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ObjectiveTo discuss the diagnostic methods of global developmental delay caused by 10q24.3 heterozygous loss. MethodsA retrospective analysis was conducted on the clinical data of one child with global developmental delay， and the results of low depth whole-genome copy number variation sequencing （CNVseq） and family whole exome sequencing （WES） of the child and his parents. ResultsThe patient was a 10-month-old male with developmental retardation in four areas， with some special features （ocular hypertelorism， strabismus， flat nose bridge， protruding forehead， cleft palate， high palatal arch， etc.） and hypotonia of limbs. The CNVseq and WES test showed that the patient had new 10q24.3 heterozygosis loss. Because this region contains the gene SUFU associated with basal cell nevus syndrome and the gene CNNM2 associated with hypomagnesemia， seizures， and mental retardation， and the gene TRIM8 associated of Focal segmental glomerulosclerosis with neurodevelopmental syndrome， we speculated that the cause of the disease in the child was highly related to the heterozygosity deletion of SUFU gene and CNNM2 gene and TRIM8 gene. ConclusionGenetic testing should be improved as soon as possible for children with global developmental delay and special facial manifestations， so as to make clear diagnosis and to judge prognosis.
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AIM: To observe the visual improvement and anatomical closure of patients with full-thickness macular hole(FTMH)accompanied by lamellar hole-associated epiretinal proliferation(LHEP)after vitrectomy.METHODS: A retrospective study of clinical cases. A total of 28 cases(28 eyes)of patients diagnosed with FTMH in our hospital between January 2018 and January 2022 were included, and they were divided into 12 cases(12 eyes)in the LHEP group and 16 cases(16 eyes)in the non-LHEP group according to the presence or absence of LHEP. All subjects had undergone vitrectomy. The best corrected visual acuity(BCVA)before and 1a after operation, hole closure, continuity of ellipsoid zone and external limiting membrane(ELM), intraocular pressure(IOP), and postoperative complications were compared between the two groups.RESULTS: Preoperative BCVA(LogMAR)was 0.80±0.17 in the LHEP group and 0.92±0.27 in the non-LHEP group(t=1.406, P=0.172); BCVA at 1a after operation was 0.54±0.14 in the LHEP group and 0.39±0.10 in the non-LHEP group(t=3.399, P=0.002). BCVA at 1a after operation was significantly improved in both groups compared with that before operation(tLHEP group=4.029; PLHEP group=0.001; tnon-LHEP group=7.445, Pnon-LHEP group=0.001); the difference in BCVA(LogMAR)before and after the operation was 0.27±0.16 in the LHEP group and 0.52±0.26 in the non-LHEP group(t=3.153, P=0.002). The hole closed in both groups 1a after the operation, and the closure rate was 100%. The ellipsoid zone closed in 20%(2/12)of patients in the LHEP group and 56%(9/16)in the non-LHEP group(Pellipsoid zone=0.04); the ELM closed in 25%(3/12)of patients in the LHEP group and 69%(11/16)in the non-LHEP group(PELM=0.027). A postoperative transient IOP elevation occurred in 2 eyes of the LHEP group and 3 eyes of the non-LHEP group, respectively. There were no significant cataract or serious complications after the vitrectomy in either group.CONCLUSIONS: Compared with FTMH patients without LHEP, the BCVA improvement after vitrectomy was less in FTMH patients with LHEP. Although their macular hole was closed, the closure rates of ellipsoid zone and ELM were lower, and the duration of continuity interruption was longer, so the prognosis requires further clarification.
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The glymphatic system plays a pivotal role in maintaining cerebral homeostasis. Chronic cerebral hypoperfusion, arising from small vessel disease or carotid stenosis, results in cerebrometabolic disturbances ultimately manifesting in white matter injury and cognitive dysfunction. However, whether the glymphatic system serves as a potential therapeutic target for white matter injury and cognitive decline during hypoperfusion remains unknown. Here, we established a mouse model of chronic cerebral hypoperfusion via bilateral common carotid artery stenosis. We found that the hypoperfusion model was associated with significant white matter injury and initial cognitive impairment in conjunction with impaired glymphatic system function. The glymphatic dysfunction was associated with altered cerebral perfusion and loss of aquaporin 4 polarization. Treatment of digoxin rescued changes in glymphatic transport, white matter structure, and cognitive function. Suppression of glymphatic functions by treatment with the AQP4 inhibitor TGN-020 abolished this protective effect of digoxin from hypoperfusion injury. Our research yields new insight into the relationship between hemodynamics, glymphatic transport, white matter injury, and cognitive changes after chronic cerebral hypoperfusion.
Subject(s)
Animals , Mice , Brain Ischemia , Carotid Stenosis/drug therapy , Cognitive Dysfunction/etiology , Digoxin , Disease Models, Animal , Mice, Inbred C57BL , White MatterABSTRACT
Ubiquitin-proteasome system (UPS) plays an important role in neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). The discovery of UPS activators for anti-neurodegenerative diseases is becoming increasingly important. In this study, we aimed to identify potential UPS activators using the high-throughput screening method with the high-content fluorescence imaging system and validate the neuroprotective effect in the cell models of AD. At first, stable YFP-CL1 HT22 cells were successfully constructed by transfecting the YFP-CL1 plasmid into HT22 cells, together with G418 screening. The degradation activity of the test compounds via UPS was monitored by detecting the YFP fluorescence intensity reflected by the ubiquitin-proteasome degradation signal CL1. By employing the high-content fluorescence imaging system, together with stable YFP-CL1 HT22 cells, the UPS activators were successfully screened from our established TCM library. The representative images were captured and analyzed, and quantification of the YFP fluorescence intensity was performed by flow cytometry. Then, the neuroprotective effect of the UPS activators was investigated in pEGFP-N1-APP (APP), pRK5-EGFP-Tau P301L (Tau P301L), or pRK5-EGFP-Tau (Tau) transiently transfected HT22 cells using fluorescence imaging, flow cytometry, and Western blot. In conclusion, our study established a high-content fluorescence imaging system coupled with stable YFP-CL1 HT22 cells for the high-throughput screening of the UPS activators. Three compounds, namely salvianolic acid A (SAA), salvianolic acid B (SAB), and ellagic acid (EA), were identified to significantly decrease YFP fluorescence intensity, which suggested that these three compounds are UPS activators. The identified UPS activators were demonstrated to clear AD-related proteins, including APP, Tau, and Tau P301L. Therefore, these findings provide a novel insight into the discovery and development of anti-AD drugs.
