Objective To evaluate the effect of nutrition support on nutritional status and clinical outcome of patients at nutritional risk in internal medical departments.Methods 148 patients at nutritional risk as identified by Nutritional Risk Screening 2002 were numbered according to the order of admission and divided into standard care group (control group,odd numbers,n =75) and individualised nutrition support group (intervention group,even numbers,n =73).Intervention consisted of encouraging food intake,designing food plan,and assuring implementation of food prescription.Energy and protein intake,body weight,length of hospital stay,hospitalization expenses and complications were compared between the two groups.Results In the interventions group,protein intake was significantly higher than that in the control group [(45.1 ± 2.2) g/d vs.(54.8±2.5) g/d,P=0.004],and energyintake higher than that in the control group [(4 180.0± 227.4) kJ/d vs.(4 589.6 ± 150.5) kJ/d,P =0.135] but without statistical significance.Intervention led to an intake of ≥75％ of requirements in 46.6％ patients in the intervention group,significantly higher than the proportion in the control group (30.7％) (P =0.047).The change of body weight was significantly smaller in the intervention group than in the control group [(-0.4 ± 0.2) kg vs.(-1.1 ± 0.2) kg,P =0.025].The length of hospital stay,hospitalization expenses,and incidence of complications showed no significant differences between the control group and the intervention group [(13.5 ±0.9) d vs.(12.4 ±0.6) d,P=0.310;(17834±1824) yuanvs.(16099±1243) yuan,P=0.435;12.8％ vs.8.1％,P=0.184].Conclusions Patients at nutritional risk in internal medical departments could benefit from nutrition support in terms of protein intake and body weight maintenance.A large-scale randomized controlled trial is necessary to confirm the effect of nutrition support on clinical outcomes of patients at nutritional risk.

OBJECTIVE: To establish a multidrug-resistant cell line BEL-7402/5-FU of hepatocellular carcinoma (HCC). METHODS: BEL-7402/5-FU was induced by pulse therapy combined with continuous stepwise exposure to 5-fluorouracil in vitro. MTT assay was used to determine its multidrug resistance (MDR). Biological characteristics of the BEL-7402/5-FU cell line were observed including morphological changes, cell growth curve, population doubling time, plate cloning efficiency, adherence rate, cell cycle distribution, chromosome and tumorigenicity. Accumulation amount of adriamycin (ADM) in cytoplasm was measured by flow cytometry. The protein expression of thymidylate synthase (TS) was evaluated by immuno-cytochemical method. RESULTS: The acquired MDR cell line of BEL-7402/5-FU was established successfully. The BEL-7402/5-FU cells showed cross-resistance to ADM, vincristine (VCR), methotrexate (MTX) and oxaliplatin (OHP), whereas still sensitive to hydroxycamptothecin (HCPT). The BEL-7402/5-FU cells tended to grow in clusters in vitro. It was found that the population doubling time of BEL-7402/5-FU cells was longer than that of its parental cells. The plate cloning efficiency and the adherence rate of BEL-7402/5-FU cells at the 2nd and 3rd hour were both lower than those of the parental cells. The distributing proportion of BEL-7402/5-FU cells in G(0)/G(1) phase was less than that of the parental cells, whereas the distributing proportion of BEL-7402/5-FU cells in S phase was higher than that of the parental cells. The accumulation amount of ADM in cytoplasm of BEL-7402/5-FU cells was significantly lower while the expression level of TS protein of which was highly up-regulated as compared with those of the parental cells. CONCLUSION: Establishment of the human HCC cell line BEL-7402/5-FU might be beneficial to the studies of 5-Fluorouracil acquired MDR mechanisms and the selection of reversal modifiers.

OBJECTIVETo investigate the effect of Src kinase inhibitor PP2 on hypoxia/reoxygenation (H/R) injury in rat astrocytes in vitro.

METHODSIn vitro cultured rat astrocytes were exposed to hypoxia for 8 h followed by reoxygenation for 24 h with or without pretreatment with PP2 (10 µmol/L) for 24 h before H/R injury. MTT assay and flow cytometry were used to detect the viability and apoptosis of the exposed astrocytes, respectively, and the protein expressions of Src, Bax, and Bcl-2 in the cells were determined using Western blotting.

RESULTSPP2 pretreatment significantly increased the viability and decreased the apoptosis rate of rat astrocytes exposed to H/R injury (P<0.01). Western blotting showed that H/R injury caused increased expression of Src kinase, which was lowered by PP2 pretreatment. The ratio of Bax/bcl-2 in the astrocytes increased after H/R injury, and was significantly decreased by PP2 pretreatment (P<0.01).

