Objective:To explore the relevancy between the uridine diphosphate-glucuronylgly-cosyltransferase 1A1 (UGT1A1) gene mutation and the phenotype of indirect hyperbilirubinemia in children.Methods:Sixteen cases with indirect hyperbilirubinemia who visited the Department of Gastroenterology, Children's Hospital of Nanjing Medical University from July 2013 to November 2019 were retrospectively analyzed and were divided into Gilbert syndrome (GS), Crigler-Najjar syndrome type II (CNS-II), and indirect hyperbilirubinemia groups unexplained by UGT1A1 gene mutations. The differences in gene mutation site information and general clinical data were compared. The association between gene mutation spectrum and bilirubin level was explored by t-test analysis.Results:Ten of the sixteen cases with indirect hyperbilirubinemia had GS, three had CNS-II, and three had indirect hyperbilirubinemia unexplained by UGT1A1 gene mutations. A total of six mutation types were detected, of which c.211G?>?A accounted for 37.5% (6/16), c.1456T?>?G accounted for 62.5% (10/16), and TATA accounted for 37.5% (6/16), respectively. Compared with the GS group, the CNS group had early disease onset incidence, high serum total bilirubin ( t ?=?5.539, P ??0.05) and age of onset ( t ?=?0.3611, P ?>?0.05) between the two groups. There was no significant correlation between the number of UGT1A1 gene mutations and serum bilirubin levels. Children with c.1456T?>?G homozygous mutations had the highest serum bilirubin levels. Conclusion:The common pathogenic variants of the UGT1A1 gene sequence are c.1456T?>?G, c.211G?>?A, and TATA, indicating that these site mutations are related to the occurrence of indirect hyperbilirubinemia and have important guiding significance for the etiological analysis of indirect hyperbilirubinemia in children.

Objective:To investigate the clinical efficacy of neuroendoscopic hematoma removal versus soft channel drainage in the treatment of chronic subdural hematoma. Methods:The clinical data of 102 patients with chronic subdural hematoma who received treatment in Jincheng People's Hospital from May 2018 to May 2020 were retrospectively analyzed. They were divided into the neuroendoscopy group ( n = 50) and the soft channel group ( n = 52) according to different surgical methods. Perioperative indexes, hematoma clearance rate, China Stroke Scale score, the activity of daily living score, and oxidative stress indexes were compared between the two groups. All patients were followed up for 3 months. The incidence of complications during the follow-up period was calculated. Results:The retention time of the drainage tube in the neuroendoscopy group was shorter than that in the soft channel group [(2.45 ± 0.63) days vs. (3.30 ± 0.78) days, t = 6.06, P < 0.001]. The length of hospital stay in the neuroendoscopy group was shorter than that in the soft channel group [(7.14 ± 1.65) days vs. (9.07 ± 2.11) days, t = 5.15, P < 0.001]. The hematoma clearance rate at postoperative 7 days in the neuroendoscopy group was higher than that in the soft channel group [(93.45 ± 5.50)% vs. (81.86 ± 7.24)%, χ2 = 9.12, P < 0.001]. There were no significant differences in operation time and intraoperative blood loss between the two groups (both P > 0.05). At postoperative 30 days, the China Stroke Scale score in the neuroendoscopy group was lower than that in the soft channel group [(12.74 ± 2.23) points vs. (18.67 ± 2.45) points, t = 12.79, P < 0.001]. The activity of daily life score in the neuroendoscopy group was significantly higher than that in the soft channel group [(77.69 ± 7.11) points vs. (91.35 ± 7.25) points, t = 9.60, P < 0.001]. At postoperative 7 days, glutathione peroxidase level in the neuroendoscopy group was significantly lower than that in the soft channel group [(130.75 ± 13.66) U/L vs. (148.60 ± 14.64) U/L, t = 6.37, P < 0.001]. Malondialdehyde level in the neuroendoscopy group was significantly lower than that in the soft channel group [(5.11 ± 0.65) nmol/L vs. (6.19 ± 0.74) nmol/L, t = 7.83, P < 0.001]. Superoxide dismutase level in the neuroendoscopy group was significantly higher than that in the soft channel group [(275.60 ± 22.33) U/L vs. (254.60 ± 18.55) U/L, t = 5.15, P < 0.001]. There was no significant difference in the incidence of complications between the two groups ( P > 0.05). Conclusion:Compared with soft channel drainage, neuroendoscopic hematoma removal can obtain better short-term curative effects and less oxidative stress response in the treatment of chronic subdural hematoma. Neuroendoscopic hematoma removal does not increase the incidence of postoperative complications and is highly safe.