Subject(s)
Humans , Alzheimer Disease/drug therapy , Neuroprotective Agents , Optical Imaging , Proteasome Endopeptidase Complex , UbiquitinABSTRACT
Purpose@#Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen of coronavirus disease 2019. Diagnostic methods based on the clustered regularly interspaced short palindromic repeats (CRISPR) have been developed to detect SARSCoV-2 rapidly. Therefore, a systematic review and meta-analysis were performed to assess the diagnostic accuracy of CRISPR for detecting SARS-CoV-2 infection. @*Materials and Methods@#Studies published before August 2021 were retrieved from four databases, using the keywords “SARS-CoV-2” and “CRISPR.” Data were collected from these publications, and the sensitivity, specificity, negative likelihood ratio (NLR), positive likelihood ratio (PLR), and diagnostic odds ratio (DOR) were calculated. The summary receiver operating characteristic curve was plotted for analysis with MetaDiSc 1.4. The Stata 15.0 software was used to draw Deeks’ funnel plots to evaluate publication bias. @*Results@#We performed a pooled analysis of 38 independent studies shown in 30 publications. The reference standard was reverse transcription-quantitative PCR. The results indicated that the sensitivity of CRISPR-based methods for diagnosis was 0.94 (95% CI 0.93–0.95), the specificity was 0.98 (95% CI 0.97–0.99), the PLR was 34.03 (95% CI 20.81–55.66), the NLR was 0.08 (95% CI 0.06– 0.10), and the DOR was 575.74 (95% CI 382.36–866.95). The area under the curve was 0.9894. @*Conclusion@#Studies indicate that a diagnostic method based on CRISPR has high sensitivity and specificity. Therefore, this would be a potential diagnostic tool to improve the accuracy of SARS-CoV-2 detection.
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The high shear wet granulation(HSWG) process of Chinese medicine has a complicated mechanism. There are many influencing factors that contribute to this process. In order to summarize the manufacturability of different kinds of materials in HSWG, this paper constructed a material library composed of 11 materials, including 4 Chinese medicine extracts and 7 pharmaceutical excipients. Each material was described by 22 physical parameters. Several binders were employed, and their density, viscosity and surface tension were characterized. Combining empirical constraints and the principle of randomization, 21 designed experiments and 8 verification experiments were arranged. The partial least squares(PLS) algorithm was used to establish a process model in prediction of the median granule size based on properties of raw materials and binders, and process parameters. The surface tension and density of binders, as well as the maximum pore saturation were identified as key variables. In the latent variable space of the HSWG process model, all materials could be divided into three categories, namely the Chinese medicine extracts, the diluents and the disintegrants. The granulation of Chinese medicine extracts required low viscosity and low amount of binder, and the resulted granule sizes were small. The diluent powders occupied a large physical space, and could be made into granules with different granule sizes by adjusting the properties of binders. The disintegrants tended to be made into large granules under the condition of aqueous binder. The combination use of material database and multivariate modeling method is conducive to innovate the knowledge discovery of the wet granulation process of Chinese medicine, and provides a basis for the formulation and process design based on material attributes.
Subject(s)
Drug Compounding , Excipients , Medicine, Chinese Traditional , Particle Size , Powders , Tablets , Technology, PharmaceuticalABSTRACT
OBJECTIVE@#To measure the three-dimensional morphology of the labial supraosseous gingiva (SOG) and the thickness of related labial bone in maxillary anterior teeth of periodontally healthy Han nationality youth using soft tissue indirect imaging cone-beam computed tomography (CBCT).@*METHODS@#Twenty-five periodontally healthy subjects (11 males and 14 females) with 150 maxillary anterior teeth were involved in this study. A special impression with radiopaque material including the maxillary teeth was made, then a CBCT scan with the elastomeric matrix in position was taken for each subject. The imaging data were generated and transferred to a volumetric imaging software in which three-dimensional reconstruction was conducted and the image analyses were carried out. Measurements were made at the site of labial center of the maxillary anterior teeth. The height of the SOG, the distance between cemento-enamel junction (CEJ) and bone crest, the gingival thickness at the CEJ, and the thickness of bone 2 mm below the labial bone crest were measured and the correlation analysis between the parameters was made. All the data analyses were performed using SPSS 22.0. The data were analyzed with ANVOA and Pearson correlation tests with the significance level at α=0.05.@*RESULTS@#The mean SOG values were (3.49±0.70) mm, (3.48±0.81) mm, and (3.54±0.67) mm for central incisors, lateral incisors and canines, respectively. There were no statistically significant differences among the different sites (P > 0.05). The mean gingival thickness values were (1.45±0.23) mm, (1.13±0.24) mm, (1.14±0.22) mm for central incisors, lateral incisors and canines, respectively. The gingival thickness of the central incisors was the largest among the maxillary anterior teeth with statistically significant difference (P < 0.05). No correlation was found between the SOG and gingival thickness among the maxillary anterior teeth (P > 0.05).@*CONCLUSION@#The gingival thickness of central incisors was the largest and the supraosseous gingival height had no correlation with gingival thickness among the periodontally healthy maxillary anterior teeth.
