0.05).Conclusion:MBL B allele is not a risk component in the developing process of SLE Chinese patients.

Objective To investigate the application effect of two methods,the nutritional risk screening 2002 (NRS 2002) and the mini nutritional assessment short-form (MNA-SF),on nutritional screening in elderly patients with spontaneous intracerebral hemorrhage,and to evaluate the applicability of two kinds of methods.Methods Using NRS 2002 and MNA-SF,the malnutrition risk screening was performed in 60 cases of elderly patients with cerebral hemorrhage.Based on malnutrition risk score results,the potential nutritional risks were judged,and the application effectiveness was compared between two nutritional evaluation tools.Results Among 60 cases,there were 59 (98.3％) cases with NRS-2002 score≥3,and 1 (1.7％) case with NRS-2002 score ＜ 3.By contrast,using the MNA-SF analysis,among 60 cases there were 3 (5.0％) cases with score of 12-14,7 (11.7％) cases with score of 8-11,and 50 (83.3％) cases with score of 0-7.No statistically significant difference was found between these two methods for analyzing the malnutrition risk incidence (x2 =1.034,P =0.619).Conclusion The risk grade of malnutrition is very high in elderly patients with spontaneous intracerebral hemorrhage during the period of hospitalization.NRS 2002 and MNA-SF are both applicable for the nutritional screening in the elderly patients with spontaneous cerebral hemorrhage.Clinical application is more convenient,fast and comprehensive with MNA-SF than with NRS 2002.

The growing demand for new therapeutic strategies in the medical and pharmaceutic fields has resulted in a pressing need for novel druggable targets. Paradoxically, however, the targets of certain drugs that are already widely used in clinical practice have largely not been annotated. Because the pharmacologic effects of a drug can only be appreciated when its interactions with cellular components are clearly delineated, an integrated deconvolution of drug-target interactions for each drug is necessary. The emerging field of chemical proteomics represents a powerful mass spectrometry (MS)-based affinity chromatography approach for identifying proteome-wide small molecule-protein interactions and mapping these interactions to signaling and metabolic pathways. This technique could comprehensively characterize drug targets, profile the toxicity of known drugs, and identify possible off-target activities. With the use of this technique, candidate drug molecules could be optimized, and predictable side effects might consequently be avoided. Herein, we provide a holistic overview of the major chemical proteomic approaches and highlight recent advances in this area as well as its potential applications in drug discovery.
Chromatography, Affinity ; Drug Delivery Systems ; methods ; Drug Design ; Drug Discovery ; methods ; Humans ; Mass Spectrometry ; Proteome ; chemistry ; Proteomics ; methods ; Small Molecule Libraries ; chemistry

Objective To investigate the effect of flipped classroom model in the theory teaching of urology,and to provide a reference for promoting teaching methods.Methods The clinical medicine classes 1 and 2 of 2014 in the five-year undergraduate program,with 120 students in each class,were enrolled,with class 1 as teaching reform class and class 2 as traditional teaching class.The flipped classroom model was applied in class 1,and the traditional teaching method was applied in class 2.Tests and questionnaires were used at the end of the class to evaluate teaching effect.All the data were analyzed by Graphpad Prism 6.1,t test and Chi-square test were used for stabistical analysis.Results There were significant differences between the teaching reform class and the traditional teaching class in the mean score of chapter test [(80.78± 9.02) vs.(75.03 ± 7.98),P ＜0.05] and the mean score of comprehensive quality [(82.09 ± 8.89) vs.(76.34 ± 6.45),P＜0.05].The questionnaire survey showed an overall degree of satisfaction of 98.5％ among students;81.67％ of all students actively participated in learning,91.67％ were willing to participate in the teaching with flipped classroom model,75.84％ thought this model improved learning enthusiasm,65％ thought it improved the ability of self-study,73.33％ thought it improved their understanding of the learning content,and 84.17％ thought it strengthened the communication between teachers and students.Conclusion The flipped classroom model has a better effect than the traditional teaching model in the theory teaching of urology and is easily accepted by students.Therefore,it holds promise for clinical application.