OBJECTIVE To provide reference for the regulation of electronic prescription circulation under the mode of “medical science + pharmacy”. METHODS Based on evolutionary game theory， a tripartite evolutionary game model was constructed for related parties of electronic prescription circulation （government agencies， internet hospitals and third-party drug platforms） under the mode of “medical science + pharmacy”. The influential factors of the three parties’ strategy selection were analyzed and verified by simulation. RESULTS & CONCLUSIONS The strategic choices of all parties in the game were affected by the other two parties. Reducing the cost of strict supervision or increasing the punishment for the other two parties would increase the probability of strict supervision of government agencies. Enhancing reputation losses， increasing penalties， and reducing the cost difference when choosing different strategies would have regulatory and binding effects on internet hospitals and third-party drug platforms. It is suggested that government agencies can introduce diversified supervision mode and establish an integrated “intelligent supervision + traceability” system； pay attention to the construction of reputation mechanism， and promote the coordinated linkage of all parties； increase the punishment for violations， provide policy support， reduce the cost of compliance behavior to promote the high-quality development of electronic prescription circulation under the mode of “medical science + pharmacy”.

Objective: To evaluate the value of MRI-T₂WI texture analysis in the differentiation of atypical medulloblastoma and ependymoma of the fourth ventricle. Methods: Preoperative MRI data of 36 cases of fourth ventricle tumor (19 cases of medulloblastoma and 17 cases of ependymoma) confirmed by the Central Theater General Hospital of the Chinese People′s Liberation Army were retrospectively analyzed. Manually sketch areas of interest (ROI) were made using texture analysis software to get histogram parameters, including mean, median, standard deviation, skewness, kurtosis, maximum, minimum, heterogeneity, entropy, the 5th percentile (T₂WI_5th), the 10th percentile (T₂WI_10th), the 25th percentile (T₂WI_25th), the 50th percentile (T₂WI_50th), the 75th percentile(T₂WI_75th), the 95th percentile(T₂WI_95th). Independent sample t test or mann-whitney U test was used for statistical analysis of each parameter value between the two groups. Logistic regression analysis was used to perform multiparameter joint analysis on meaningful parameters. The area under the curve (AUC) was obtained by using the subject operating characteristic (ROC) curve to determine the threshold and diagnostic efficacy for distinguishing the two group. Results: The mean, median, T₂WI_5th, T₂WI_10th, T₂WI_25th, T₂WI_50th, T₂WI₇5th values of the medulloblastoma group were smaller than those of the ependymoma group, and difference was statistically significant (P<0.05). The T₂WI_5th had the best diagnostic performance. When the critical value was 1 343, the diagnostic specificity was 75.0% and the sensitivity was 92.9%. Logistic regression was used to predict the joint analysis of probability parameters. T₂WI_5th+T₂WI_10th had the best performance, and AUC was 0.928. When the critical value was 0.75, the diagnostic specificity was 75.0% and the sensitivity was 100.0%. Conclusions MRI-T₂WI texture analysis can provide quantitative information for preoperative diagnosis of atypical medulloblastoma and ependymoma and improve the accuracy of preoperative diagnosis, which has obvious clinical value.

BACKGROUND:Fibroblast-like synoviocytes (FLS) in the synovial lining layer are related to the cell proliferation, invasion, migration and apoptosis as well as bone resorption in rheumatoid arthritis. OBJECTIVE:To compare the migration and invasion abilities of FLS (MH7A) in rheumatoid arthritis and normal FLS (HFLS). METHODS:The capacities of cell migration and invasion were evaluated by Transwell cell migration and invasion assays. The primers of the indicated microRNAs were designed and synthesized, and the expression levels of miRNAs were determined by real-time PCR according to the SYBR?PrimeScript?miRNA RT-PCR Kit instruction. RESULTS AND CONCLUSION:MH7A possessed stronger migration and invasion abilities than HFLS. Compared with HFLS, obviously upregulated miR-132, -155, -203, -223 and -124, and significantly downregulated miR-15a, -16, 18a, -19a, -26a and -146a were found in MH7A. These findings suggest that the differentially expressed 11 kinds of rheumatoid arthritis-associated miRNAs participate in the pathogenesis of rheumatoid arthritis probably by enhancing the migration and invasion capacities of MH7A.