Subject(s)
Adolescent , Female , Humans , Male , Cone-Beam Computed Tomography , Gingiva/diagnostic imaging , Incisor/diagnostic imaging , Maxilla/diagnostic imaging , Tooth CervixABSTRACT
According to the commonly used tablet compressibility, compactability and tabletability equation, the influence of pressure range on the fitting results and parameters of different compression equations was studied, and the optimal pressure range of different equations was determined. Plastic material microcrystalline cellulose (MCC) PH102, brittle material spray dried lactose and Chinese medicine Sanqi were used as experimental objects, the compression curves of tablets were obtained by the combination of dies with different diameters. For Heckel equation, the shape of Heckel section of different materials is not uniform, and the specified linear fitting range cannot be obtained, therefore, different distances between fitting pressure starting point and starting point were set to observe the influence of pressure range on R2 of Heckel equation. The Kawakita equation, Gurnham equation, Ryshkewitch-Duckworth (R-D) equation and Power equation are fitted in three different pressure ranges of 15-200, 15-300 and 15-400 MPa, respectively. In order to find the best linear region of Heckel equation, the 3D scatter diagram of "starting point of pressure, pressure range and R2" is drawn. The best linear pressure ranges of Heckel curves of MCC, lactose and Sanqi were 20-170, 20-220 and 10-90 MPa, respectively. It is proved that the 3D scatter diagram is an effective method to find the linear range of Heckel equation. The change of pressure range has little influence on the curve fitting effect and compression parameters of Kawakita equation, Gurnham equation and Ryshkewitch-Duckworth equation. The low pressure range of 15-200 MPa can meet the fitting requirements of Kawakita equation, Gurnham equation, R-D equation and Power equation for different materials. Therefore, only by optimizing the pressure range, can the good fitting effect be ensured and the obtained compression parameters be more reliable and interpretable.
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Objective:To investigate the influence of particle size on density of binary powder mixture of traditional Chinese medicine （TCM）， and to provide reference for formulation design of TCM preparations. Method:Three groups of binary powder mixtures with different particle size ratio （<italic>α</italic>） were constructed， namely Oroxyli Semen-microcrystalline cellulose PH102 （MCC PH102） （<italic>α</italic>=0.071 7）， Stellariae Radix-MCC PH200 （<italic>α</italic>=0.158 7） and Angelicae Sinensis Radix-MCC KG802 （<italic>α</italic>=0.840 6）. Binary powder mixtures with nine mass ratios （90∶10， 80∶20， 70∶30， 60∶40， 50∶50， 40∶60， 30∶70， 20∶80 and 10∶90） were prepared for each group， and 27 binary powder mixtures containing TCM were obtained. The particle size distribution， density and other parameters of six single materials and 27 binary powder mixtures were characterized. Based on the packing theory and multivariate analysis， the effects of particle size related parameters on the filling structure and density of the binary powder mixtures were elucidated. Result:The <italic>α</italic> of Angelicae Sinensis Radix-MCC KG802 binary mixture system was larger than the replacement rate （<italic>α</italic><sub>r</sub>=0.741 0）， and its density had a good linear relationship with the mass ratio， which conformed to the replacement mechanism. The <italic>α</italic> of Oroxyli Semen-MCC PH102 binary mixture system was smaller than the critical ratio （<italic>α</italic><sub>c</sub>=0.154 0）， and its density was nonlinear with the mass ratio of components， which conformed to the filling mechanism. The <italic>α</italic> of Stellariae Radix-MCC PH200 binary mixture system was between <italic>α</italic><sub>c</sub> and <italic>α</italic><sub>r</sub>， its density was affected by both of replacement mechanism and filling mechanism. Based on the partial least squares （PLS） model， the variable importance in the projection （VIP） analysis further proved that the mixing mass ratio （VIP value=1.62）， <italic>α</italic> （VIP value=1.13） and <italic>D</italic><sub>10</sub> （the corresponding particle size when the particle size distribution accumulated to 10%， VIP value=1.06） were the key factors affecting the density of binary powder mixtures of TCM. Conclusion:In the binary powder mixtures of TCM， the linearity relationship between density and mass ratio is largely depended on particle size difference of components.
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A 90-year-old man was diagnosed with primary gastric diffuse large B-cell lymphoma (PGDLBL) by PET/CT examination, gastroscopy, biopsy and histopathological analysis at a regular physical check in April, 2016. The patient received R-CO chemotherapy (rituximab, cyclophosphamide, and vincristine) and radiotherapy subsequently, with enteral nutritional treatment through 3-cavity nasogastric tube due to development of pyloric obstruction. To satisfy patient's strong desire of eating by himself, we performed surgery of exploratory laparotomy and Roux-en-Y gastric bypass (RGB) to relieve pylorus obstruction. Postoperatively, the patient resumed oral feeding, supplemented by nasogastric tube feeding at 1350 - 1550 Kcal daily. He is now 94 years old with fairly well nutrition and normal communication. The outcome of 4 year follow-up suggests that nutritional treatment and palliative medicine are important for improving prognosis and life-quality of very elderly patients with end-stage tumors apart from the effective chemotherapy, radiotherapy, and surgery.