Objective To establish a human α-synuclein nuclear localization signal transgenic mouse model and investigate the effects of α-synuclein nuclear localization on the behavior of mice.Methods Human α-synuclein nuclear localization signal and EGFP lentiviral vectors were constructed.Transgenic mice were created with the microinjection method.Using PCR and Western Blot method to identify the genotypes and protein expression of the transgenic founder mice and their offsprings.The immunofluorescence was used to examine the localization of human α-synuclein in the mouse brain tissue.The behavioral changes of the transgenic mice were evaluated by the open field test,rotarod test,and O maze test.Results The h SNCA-NLS gene was successfully inserted into the mouse genome,the human α-syn was successfully expressed,and the human α-syn has localized with the nuclear.Further studies found that human α-synuclein nuclear localization signal transgenic mice had significant motor dysfunction,astrocyte proliferation and inflammatory response at 2 months of age and exhibited significant anxiety-like symptoms and reduced expression of the γ-aminobutyric acid(GABA)gene at 9 months of age,which persisted until 12 months of age.Conclusions A human α-synuclein nuclear localization signal transgenic mouse model has been successfully established.The mice exhibit significant motor dysfunction and anxiety-like symptoms.The successful establishment of this model provides a foundation for studying the role of α-syn nuclear localization in Parkinson's disease.

Objective: To observe the effect of differentiated mature adipocytes on hepatic steatosis and aquaporin-9 (AQP9) expressions in HepG2 cells and further explore its possible mechanism of action. Methods: Human preadipocytes were cultured and differentiated to full maturity. HepG2 cells were co-cultured with non-differentiated adipocytes and differentiated mature adipocytes for 48 h, and then labeled as control group and experimental group. Oil red O staining and intracellular triglyceride content were performed on co-cultured HepG2 cells and simultaneous changes in phosphatidylinositol 3-kinase (PI3K) - serine/threonine kinase (Akt) signaling pathway, and AQP9 mRNA and protein levels were detected. The experimental group was co-cultured with recombinant human insulin-like growth factor-I (IGF-I), with the addition of 100ng/ml PI3K-Akt pathway agonist, labeled as experimental group + IGF-I group. The activation of PI3K-Akt pathway was verified by Western blotting (WB). The expression of AQP9 was detected by RT-q PCR and WB. The recombinant lentivirus LV-AQP9 or empty-loaded virus LV-PWPI was transfected with HepG2 cells by recombinant lentiviral transfection tecnique, and labeled as HepG2-AQP9 and HepG2-PWPI. The transfection efficiency was assessed by confocal laser scanning microscopy and RT-qPCR and WB detected the change of AQP9 expression level after virus transfection. Afterwards, the stable over-expressed HepG2-AQP9 cells and the empty-loaded HepG2-PWPI cells were co-cultured with differentiated mature adipocytes for 48h, and labeled as HepG2-AQP9 co-culture group, and then intracellular triglyceride content were detected with Oil red O staining. Finally, IGF-I was added to the HepG2-AQP9 co-culture group, which was recorded as HepG2-AQP9 co-culture + IGF-I group. Intracellular triglyceride content was detected with Oil red O staining, and WB verified PI3K-Akt signaling pathway activation and changes in AQP9 mRNA and protein levels. A t-test was used to compare the two independent samples. Results: The intracellular lipid droplets and triglyceride content (0.052 ± 0.005) in the experimental group was increased significantly than the control group (0.033 ± 0.003) (t= 5.225,P= 0.006), suggesting that adipocyte co-culture had induced steatosis in HepG2 cells. RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (3.615 ± 0.330) and protein levels (0.072 ± 0.005) in the experimental group were significantly higher than the control group (t= 13.708, 11.225,P= 0.005, < 0.001). WB results showed that the expression level of phosphorylated Akt (p-Akt) protein (0.116±0.003) in the experimental group was significantly lower than the control group (0.202 ± 0.003) (t= 27.136,P< 0.001). The total Akt protein was constant, and the p-Akt/total Akt (0.182 ± 0.017)was significantly lower than the control group (0.327 ± 0.019) (t= 2.431,P= 0.001), suggesting that adipocyte co-culture had inhibited PI3K- Akt signaling pathway in HepG2 cells and up-regulated the expression level of AQP9. WB results indicated that the expression level of p-Akt protein (0.194 ± 0.021) in the experimental group + IGF-I group was significantly higher than the experimental group (0.132 ± 0.003) (t= 5.082,P= 0.007). The total Akt protein was constant, and the p-Akt/total Akt (0.281 ± 0.009) was significantly higher than the control group (0.184 ± 0.132) (t= 10.311,P< 0.001). Simultaneously, RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (0.327 ± 0.347) and protein levels (0.042 ± 0.004) in the experimental group + IGF-I group were significantly lower than the experimental group (t= 33.573, 5.598,P< 0.001, 0.005), suggesting that adipocyte co-culture had possibility to regulate the expression level of AQP9 through the PI3K-Akt pathway. Confocal laser microscopy analysis showed that the transfection efficiency was more than 90%. RT-q PCR and WB results indicated that the expression levels of AQP9 mRNA and protein levels (0.373 ± 0.221) in HepG2-AQP9 group were significantly higher than HepG2-PWPI group (t=14.953, 28.931,P= 0.002 and 0.000), suggesting that the stable overexpression of AQP9 cell line was successfully constructed. The intracellular lipid droplets and triglyceride content in HepG2-AQP9 co-culture group was significantly increased (t= 5.478, 5.369,P= 0.005) than HepG2-PWPI co-culture group and HepG2-AQP9 co-culture+ IGF-I group, suggesting that the increased expression of AQP9 had promoted HepG2 steatosis in co-cultured adipocytes. WB results showed the expression levels of p-Akt protein (0.168 ± 0.006) and p-Akt/total Akt (0.265±0.009) in HepG2-AQP9 co-culture + IGF-1 group was significantly increased (t= 16.311, 8.769,P< 0.001) than HepG2-AQP9 co-culture group, while the expression levels of AQP9 mRNA (0.327 ± 0.034) and protein (0.375 ± 0.025) was significantly decreased (t= 33.573, 9.146,P< 0.001 and 0.001). Conclusion Adipocytes co-culture can induce steatosis in HepG2 cells, and may participate in inhibiting PI3K-Akt signaling pathway to upregulate the expression of AQP9 in steatotic HepG2 cells.