OBJECTIVE To investigate the effect and mechanisms of chrysophanol on the expression of c-fos and c-jun mRNA induced by ammonia chloride(NH4Cl) in mouse astrocytes. METHODS Primary mouse astrocytes were cultured with NH4Cl 5 mmol·L-1+chrysophanol 0.1,1.0 or 10.0 mg·L-1 ,or NH4Cl 5 mmol · L- 1+extracellular regulated kinase1/2(ERK1/2) inhibitor UO126 10 mmol · L- 1 and NH4Cl 5 mmol · L-1+p38 mitogen-activated protein kinase(p38 MAPK)inhibitor SB239063 10 mmol · L-1 for 48 h. The levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),malondaldehyde(MDA), nitric oxide(NO)and nitric oxide synthase(NOS)were detected by UV spectrophotometry. The phos?phorylation levels of ERK1/2 and p38 MAPK were detected by Western blotting. The expression of c-fos and c-jun mRNA was detected by RT-PCR. RESULTS Compared with NH4Cl 5 mmol · L- 1 group, ammonia-induced astrocytes oxidative stress was improved by chrysophanol (1.0 and 10.0 mg · L-1). The content of MDA and NO and the activity of NOS were reduced(P<0.05). The activity GSH-Px and SOD was increased(P<0.05). The phosphorylation level of ERK1/2 and p38 MAPK induced by ammonia was reduced in chrysophanol groups (P<0.05). Ammonia-induced c-fos and c-jun mRNA expression downregulation was reversed by chrysophanol (0.1,1.0 and 10.0 mg · L-1)and UO126 and SB239063(P<0.05). CONCLUSION Chrysophanol may improve the downregulated expresion of c-fos and c-jun mRNA in mouse astrocytes exposed to NH4Cl by anti-oxidative stress by inhibiting the ERK1/2 and p38 MAPK phosphorylation.

Objective:To compare drug policies in Brazil, Russia, India, China, and South Africa (BRICs), sum-marize the experiences from the process of implementation and provide some suggestions to further improve the national drug policy in China. Methods:National drug polices of BRICs and experiences in the implementation of essential medicine policy were summed up systematically by means of systematic review. Results:Essential medicine policy and popular phar-macy are implemented in Brazil;Pharmaceutical separation system is established in Russia;the mode of essential medicine policy in Delhi is typical in India;Government of South African focuses on procurement and distribution of drugs. Although drug policies of BRICs have their own characteristics, the objective is to increase the public access to low-cost, high-quality drugs. Conclusions:Despite the BRICs have attained some achievements, they still face serious challenges in deep-ening the health care reform. Hence, they need to explore the measures continuingly consisting with their national condi-tions, and learn from the experiences of developed countries to further consolidate and improve the national drug policies.

Objective To compare the diagnostic value of CT and diffusion-weighted imaging in extremity soft tissue tumors. Methods A total of 104 cases of extremity soft tissue tumors were examined with CT scanning and MRI. All cases were histologically proven. Then we compared the CT value of various types of tumors. The b values of diffusion were 0 and 500 s/mm2. The apparent diffusion coefficient (ADC) values of a large region with no hemorrhage, necrosis, scar tissue, or calcification representing the lesion were measured. ADC values of benign tumors, malignant tumors and normal muscles were compared. Results There were 68 cases of benign tumors and 36 cases of malignant tumors. The CT findings of 45 cases and the MRI findings of 87cases were in accordance with pathological examination. The diagnosis of 59 cases by CT and 17 cases by MRI were wrong. The CT features of soft tissue tumors showed the low density masses. The features of lipoma or cyst were typical on CT. There were large differences among the different types of tumors performance on T1WI and T2WI. The ADC values of the malignant tumors were significantly lower than those of benign lesion sand muscles (P < 0.01). There was no significant difference in ADC values between benign lesions and muscles. there was significant difference between the detection level of CT and MRI (P < 0.01). Conclusion CT can clearly show soft tissue tumor lesions and to clarify their relationship and the surrounding tissue, but can not accurately characterize. MRI diffusion-weighted imaging can better differentiate benign and malignant, and speculate the histological lesions sources. MRI detection level is significantly higher than CT and more consistent with a higher degree of pathology. Thus in the preoperative diagnosis of soft tissue tumors, diffusion-weighted imaging MRI should be preferred.