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Objective:To investigate the influence of glucose regulatory protein 78 (GRP78) on prognosis and tumor cell proliferation of hepatocellular carcinoma.Methods:The experimental study and retrospective cohort study were conducted. Based on hepatocellular carcinoma tissue chip, in vitro culture of Huh7 and Hep3B hepatoma cells and LO2 normal hepatic cell, and combined with immunohistochemical staining, cell transfection, quantitative real-time polymerase chain reaction (qRT-PCR), Western blot detection, cell proliferation experiments, cell clone formation experiments and high-throughput transcription histological analysis, the GRP78 expression in hepatoma cells was analyzed. Huh7 and Hep3B hepatoma cells being transfected with the GRP78 gene-specific shRNA lentiviruses or the negative control shRNA lentivirus were set as the GRP78 gene-specific shRNA lentivirus group and the negative control shRNA lentivirus group respectively. Observation indicators: (1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients; (2) analysis of factors affecting the prognosis of hepatocellular carcinoma patients; (3) effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells; (4) effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells; (5) effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells. Measurement data of the normal distribution were expressed as Mean± SD, and comparison of groups was conducted using the t test or ANOVA. Repeated measurement data were analyzed using repeated ANOVA. Count data were expressed as absolute numbers, and comparisons between groups was conducted using the chi-square test. COX proportional hazards regression model was used for univariate and multivariate analysis. The Kaplan-Meier method was used to calculate the survival time and draw survival curve, and the Log-rank test was used for generative analysis. Results:(1) GRP78 expression in hepatocellular carcinoma tissue and adjacent tissue and its correlation with the clinicopathological characteristics of hepatocellular carcinoma patients: results of immunohistochemical staining of hepatocellular carcinoma tissue chip showed that GRP78 was low-expressed in 53 cases and high-expressed in 37 cases of the 90 hepatocellular carcinoma tissues. GRP78 was low-expressed in 84 cases and high-expressed in 6 cases of the 90 paracancerous tissues. There was a significant difference in GRP78 expression between hepatocellular carcinoma tissues and paracancerous tissues ( P<0.05). (2) Analysis of factors affecting the prognosis of hepatocellular carcinoma patients: all 90 patients were followed up for 5 to 56 months, with a median follow-up time of 49 months. The median overall survival time and median disease progression-free survival time were 56 months and 53 months in the 53 hepatocellular carcinoma patients with GRP78 as low-expressed, versus 32 months and 19 months in the 37 hepatocellular carcinoma patients with GRP78 as high-expressed, respec-tively, showing significant differences ( χ2=17.482, 12.097, P<0.05). Results of univariate analysis showed that alanine aminotransferase (ALT), tumor pathological grading and GRP78 expression were related factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.317, 2.039, 3.740 and 2.194, 2.177, 2.927, 95% confidence interval as 1.150?4.671, 1.201?3.462, 2.116?6.612 and 1.048?4.593, 1.093?4.336, 1.492?5.742, P<0.05). Results of multivariate analysis showed that ALT >40 U/L, tumor pathological grading as Ⅲ-Ⅳ grade and GRP78 as high-expressed were independent risk factors affecting the 3-year overall survival rate and disease progression-free survival rate of hepatocellular carcinoma patients ( hazard ratio=2.438, 2.245, 3.223 and 3.046, 2.473, 3.307, 95% confidence interval as 1.114?5.334, 1.047?4.814, 1.396?7.440 and 1.337?6.940, 1.141?5.360, 1.399?7.819, P<0.05). (3) Effects of inhibiting of GRP78 expression on the proliferation of hepatoma cells: ①results of qRT-PCR showed that the relative expression of GRP78 messenger RNA (mRNA) in Huh7, Hep3B, and LO2 cells were 3.06±0.33, 4.42±0.60 and 1.00±0.02. There were significant differences in GRP78 mRNA expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=6.19, 5.42, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78 protein in Huh7, Hep3B, and LO2 cells were 1.65±0.01, 1.77±0.01 and 0.99±0.02. There were significant differences in GRP78 protein expression between Huh7 and LO2 cells or Hep3B and LO2 cells ( t=75.09, 108.10, P<0.05). ③Results of cell proliferation experiments showed that the growth rates in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 111.51%±0.35%, 144.85%±0.68%, 188.71%±3.62%, 282.51%±5.25% and 190.08%±0.58%, 285.76%±2.69%, 459.51%±4.29%, 597.88%±12.25%, showing signifi-cant differences ( Fgroups=1 360.000, Ftime=668.500, Finteraction=197.600, P<0.05). The growth rates in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells at 24, 48, 72 and 96 hours were 124.47%±0.25%, 153.25%±1.25%, 195.45%±3.19%, 282.51%±10.76% and 179.69%±0.33%, 322.67%±2.46%, 486.27%±5.82%, 622.35%±12.58%, showing significant differences ( Fgroups=1 222.000, Ftime=706.200, Finteraction=179.600, P<0.05). ④Results of the cell clone formation experiments showed that the number of cells in Hu7 GRP78 gene-specific shRNA lentiviruses group cells and Hu7 negative control shRNA lentivirus group cells were 125±3 and 435±17, showing a significant difference ( t=17.86, P<0.05). The number of cells in Hep3B GRP78 gene-specific shRNA lentiviruses group cells and Hep3B negative control shRNA lentivirus group cells were 138±3 and 388±7, showing a significant difference ( t=32.29, P<0.05). (4) Effects of inhibiting of GRP78 expression on the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of high-throughput transcription histological analysis showed that the relative expression rates of p53, p21, CDK2, CDK4, and CDK6 were 19%, 334%, 398%, 41% and 49% in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells comparing to the Hu7 negative control shRNA lentivirus group cells. ①Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.17±0.03, 4.05±0.71, 3.73±0.47, 0.49±0.09, 0.48±0.06, 0.36±0.07 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.05, 1.03±0.17, 1.00±0.07, 1.01±0.09, 1.02±0.14, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=14.62, 4.17, 