Objective: To investigate the application value of real-time virtual sonography(RVS)in the diagnosis and treatment of complicated hepatolithiasis. Methods: The retrospective and descriptive study was conducted. The clinical data of 10 patients with complicated hepatolithiasis who were admitted to Hunan Provincial People′s Hospital between October 2017 and March 2018 were collected. There were 3 males and 7 females, aged from 40 to 69 years, with an average age of 57 years. Patients received abdominal color Doppler ultrasound examination, magnetic resonance cholangiopancreatography, and upper abdominal spiral computed tomography (CT) thinly scanning + enhanced examination. Data of CT examination were imported into RVS. RVS was used to locate hepatolithiasis, relationship between stones and vessels, anatomy of bile ducts and vessels in hepatic hilus. Surgical methods included RVS-guided hilar cholangiotomy, biliary stricturoplasty, bilateral hepatojejunostomy, hepatic segmentectomy (lobectomy), and hepatolithotomy. Observation indicators: (1) surgical and postoperative situations; (2) typical case analysis; (3) follow-up. Follow-up using outpatient examination was performed to detect residual stones up to June 2019. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results: (1) Surgical and postoperative situations: 10 patients underwent RVS-guided surgeries successfully for complicated hepatolithiasis, with successful match in RVS (difference between CT images and ultrosound images <2 mm). No residual stone was identified by choledochoscope during operation. The operation time and volume of intraoperative blood loss were 285 minutes (range, 210-360 minutes) and 200 mL (range, 100-600 mL), respectively. No blood transfusion was needed during the operations. The duration of hospital stay was 20.5 days (range, 14.0-29.0 days). There was no perioperative death. One patient had postoperative biliary leakage and abdominal infection, and was cured after conservative treatment. (2) Typical case analysis: the tenth patient, female, 60 years old, was diagnosed with complicated hepatolithiasis, and was prepared to undergo hepatolithotomy+ quadrate lobectomy and hilar cholangioplasty+ bilateral hepatojejunostomy. Preoperative CT images and intraoperative color Doppler ultrasound images of the patient were fused and matched on the sagittal section of the portal vein and the cross section of the right branch of portal vein, and stones and important vessels were marked on the images. After accurate positioning, a curette was used to remove the stones. Removal of biliary stones through hepatic parenchyma and peripheral dilated bile ducts was conducted at the site where stones obviously existed. After the stones were removed, the intrahepatic bile duct and hilar bile duct merged. The left end of the bile duct split was confirmed by real-time ultrasound. After location of portal vein was determined by ultrasound, vascular plastic surgery was perfomed to avoid stenosis. (3) Follow-up: 10 patients were followed up for 6-12 months, with a median follow-up time of 8 months. One of 10 patients was suspected residual stones at the right peripheral hepatic anterior lobe by postoperative angiography at 2 months after surgery, and was not removed stones by choledochoscope. The patient had no recurrent symptoms after T-tube removal. The other 9 patients had no residual stones. Conclusion RVS applied in complicated hepatolithiasis is helpful for the precise intraoperative diagnosis, and the surgical treatment can be safe and effective.