CONCLUSIONPP2 protects rat astrocytes from H/R injury possibly by inhibiting the expression of Src kinase and activating the anti-apoptotic mechanisms in the cells.

Animals ; Apoptosis ; Astrocytes ; pathology ; Cell Hypoxia ; Cells, Cultured ; Flow Cytometry ; Pyrimidines ; pharmacology ; Rats ; src-Family Kinases ; antagonists & inhibitors

The National Medical Licensing Examination has become one of the most important indicator s to measure the teaching quality of medical colleges and universities. In this paper, by analyzing the status of pathophysiology in National Medical Licensing Examination and the current problems existing in pathophysiology teaching, the author proposed a scheme of reform in the pathophysiology teaching based on Medical Licensing Examination, including changing teaching idea, optimizing teaching content, reforming teaching means and adjusting the assessment methods . This reform aims to make the pathophysiology teaching really serve the needs of clinical application.

Objective:To explore the association of platelet to lymphocyte ratio (PLR) and neutrophil to lymphocyte ratio (NLR) with early gastric cancer (EGC), and to assess the predictive value of PLR and NLR in EGC diagnosis.Methods:From January 1, 2017 to December 31, 2020, 178 patients with EGC, 129 patients with chronic gastritis (CG), 122 patients with gastric intraepithelial neoplasia (GIN) admitted and treated at Taizhou Hospital of Zhejiang Province were enrolled. According to Rand random function and with the ratio of 7 to 3, the patients were divided into training group ( n=301, 125 cases of EGC, 90 cases of CG, 86 cases of GIN) and validation group ( n=128, 53 cases of EGC, 39 cases of CG, 36 cases of GIN). The age, gender, routine blood test, carcinoembryonic antigen (CEA) level, Helicobacter pylori ( H. pylori) infection status and other data of the patients were collected. The routine blood test and clinical characteristics of EGC, CG and GIN patients of the training group, and the routine blood test of EGC patients and CG+ GIN patients (hereinafter referred to as non-EGC group) of training group were compared to analyzed the independent risk factors of EGC. Receiver operator characteristic curve (ROC) was drawn. The optimal cut-off value, area under the curve (AUC), OR, 95% confidence interval (95% CI) of independent risk factors were analyzed for EGC diagnosis and prediction. A diagnostic prediction model was established, and the model was apply to the validation group for validation. Hosmer-Lemeshow test was used to test the fitting degree of the model. Compared the AUC of the model applied to training group with validation group to evaluate the discrimination of model. Kruskal-Wallis H test, Mann-Whitney U test or Wilcoxon rank sum test, chi square test, and univariate and multivariate logistic regression analysis were used for statistical analysis. Results:In the training group, the proportions of males and females in CG, GIN and EGC patients were 50.0% (45/90) and 50.0% (45/90), 61.6% (53/86) and 38.4% (33/86), 69.6% (87/125) and 30.4% (38/125), respectively, and the difference was statistically significant ( χ2=8.49, P=0.014). The proportion of males in EGC patients was higher than that in CG patients, and the difference was statistically significant ( χ2 =8.48, P=0.004). The H. pylori infection rate, age, PLR, NLR, lymphocyte count, neutrophil count, and CEA level of CG, GIN and EGC patients in the training group were 18.9% (17/90), 18.6% (16/86) and 43.2% (54/125); 54.0 years old (45.5 years old, 64.0 years old), 63.0 years old (58.0 years old, 66.3 years old) and 66.0 years old (58.5 years old, 71.0 years old); 113.70 (84.48, 136.09), 120.00 (97.94, 138.37) and 124.29 (101.97, 173.57), 1.55 (1.17, 2.23), 1.71 (1.44, 2.02) and 2.04 (1.57, 2.62), 2.00×10 9/L (1.50×10 9/L, 2.40×10 9/L), 1.75×10 9/L (1.50×10 9/L, 2.40×10 9/L) and 1.60×10 9/L (1.30×10 9/L, 2.05×10 9/L), 3.00×10 9/L (2.38×10 9/L, 3.90×10 9/L), 3.00×10 9/L (2.48×10 9/L, 3.40×10 9/L) and 3.30×10 9/L (2.60×10 9/L, 4.30×10 9/L), 1.70 g/L (1.10 g/L, 2.50 g/L), 2.05 g/L (1.48 g/L, 2.90 g/L) and 2.50 g/L (1.55 g/L, 3.40 g/L), respectively, and the differences were statistically significant ( χ2=21.26, H=41.00, 11.79, 21.13, 10.82, 8.54 and 14.42; all P<0.05). The H. pylori infection rate of EGC patients was higher than that of CG and GIN patients, the ages of EGC and GIN patients were older than that of CG patients, the NLR and PLR levels of EGC patients were higher than those of CG patients, the NLR level of EGC patients was higher than that of GIN patients, the level of lymphocyte count of EGC patients was lower than that of CG patients, and the levels of neutrophil count and CEA were higher than those of CG patients, and the differences were statistically significant( χ2=13.98 and 13.90, Z=-6.13, -4.15, -4.07, -3.25, -3.40, -3.18, -2.62 and -3.74; all P<0.017). The levels of PLR, NLR, neutrophil count and CEA of EGC patients were all higher than those of non-EGC patients(124.29 (101.97, 173.57) vs. 117.97 (101.57, 137.32); 2.04(1.57, 2.62) vs.1.66(1.25, 2.17); 3.30×10 9/L (2.60×10 9/L, 4.30×10 9/L) vs.3.00×10 9/L(2.40×10 9/L, 3.60×10 9/L); 2.50 g/L (1.55 g/L, 3.40 g/L) vs. 1.90 g/L(1.23 g/L, 2.70 g/L)), and the lymphocyte count level was lower than that of non-EGC patients (1.60×10 9/L(1.30×10 9/L, 2.05×10 9/L) vs. 1.80×10 9/L(1.50×10 9/L, 2.20×10 9/L)), and the differences were statistically significant ( Z=-3.23, -4.45, -2.91, -3.30 and -2.35; all P<0.05). The results of ROC analysis showed that the optimal cut-off value of PLR, NLR, CEA, neutrophil count and lymphocyte count was 138.18, 1.76, 2.70 g/L, 3.40×10 9/L, 1.80×10 9/L, respectively. The results of univariate analysis indicated that the gender, age, H. pylori infection, neutrophil count, PLR, NLR, lymphocyte count and CEA were all related to EGC ( χ2=5.98, 27.73, 21.26, 8.26, 10.26, 22.80, 4.81 and 25.91; all P<0.05). The results of multivariate analysis demonstrated that age≥70 years old( OR=9.267, 95% CI 3.239 to 26.514), H. pylori infection ( OR=3.353, 95% CI 1.862 to 6.037), NLR >1.76 ( OR=2.084, 95% CI 1.190 to 3.648), PLR>138.18 ( OR=2.452, 95% CI 1.325 to 4.539), CEA >2.70 g/L ( OR=2.637, 95% CI 1.490 to 4.667) were independent risk factors for EGC (all P<0.05). The Hosmer-Lemeshow test showed that there was no statistically significant difference between the predicted value of the model and the actual observed value ( P>0.05), which indicated that the fitting degree of the model was good. In the training group, the AUC of the diagnostic prediction model was 0.787 (95% CI 0.737 to 0.832, P<0.001). The model was applied to the validation group for validation, and the result showed that the AUC of the model was 0.664 (95% CI 0.576 to 0.745, P<0.001), which indicated that the discrimination of the model was good. Conclusions:PLR and NLR are independent risk factors of EGC, and may help to identify EGC. In this study the established diagnostic model has good discrimination and fitting degree, which can provide important reference information for early clinical diagnosis of EGC, which may facilitate early treatment and improve prognosis of patients.