5.72, 4.26, 3.49, 8.82, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.11±0.01, 4.28±0.43, 4.19±0.22, 0.44±0.01, 0.25±0.03, 0.68±0.04 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.09, 1.02±0.15, 1.00±0.06, 1.01±0.09, 1.01±0.08, 1.15±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.19, 7.14, 13.79, 6.37, 9.42, 9.61, P<0.05). ②Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.45±0.01, 1.98±0.05, 2.31±0.12, 0.75±0.03, 0.69±0.04, 0.82±0.03 in the Hu7 GRP78 gene-specific shRNA lentiviruses group cells, versus 1.01±0.05, 1.03±0.01, 1.00±0.02, 1.00±0.01, 1.01±0.02, 1.00±0.03 in the Hu7 negative control shRNA lentivirus group cells, showing significant differences ( t=11.07, 14.56, 11.30, 11.29, 10.55, 11.37, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.61±0.03, 1.98±0.16, 2.55±0.12, 0.85±0.03, 0.78±0.01, 0.54±0.02 in Hep3B GRP78 gene-specific shRNA lentiviruses group cells, versus 1.00±0.03, 1.05±0.02, 1.05±0.01, 1.05±0.02, 1.00±0.02, 1.00±0.02 in Hep3B negative control shRNA lentivirus group cells, showing significant differences ( t=10.97, 13.40, 12.35, 11.06, 12.45, 13.78, P<0.05). (5) Effects of HA15 on the proliferation and the gene and protein expression of p53, p21, CDK2, CDK4, and CDK6 in hepatoma cells: results of 50% inhibiting concentration (IC50) test of HA15 showed that the IC50 of HA15 for Huh7 and Hep3B cells at 48 hours were 9.98 μmol/L and 13.70 μmol/L. ①Huh7 and Hep3B cells were treated with 9.98 μmol/L and 13.70 μmol/L of HA15. Results of cell proliferation experiments showed that the growth rates at 24, 48, 72, and 96 hours were 112.81%±0.27%, 154.71%±1.45%, 237.66%±16.77%, 294.40%±14.92% in the HA15-Huh7 cells, versus 133.67%±0.49%, 352.93%±2.31%, 557.17%±4.89%, 662.60%±13.31% in the normal Huh7 cells, showing a significant difference ( Fgroups=766.800, Ftime=518.200, Finteraction=133.300, P<0.05). The growth rates at 24, 48, 72, and 96 hours were 121.27%±2.32%, 203.85%±3.18%, 240.80%±3.02%, 286.50%±7.10% in the HA15-Hep3B cells, versus 239.14%±1.02%, 362.00%±5.44%, 539.37%±10.80%, 694.79%±17.13% in the normal Hep3B cells, showing a signifi-cant difference ( Fgroups=594.300, Ftime=317.900, Finteraction=78.600, P<0.05). ②Results of qRT-PCR showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.27±0.05, 3.64±0.28, 4.13±0.41, 0.51±0.07, 0.39±0.03, 0.17±0.02 in the HA15-Huh7 cells, versus 1.02±0.14, 1.00±0.03, 1.00±0.05, 1.01±0.08, 1.01±0.09, 1.03±0.17 in the normal Huh7 cells, showing significant differences ( t=5.00, 9.25, 7.63, 4.73, 6.82, 5.01, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 mRNA were 0.28±0.03, 3.49±0.78, 4.31±0.53, 0.38±0.05, 0.36±0.04, 0.24±0.03 in the HA15-Hep3B cells, versus 1.01±0.11, 1.03±0.18, 1.01±0.08, 1.00±0.06, 1.02±0.15, 1.00±0.06 in the normal Hep3B cells, showing significant differences ( t=6.26, 3.08, 6.21, 7.97, 4.26, 11.08, P<0.05). ③Results of Western Blot detection showed that the relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.52±0.05, 1.94±0.08, 1.58±0.02, 0.89±0.00, 0.86±0.02, 0.74±0.01 in the HA15-Huh7 cells, versus 1.02±0.03, 1.00±0.03, 1.02±0.02, 1.04±0.03, 1.00±0.01, 1.01±0.02 in the normal Huh7 cells, showing significant differences ( t=11.54, 10.28, 11.03, 12.81, 13.67, 10.09, P<0.05). The relative expression of GRP78, p53, p21, CDK2, CDK4, and CDK6 protein were 0.57±0.02, 1.67±0.04, 1.41±0.04, 0.82±0.03, 0.70±0.02, 0.74±0.01 in the HA15-Hep3B cells, versus 1.03±0.01, 0.98±0.03, 1.00±0.03, 1.03±0.03, 1.01±0.01, 1.04±0.01 in the normal Huh7 cells, showing significant differences ( t=10.81, 11.54, 12.26, 13.62, 14.23, 10.17, P<0.05). Conclusions:High expression of GRP78 is an independent risk factor affecting the overall survival and disease progression-free survival of hepatocellular carcinoma patients. Inhibiting of GRP78 expression can reduce cell proliferation and the expression of p53, p21, CDK2, CDK4, and CDK6 mRNA and proteins in hepatoma cells.
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OBJECTIVE@#To evaluate the short-term outcome of regenerative surgery for peri-implantitis therapy.@*METHODS@#From March 2018 to January 2019, 9 patients with 10 implants who suffered from peri-implantitis were included in the present research. Vertical bone defect at least 3mm in depth with 2 or more residual bone walls was confirmed around each implant by radiographic examination. Restorations were replaced by healing abutments on 3 implants with the consent of the patients. Guided bone regeneration surgery was performed after a hygienic phase. During surgery, full thickness flaps were elevated on both buccal and lingual aspects. Titanium curette was used for inflammatory granulation tissue removal and implant surface cleaning. The implant surface was decontaminated by chemical rinsing with 3% hydrogen peroxide solution. After being thoroughly rinsed with saline, the bone substitutes were placed in bone defects which were covered by collagen membranes. 6 months after non-submerged healing, the clinical parameters including peri-implant probing depth (PD, distance between pocket bottom and peri-implant soft tissue margin) and radiographic bone level (BL, distance form implant shoulder to the first bone-to-implant contact) were used to evaluate the regenerative outcome. PD was measured at six sites (mesial, middle and distal sites at both buccal and lingual aspects) around each implant, and BL was measured at the mesial and distal surfaces of each implant on a periapical radiograph.@*RESULTS@#The deepest PD and largest BL of each implant ranged from 6-10 mm and 3.2-8.3 mm respectively. All the implants healed uneventfully after surgery. The mean peri-implant PD at baseline and 6 months after surgery were (6.2±1.4) mm and (3.1±0.6) mm respectively, and a mean (3.0±1.5) mm radiographic bone gain was observed, P<0.01. Treatment success was defined as: no sites with residual PD≥6 mm, no bleeding on probing, and BL elevation of at least 1 mm. Nine implants from 8 patients fulfilled the success criteria. Residual pockets with 6 mm in depth and bleeding on probing could be detected in only one implant.@*CONCLUSION@#Within the limitation of the present research, guided bone regeneration surgery can be used for the treatment of bone defect that resulted from peri-implantitis. Significant PD reduction and radiographic bone gain can be obtained after 6 months observation.