OBJECTIVETo study on the separation from essential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride by ultrafiltration and acetoacetate extraction methods respectively, and the comparison of the oil yields and chemical compositions.

METHODEssential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride was separated by ultrafiltration and acetoacetate extraction methods respectively, and the chemical compositions were analyzed and compared by GC-MS.

RESULTUltrafiltration method could enrich essential oil more and its chemical compositions were more similar to the essential oil prepared by steam distillation method.

CONCLUSIONUltrafiltration method is a good medium to separate essential oil from essential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride.

Chemical Fractionation ; methods ; Citrus ; chemistry ; Emulsions ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Ultrafiltration ; methods

BACKGROUND: A great quantity of cell loss in early stage following stem cell transplantation can significantly affect transplantation effect. Presently, it is confirmed that overexpression of AKT1 gene significantly inhibit cell apoptosis. OBJECTIVE: To explore whether AKT1 gene overexpression can block stem cell apoptosis under hypoxic condition following pig autologous bone marrow mesenchymal stem cell (BMSC) transplantation, and the effect of repairing damaged myocardium. DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Soochow University from August 2005 to February 2007.MATERIALS: A total of 24 healthy male Meishan pigs were supplied by the Animal Experimental Center of Soochow University. METHODS: The CDS (regulation domin of AKT1) AKT1-cDNA fragment was amplified. Lentivector Packaging Kit was used to transfect BMSCs after synthesized with pCDH1-AKT1 shuttling plasmid. Following BrdU labeling, models of myocardial infarction were constructed by occluding the distal left anterior descending coronary artery in pigs with gelatin sponge. 4 weeks later, pigs were randomly divided into four groups: the model control group, the DMEM group, the BMSCs group, and the AKT-transfected group. In model control group, there was no other injection after occluding the left anterior descending coronary artery. In the DMEM group, 5 mL DMEM was injected into the coronary artery. 5 mL BMSCs (1×10~7 cells) were infused into the coronary artery in the BMSCs group. 5 mL BMSCs transfected with the AKT1 gene were injected in the AKT-transfected group MAIN OUTCOME MEASURES: Western blot analysis and real time RT-PCR were used to test the plasmid. The cardiac function was evaluated by magnetic resonance image. Histological characteristics of the myocardium were observed using immunohistochemistry. Serum vascular endothelial growth factor and transforming growth factor β1 levels were determined by ELISA. RESULTS: AKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed in comparison with the published one. AKT mRNA expression could be detected distinctly 24 and 48 hours after transfecting cells. The expression of AKT1 intensity in MSCs remained strong 2 weeks later with detected by real time RT-PCR and Western blot analysis. AKT1-mRNA transcriptional levels were 120 times of primary cells. Before the cell implantation, the left ventricular end-diastolic dimension increased and the stroke volume decreased in the myocardial infarction hearts. The cardiac function was significantly improved after cell implantation, and the implanted MSCs prevented the infarct region from thinning and expanding, improved contraction and increased perfusion in all groups relative to the control hearts. The left ventricular chamber size was smaller in the hearts with being transplanted cells than that in the control hearts. Moreover, the improvement was even markedly greater in AKT-transfected group (P < 0.05). Hematoxylin-eosin staining results showed that fibering was significant in the model control group and DMEM group. Island-like myocardium was observed in the infarct zone of the BMSCs group and AKT-transfected group, and plenty of small vessels-shape structure was detected in the AKT-transfected group. Immunohistochemistry demonstrated that Von Willebrand Factor (vWF) and Cx-43 expression was determined in the myocardium in the BMSCs group and AKT-transfected group, and the proportion of BrdU and Cx-43-positive cells to BrdU-positive cells was significantly greater in the AKT-transfected group compared with the BMSCs group 4 weeks following transplantation (P < 0.05). Following cell transplantation, vascular endothelial growth factor levels were gradually increased, peaked at 1 week, gradually decreased, and reached a normal level at 4 weeks. Transforming growth factor p1 levels were gradually reduced, and significantly less than the model control group, DMEM group 4 weeks later (P < 0.05), and significantly lower than that pretransplantation (P < 0.05).CONCLUSION: Using lentiviral vector to construct with AKT1 gene could stably make BMSCs overexpress AKT1. The BMSCs engraftment in host myocardium might improve the left ventricle function by attenuating the contractile dysfunction and pathologic thinning in this model of left ventricular wall infarction. AKT1 overexpression can significantly improve cardiac function following infarction.