VEGF and VEGFR2 in total DM rats(P < 0.05). SCts of VEGF and VEGFR2 were significantly higher and were positively correlated with each other in total DM rats(P < 0.01). SCt of VEGF and GBM thickness showed positive correlation in total DM rats. SCt of VEGF was highest in the course of 12w DM rats. SCt of VEGFR2 gradually decreased following by the diabetic course,and was lowest in the course of 24w. There were no significantly differences in IHC scores and SCt of mTOR between DM and NDM rats. But the IHC scores of mTOR,VEGF and VEGFR2 were positively correlated with each other and with HS in total DM rats(P < 0.05). Conclusion HS and GBM thickness were higher in diabetic rats,especially in the course of 24w,which could reflect the injury of nephropathy. The protein and mRNA of VEGF and VEGFR2 were high expressed in kidney of DM rats and increased with the increasing of diabetic course. The mRNA expression of VEGF was positively correlated with GBM thickness of in diabetic nephropathy(DN). The protein expressions of mTOR,VEGF and VEGFR2 might have synergistic effects in DN of DM rats. But the results could not exclude the influences of different courses,sample size and experimental conditions.

Objective To evaluate the effects of mild hypothermia performed at different times on the levels of glutamate, Bcl-2 and Bax during global cerebral ischemia-reperfsusion (I/R) in rats. Methods Twenty-four male SD rats weighing 230-270 g were randomly divided into 4 groups according to the time at which mild hypothermia was performed ( n =6 each): group A, B, C and D. Global cerebral ischemia was produced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and occlusion of bilateral common carotid arteries) in the 4 groups. In group B, C and D, the nasopharyngeal temperature was reduced to 32.5-33.5 ℃ and maintained for 1 h. Ischemia was performed after termination of cooling in group B. While ischemia was performed, cooling was started in group C. While reperfusion was performed, cooling was started in group D. Rewarming was started after termination of cooling in group B, C and D. Samples of dialysate in hippocampal CA1 area were collected every 10 min during 100 min reperfusion for determination of glutamate concentrations by high-performance capillary electrophoresis with laser-induced fluorescence detection ( HPCE-LIF). The brain tissues were taken at 3 h of reperfusion for determination of the expression of Bax and Bcl-2 in hippocampal CA1 area, and the ratio of Bcl-2/Bax was calculated. Results Compared with group A, the glutamate concentrations were significantly decreased, the Bcl-2 expression was up-regulated, the Bax expression was down-regulated and the ratio of Bcl-2/Bax was increased in the other thee groups ( P ＜ 0.05). Compared with group B, the Bcl-2 expression was significantly down-regulated, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group C ( P ＜0.05). The glutamate concentrations were significantly increased, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group D compared with group C ( P ＜ 0.05 ). Conclusion The earlier cooling is performed, the better the cerebral protective effect is during global cerebral I/R in rats.

Objective To set up the isolated perfused rat liver model. Methods Rat livers were harvested after the cannulation of the portal vein and bile duct. The reperfusion solution was Krebs-Henseleit solution containing bovine albumin serum and sodium taurocholate. The original CZ-1 isolated perfused rat liver system contained two subsystems: recirculating perfusion system and heat-exchange system. Then we modified the original CZ-1 system and omitted the heat-exchange system.The modified CZ-1 system consisted of a thermostatically regulated water bath,a peristaltic pump,a 4-neck round-bottom flask, a flow meter, an in-line manometer, a glass organ chamber, an iron support and a set of recirculating pipe line. Then the livers were connected via the portal vein to the modified CZ-1 system for 120 min. After 120 min reperfusion, bile production was evaluated. Routine HE staining and electron microscopic examination of hepatic tissues were also performed. Results The was not significantly different from that reported by references. Hepatic tissues in reperfusion group were also morphologically normal Conclusion The CZ-1 isolated perfused rat liver system was cost-effective and reliable to use. It was easy to run and is the ideal model for investigation of organ preservation solution.