Subject(s)
Humans , Alveolar Bone Loss , Bone Substitutes , Collagen , Dental Implants , Peri-Implantitis , Surgical Flaps , Treatment OutcomeABSTRACT
Objective: To evaluate the incidence of cardiovascular disease (CVD), ischemic and hemorrhagic cardiovascular events among Chinese diabetic patients aged 40 years and above with different CVD risk levels. Methods: This study enrolled participants aged 40 years and above in 15 provinces from a prospective cohort study, the China-PAR project (Prediction for Atherosclerotic Cardiovascular Disease Risk in China). Participants were categorized into two groups according to the presence or absence of diabetes at baseline. Individuals were further classified into low (0-4.9%), moderate (5%-9.9%) and high risk groups (≥10%), based on predicted ten-year CVD risk using the China-PAR equations. Two followed-up surveys were conducted between 2007 and 2015 to identify CVD events, which were defined as nonfatal acute myocardial infarction, or death due to coronary heart disease, or stroke. Ischemic cardiovascular events included nonfatal acute myocardial infarction, or death due to coronary heart disease, or ischemic stroke. Hemorrhagic cardiovascular events included subarachnoid hemorrhage and intracerebral hemorrhage. The incidences of CVD, ischemic and hemorrhagic cardiovascular events were compared in diabetes and non-diabetes population with different CVD risk levels. Results: This study included 89 209 participants aged 40 years and above, the average follow-up period was 8.5 years. The age was (54.8±9.4) years, and 36 794 (41.2%) were men, and 5 730 (6.4%) were diabetic patients. In diabetes patients aged 40 years and above, 53.7% (3 075/5 730) were at high risk of CVD. Age-and sex-adjusted incidence of CVD, ischemic and hemorrhagic cardiovascular events (1 066.93/100 000 person-years, 824.23/100 000 person-years, and 211.56/100 000 person-years) were significantly lower in diabetes patients than those in non-diabetes population with high CVD risk (1 773.73/100 000 person-years, 1 228.18/100 000 person-years, and 446.49/100 000 person-years) (all P<0.001). Among high CVD risk populations, incidence of ischemic events was significantly higher in diabetic patients than in non-diabetes population (1 638.47/100 000 person-years vs. 1 228.18/100 000 person-years, P<0.001), but incidence of hemorrhagic events tended to be lower in diabetic patients than in non-diabetes population (415.70/100 000 person-years vs. 446.49/100 000 person-years, P=0.635). Incidence of ischemic and hemorrhagic events were similar between diabetes patients and non-diabetes population at low or moderate CVD risk groups (all P>0.05). Conclusions: More than half of diabetes patients aged 40 years and above in China have high CVD risk. The incidence of CVD, ischemic and hemorrhagic cardiovascular events are different in diabetic patients with different CVD risk levels.
Subject(s)
Adult , Humans , Male , Cardiovascular Diseases/epidemiology , China/epidemiology , Diabetes Mellitus/epidemiology , Incidence , Prospective Studies , Risk FactorsABSTRACT
Objective:To study the application of objective structured clinical examination (OSCE) based on organ system in the practice teaching for clinical interns.Methods:Undergraduate interns majored in clinical medicine in grade 2017 (control group) and 2018 (experimental group) of Jiangjin Central Hospital of Chongqing were selected as the research objects and divided into control group and experimental group. In the control group, the interns received traditional practice teaching, and the interns in the experimental group received traditional bedside teaching combined with OSCE based on organ system. Results of theoretical examination and OSCE test were used to evaluate the teaching effect, and feedbacks were obtained from teachers and students through questionnaire survey and field discussion, so as to compare the differences between the two teaching methods in cultivating clinical ability of medical students.Results:The average scores of theoretical examination of the two groups were similar, the difference was not statistically significant ( P>0.05). In OSCE test, the average scores and total scores of medical history collection station, physical examination station, skill operation station, doctor-patient communication station in the research group were significantly higher than those in the control group ( P<0.05). The average scores of the two groups in the auxiliary examination interpretation station and literature retrieval station were similar, and the differences were not statistically significant ( P>0.05). Conclusion:The OSCE based on organ system is obviously superior to traditional practice teaching in the development of medical students' comprehensive clinical capacity such as medical history collection, physical examination, skill operation and doctor-patient communication.