Objective To evaluate the correlation between cerebral blood flow perfusion and memory impairment in patients with severe stenosis of vertebral basilar artery (VBA).Methods 62 cases of patients with VBA stenosis diagnosed by digital subtraction angiography(DSA) in Beijing Tiantan Hospital from September 2016 to March 2017 were enrolled.Mental State Examination (MMSE),Clinical Memory Scale (CMS) test and CT perfusion(CTP) was performed.All patients were divided into memory normal group(n=24,including 1 excellent case,6 above normal cases,and 14 normal cases) and memory impairment group(n =38,including 18 below normal cases,12 periphery cases,8 impaired cases) according to CMS.The ratios of side-to-side period were compared between bilateral mesial temporal lobe and anterior circulation area.The relative time to peak (rTTP),relative mean transit time(rMTY),relative cerebral blood flow(rCBF) and relative cerebral blood volume (rCBV) were calculate.Results The incidence of CTP decompensation in the medial temporal lobe was higher than that in the patients with memory impairment(P＜0.05).The difference of rTTP and rMTT value between the two groups in the bilateral medial temporal lobes was statistically significant (rTFP:(1.131 ±0.037),(1.437±0.139),t=10.520,P＜ 0.05);rMTT:(1.081 ±0.059),(1.281 ±0.174),t=5.423,P＜0.05).Conclusion The patients with VBA severe stenosis are more likely to get memory impairment due to cerebral hypoperfusion.