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Objective:To invetigate the influencing factors and clinical significance of liver function damage (LFD) in patients diagnosed with Corona Virus Disease 2019 (COVID-19).Methods:The retrospective case-control study was conducted. The clinicopathological data of 51 patients with COVID-19 who were admitted to the Sino-French New City Branch of Tongji Hospital Affiliated to Huazhong University of Science and Technology by the 5th group assisting team from the First Hospital of Jilin University from February 9th to 27th in 2020 were collected. There were 27 males and 24 females, aged from 36 to 86 years, with an average age of 68 years. The treatment modality was according to the diagnostic and therapeutic guideline for COVID-19 (Trial 6th edition) issued by National Health Commission. Observation indicators: (1) clinical data of patients; (2) analysis of liver function index and treatment of LFD; (3) analysis of influencing factors for LFD. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were described as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. The Logistic regression method was used for univariate analysis. Results:(1) Clinical data of patients: of the 51 patients, 21 were classified as ordinary type of COVID-19, 19 as severe type and 11 as critical type. In terms of medical history, 31 patients suffered from more than or equal to one kind of chronic disease, 20 had no history of chronic disease. Thirteen patients had the drinking history and 38 had no drinking history. Seven patients were hepatitis positive and 44 were hepatitis negative. Five patients had septic shock at admission, 5 had systemic inflammatory response syndrome (SIRS), and 41 had neither shock nor SIRS. The body mass index (BMI), time from onset to admission, temperature, heart rate, respiratory rate of the 51 patients were (24±3)kg/m 2, (13±5)days, 36.5 ℃ (range, 36.0-38.1 ℃), 82 times/minutes (range, 50-133 times/minutes), 20 times/minutes (range, 12-40 times/minutes). The white blood cell count, level of creatinine, and level of b-type natriuretic peptide within 24 hours after admission were 6.3×10 9/L [range, (2.2-21.7)×10 9/L], 75 μmol/L (range, 44-342 μmol/L), 214 ng/L (range, 5-32 407 ng/L). (2) Analysis of liver function index and treatment of LFD: the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), direct bilirubin (DBil), indirect bilirubin (IBil), activated partial thromboplastin time (APTT) and prothrombin time (PT) were 31 U/L (range, 7-421 U/L), 29 U/L (range, 15-783 U/L), 36 U/L (range, 13-936 U/L), 76 U/L (range, 41-321 U/L), 4.9 μmol/L (range, 2.6-14.3 μmol/L), 5.8 μmol/L (range, 2.6-23.9 μmol/L), 37.2 s (range, 30.9-77.1 s), 13.9 s (range, 12.5-26.7 s), respectively. The percentages of cases with abnormal ALT, AST, GGT, ALP, DBil, IBil, APTT and PT were 47.1%(24/51), 47.1%(24/51), 35.3%(18/51), 13.7%(7/51), 7.8%(4/51), 2.0%(1/51), 21.6%(11/51), and 19.6%(10/51), respectively. Of the 51 patients, LFD was detected in 10 patients classified as ordinary type, in 9 patients as severe type, and in 10 as critical type, respectively. In the 51 patients, 1 of 22 patients with normal liver function developed respiratory failure and received mechanical ventilation within 24 hours after admission, while 9 of 29 patients with abnormal liver function developed respiratory failure and received mechanical ventilation, showing a significant difference between the two groups ( χ2=5.57, P<0.05). (3) Analysis of influencing factors for LFD. Results of univariate analysis showed that clinical classification of COVID-19 as critical type was a related factor for LFD of patients ( odds ratio=10.000, 95% confidence interval: 1.050-95.231, P<0.05). Conclusions:COVID-19 patients with LFD are more susceptible to develop respiratory failure. The clinical classification of COVID-19 as critical type is a related factor for LFD of patients.
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OBJECTIVE@#To investigate the effect and mechanism of miR-124-3p-targeing regulating ABCA2 on chronic myelogenous leukemia cell K562-R.@*METHODS@#CML cells with miR-124-3p-overexpression and ABCA2-over-expression as well as subcutaneoustrans planted tumor nude mice were used as study objects. And the CML cells were divided into four groups: K562-R blank control, miR-124-3p mimic control, ABCA2-overexpression and mimic+PC ABCA2. The effects of miR-124-3p and ABCA2 on CML cells were analyzed. The levels of proliferation-, apoptosis- and autophagy- related protein were determined by Western blot. qRT-PCR was employed to detect the levels of miR-124-3p and ABCA2 in K562-R cells. The relationship between miR-124-3p and ABCA2 was validated by luciferase reporter system assays and bioinformatics. Hoechst/immunohistochemical staining and CCK-8 assay were performed to investigate the function involved.@*RESULTS@#miR-124-3p highly expressed in K562-S cells and lowly expressed in K562-R cells, however, ABCA2 lowly expressed in K562-S cells and highly expressed in K562-R cells. Over-expression of miR-124-3p significantly decreased ABCA2 level and cell growth, but increased autophagy and apoptosis in K562-R cells (P＜0.01). When ABCA2 was over-expressed, the K562-R cell growth was promoted and autophagy and apoptosis were inhibited (P＜0.01). The miR-124-3p promoted cell autophagy and apoptosis but inhibited cell growth in nude mice transplant tumor model (P＜0.01).@*CONCLUSION@#miR-124-3p can target ABCA2 to inhibit the growth of CML cells and promote the cell autophagy and apoptosis of CML cells.
Subject(s)
Animals , Humans , Mice , ATP-Binding Cassette Transporters , Apoptosis , Cell Proliferation , Imatinib Mesylate , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Mice, Nude , MicroRNAsABSTRACT
BACKGROUND@#The association of milk intake with cardiovascular disease (CVD) and cause-specific mortality remained controversial and evidence among the Chinese population was limited. We aimed to study the relationship between milk intake and CVDs among general Chinese adults.@*METHODS@#A total of 104,957 participants received questionnaire survey. Results of physical examination such as anthropometric measurements and biochemical tests during 2007 to 2008, demographic data and their information on milk intake were collected through standardized questionnaires. Cox proportional hazard regression models were used to calculate hazard ratios (HRs) and their corresponding 95% confidence intervals (CIs) of CVD incidence, cause-specific mortality and all-cause mortality related to milk intake. Restricted cubic splines (RCSs) were applied to examine dose-response associations.@*RESULTS@#Among the 91,757 participants with a median follow-up period of 5.8 years, we documented 3877 CVD cases and 4091 all-cause deaths. Compared with participants who never consumed milk, the multivariate-adjusted HRs (95% CIs) of CVD incidence for 1 to 150 g/day, 151 to 299 g/day, and ≥300 g/day were 0.94 (0.86-1.03) (P > 0.05), 0.77 (0.66-0.89) (P < 0.05), and 0.59 (0.40-0.89) (P < 0.05), respectively; each 100 g increase of daily milk intake was associated with 11% lower risk of CVD incidence (HR, 0.89; 95% CI: 0.85-0.94; P < 0.001), and 11% lower risk of CVD mortality (HR, 0.89; 95% CI: 0.82-0.97; P = 0.008) after adjustment for age, sex, residential area, geographic region, education level, family history of CVD, smoking, alcohol drinking, physical activity level, body mass index, and healthy diet status (ideal or not). RCS analyses also showed a linear dose-response relationship with CVD (P for overall significance of the curve <0.001; P for non-linearity = 0.979; P for linearity <0.001) and stroke (P for overall significance of the curve = 0.010; P for non-linearity = 0.998; P for linearity = 0.002) incidence, and CVD mortality (P for overall significance of the curve = 0.045; P for non-linearity = 0.768; P for linearity = 0.014) within the current range of daily milk intake.@*CONCLUSIONS@#Daily milk intake was associated with lower risk of CVD incidence and mortality in a linear inverse relationship. The findings provide new evidence for dietary recommendations in CVD prevention among Chinese adults and people with similar dietary pattern in other countries.
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OBJECTIVE@#To investigate the clinical effects of dual mobility total hip prosthesis in treating femoral neck fracture patients with hemiplegia.@*METHODS@#A retrospective analysis was performed on 18 patients with femoral neck fracture combined with hemiplegia who underwent dual mobility total hip prosthesis replacement from March 2014 to December 2016. The follow up data of these patients was complete. There were 5 males and 13 females, aged 65 to 70 years old with an average of (66.50±1.38) years. The left side was involved in 12 cases, while the right side in 6 cases. There were 4 cases with Garden Ⅲ type and 14 cases with type Ⅳ. Limb muscle strength of hemiplegia were in grade Ⅳ. The posterior-lateral approach of hip joint was used in surgery for all patients. The implant position, dislocation and loosening of the prosthesis were evaluated by X-ray examination. Harris hip score and the Merle D'aubigne score were used to assess the hip function in the follow up.@*RESULTS@#The operation duration was for 70-90 (81.56±7.48) min and the blood loss during the operation was for 160-200 (170.32± 12.56) ml. No blood was transfused during the operation. Postoperative incisions were healed at the first stage. The follow-up time was for 28-60(36.0±3.5) months. Harris hip score increased from 16.94±0.73 preoperatively to 96.19±1.27 at the final follow-up(<0.05). Merle D 'Aubigne score increased from 3.96±0.06 preoperatively to 16.81±0.63 at the final follow-up(< 0.05). No fracture or nerve or vascular injury were found during the operation. The postoperative X-ray showed that the prosthesis was in good position. No complications such as joint dislocation, dislocation of prosthesis, loosening of prosthesis, fracture around the prosthesis, pain in the front of thethigh, fracture of the self tapping screw in the ilium, and delayed infection occurred in the patients after operation.@*CONCLUSION@#Dual mobility total hip prosthesis has the advantages of both good initial stability and low dislocation rate of the prosthesis, and the clinical application of total hip replacement in hemiplegic femoral neck fracture is satisfactory.
Subject(s)
Aged , Female , Humans , Male , Arthroplasty, Replacement, Hip , Femoral Neck Fractures , General Surgery , Follow-Up Studies , Hemiplegia , Hip Prosthesis , Retrospective Studies , Treatment OutcomeABSTRACT
OBJECTIVE@#To investigated the clinical and pathological characteristics of related-renal damage in patients with POEMS syndrome.@*METHODS@#Five patients diagnosed as POEMS syndrome in our hospital were selected. Their clinical manifestation, pathological characteristics of kidney and laboratory examination were analyzed retrospectively. Among the 5 patients, three males and two females with a median age of 50 years old. The mean interval before diagnosis was 13.0±7.2 months.@*RESULTS@#All the patients showed neuropathy, endocrinopathy, monoclonal plasma cell-proliferative disorder, skin changes and extravascular volume overload, in which 4 patients showed organomegaly. Proteinuria was found in 5 patients, and microhematuria was found in 4 patients. Moreover, 4 patients showed an elevated blood urea, while 2 patients showed creatinine elevation. 1 patient at chronic kidney disease (CKD)-G1 stage, 2 patients at CKD-G2 stage, and 1 patient at CKD-G3b stage, moreover, 1 patient at CKD-G5 stage. Endothelial injury and mesangial lesion were the main characteristics of renal pathology. 3 patients were pathologically diagnosed as thrombotic microangiopathy kidney damage, while 2 patients as light chain amyloidosis.@*CONCLUSION@#POEMS syndrome is a multi-systemic disease with complex clinical manifestations. 5 patients had different degrees of renal insufficiency. Endothelial injury and mesangial lesion are the main features of renal pathology.
Subject(s)
Female , Humans , Male , Middle Aged , Kidney , POEMS Syndrome , Paraproteinemias , Renal Insufficiency , Retrospective StudiesABSTRACT
Platelets are non-nuclear blood cells that are widely involved in physiological and pathological processes.Their main role is to participate in hemostasis and thrombosis.Toll-like receptors(TLRs)are innate immune receptors.Platelets express multiple TLRs and can promote thrombosis by recognizing ligand-induced platelet activation and aggregation.This article reviews the relationship between platelets/TLR and thrombosis and the roles of TLRs in the development of thrombotic diseases.