ObjectiveTo observe the effect of icariin on the recombinant Ras homolog family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway in rats with Alzheimer's disease （AD）， and to explore the mechanism of icariin in ameliorating the neuronal and dendritic damage. MethodThe β-amyloid 1-42 （Aβ_1-42， 2.5 g·L^-1） was used to induce AD in rats via lateral ventricle injection， and the rats were divided into a model group， a low-dose icariin group （0.03 g·kg^-1）， a middle-dose icariin group （0.06 g·kg^-1）， a high-dose icariin group （0.09 g·kg^-1）， and a control group. The control group and the model group were given an equal volume of normal saline at a dose of 10 mL·kg^-1. The cognitive function of rats was assessed by the Morris water maze. The pathological morphology of the rat hippocampal CA1 area was observed by Nissl staining. Dendritic spine density and dendritic length in the CA1 region of the hippocampus were observed by Golgi-Cox staining. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， IL-6， RhoA， ROCK1， and ROCK2 in the hippocampus. Western blot assay was used to detect the protein expression levels of TNF-α， IL-1β， IL-6， RhoA， ROCK1， and ROCK2 in the hippocampus. ResultAs compared with the control group， the escape latency of the rats in the model group was increased （P<0.01）， while the number of crossing the platform and the dwelling time in the target quadrant were decreased （P<0.01）. As compared with the model group， the escape latency of the rats in the middle and high-dose icariin groups was decreased （P<0.05， P<0.01）， while the number of crossing the platform and the dwelling time in the target quadrant were increased （P<0.05， P<0.01）. As compared with the control group， the number of neurons， dendritic spine density， and dendritic length in the hippocampal CA1 area of the rats in the model group were decreased （P<0.01）. As compared with the model group， the number of neurons， dendritic spine density， and dendritic length in the hippocampus of the rats in the middle and high-dose icariin groups were increased （P<0.05， P<0.01）. As compared with the control group， the mRNA and protein expression levels of TNF-α， IL-1β， IL-6， RhoA， ROCK1， and ROCK2 in the hippocampus of the rats in the model group were increased （P<0.01）. As compared with the model group， the mRNA and protein expression levels of TNF-α， IL-1β， IL-6， RhoA， ROCK1， and ROCK2 in the hippocampus of the rats in the middle and high-dose icariin groups were decreased （P<0.05， P<0.01）. ConclusionIcariin improves cognitive function and neuronal and dendritic damage in AD by inhibiting the RhoA/ROCK signaling pathway.

Tumor-specific neoantigens have attracted much attention since they can be used as biomarkers to predict therapeutic effects of immune checkpoint blockade therapy and as potential targets for cancer immunotherapy. In this study, we developed a comprehensive tumor-specific neoantigen database (TSNAdb v1.0), based on pan-cancer immunogenomic analyses of somatic mutation data and human leukocyte antigen (HLA) allele information for 16 tumor types with 7748 tumor samples from The Cancer Genome Atlas (TCGA) and The Cancer Immunome Atlas (TCIA). We predicted binding affinities between mutant/wild-type peptides and HLA class I molecules by NetMHCpan v2.8/v4.0, and presented detailed information of 3,707,562/1,146,961 potential neoantigens generated by somatic mutations of all tumor samples. Moreover, we employed recurrent mutations in combination with highly frequent HLA alleles to predict potential shared neoantigens across tumor patients, which would facilitate the discovery of putative targets for neoantigen-based cancer immunotherapy. TSNAdb is freely available at http://biopharm.zju.edu.cn/tsnadb.
Antigens, Neoplasm ; metabolism ; Data Analysis ; Databases, Genetic ; Humans ; Immunotherapy ; Mutation ; genetics ; Neoplasms ; genetics ; immunology ; Tumor Suppressor Protein p53 ; genetics ; Urinary Bladder Neoplasms ; genetics

BRAF is a serine/threonine kinase that harbors activating mutations in ∼7% of human malignancies and ∼60% of melanomas. Despite initial clinical responses to BRAF inhibitors, patients frequently develop drug resistance. To identify candidate therapeutic targets for BRAF inhibitor resistant melanoma, we conduct CRISPR screens in melanoma cells harboring an activating BRAF mutation that had also acquired resistance to BRAF inhibitors. To investigate the mechanisms and pathways enabling resistance to BRAF inhibitors in melanomas, we integrate expression, ATAC-seq, and CRISPR screen data. We identify the JUN family transcription factors and the ETS family transcription factor ETV5 as key regulators of CDK6, which together enable resistance to BRAF inhibitors in melanoma cells. Our findings reveal genes contributing to resistance to a selective BRAF inhibitor PLX4720, providing new insights into gene regulation in BRAF inhibitor resistant melanoma cells.