Objective To assess causal associations between specific gut microbiota and different types of cancer by using the two-sample Mendelian randomization(MR)analysis method.Methods On the basis of summary statistics of gut microbiota from a genome-wide association study(GWAS)conducted in German population(n=8956),single nucleotide polymorphisms(SNPs)that were significantly associated with 430 gut microbiota features were extracted as instrumental variables(Ⅳs).Summary statistics from the GWAS of 17 types of cancer were used as outcomes.Two-sample MR analysis was used to explore the causal relationship between gut microbiota and pan-cancer,where the analysis results were dominated by inverse variance weighting.Meanwhile,sensitivity analyse of heterogeneity and horizontal pleiotropy test were done to keep the stability of results.Results The genetic susceptibility of 17 gut microbiota features was causally associated with the occurrence and development of 11 different types of cancer,respectively.Conclusion By exploring the causal relationship between different gut microbiota features and pan-cancer,this study has found a potential causal relationship between specific gut microbiota features and cancer,and these gut microbiota may become new biomarkers to provide new ideas for cancer prevention,early screening,and treatment.

BACKGROUND:Animal models of diabetic encephalopathy that have been studied mainly include streptozotocin-induced model,high-sugar and high-fat diet-induced model and spontaneous animal model.Establishing a simple,easy,short-cycle,safe and effective model of diabetic encephalopathy can help to explore the subsequent pathogenesis and screen therapeutic drugs. OBJECTIVE:To further explore and evaluate the method of building diabetic encephalopathy rat models. METHODS:Twenty Sprague-Dawley rats were randomly divided into control(n=10)and model(n=10)groups.Rats in the model group were given a single injection of 45 mg/kg streptozotocin in the left lower abdominal cavity,and those in the control group were given the same amount of citrate buffer.During the experiment,the body mass,feed intake,water intake and blood glucose were measured.After 8 weeks,the glucose tolerance and oxidative stress levels were measured,and the pathological changes of brain tissue and the expression of apoptotic proteins were compared between groups. RESULTS AND CONCLUSION:Compared with the control group,the food intake,water intake,encephalization quotient,blood glucose and area under the blood glucose curve were significantly increased in the model group,while the body mass decreased significantly(P<0.01).Histopathological examination of the brain showed that compared with the control group,the number of surviving nerve cells was significantly reduced in the model group(P<0.01),with more significant pathological damage of nerve cells.Compared with the control group,the activities of serum superoxide dismutase,catalase and glutathione in the model group were significantly decreased(P<0.01),and the content of oxidative malondialdehyde was significantly increased(P<0.05).The expression levels of apoptosis-related proteins Bax and Caspase-3 in brain tissue increased in the model group compared with the control group,while the expression of Bcl-2 decreased(P<0.01).In conclusion,an 8-week injection of 45 mg/kg streptozotocin can cause obvious pathological damage to the brain tissue of diabetic rats,to successfully establish the rat model of diabetic encephalopathy.

Objective:To screen human monoclonal antibodies (mAbs) against the nuclear protein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV), identify their binding specificity to both recombinant NP and NP in viral particles, and determine their affinity constant and binding kinetics.Methods:Antibody genes were extracted from the blood of recovered individuals, and an antibody library was created using phage display. This library was panned by recombinant NP. The selected antibodies were expressed and purified. Enzyme linked immunosorbent assay (ELISA), western blot (WB), and indirect immunofluorescence assay (IFA) were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions. Additionally, biolayer interferometry (BLI) was utilized to determine the antibody affinity constant.Results:An antibody library with a capacity of 7.24×10 7 was successfully constructed. Following three rounds of panning, 6 mAbs (named as NP-1, NP-10, NP-11, NP-20, NP-21, and NP-27) were isolated. The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis. Additionally, these mAbs were demonstrated specific in binding to NP in virions as evidenced by IFA detection. The affinity constant values of the 6 mAbs, determined by BLI assay, ranged from 0.47 nmol/L to 32 nmol/L. Conclusions:The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity. These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.

Objective:To screen human monoclonal antibodies (mAbs) against the nuclear protein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV), identify their binding specificity to both recombinant NP and NP in viral particles, and determine their affinity constant and binding kinetics.Methods:Antibody genes were extracted from the blood of recovered individuals, and an antibody library was created using phage display. This library was panned by recombinant NP. The selected antibodies were expressed and purified. Enzyme linked immunosorbent assay (ELISA), western blot (WB), and indirect immunofluorescence assay (IFA) were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions. Additionally, biolayer interferometry (BLI) was utilized to determine the antibody affinity constant.Results:An antibody library with a capacity of 7.24×10 7 was successfully constructed. Following three rounds of panning, 6 mAbs (named as NP-1, NP-10, NP-11, NP-20, NP-21, and NP-27) were isolated. The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis. Additionally, these mAbs were demonstrated specific in binding to NP in virions as evidenced by IFA detection. The affinity constant values of the 6 mAbs, determined by BLI assay, ranged from 0.47 nmol/L to 32 nmol/L. Conclusions:The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity. These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.

Objective:To realize prokaryotic expression, purification and identification of 2019-novel Coronavirus (2019-nCoV) nucleocapsid protein (NP), and apply it to the serological diagnosis.Methods:The synthetic 2019-nCoV NP gene was cloned into the prokaryotic expression vector pET28a to construct expression plasmid, and then purified by Ni-chelating affinity. SDS-polyacrylamide gel electrophoresis (SDS-PAGE), indirect enzyme-linked immunosorbent assay (ELISA), Western blot (WB), and immunochromatography were used to test the purified protein. Indirect ELISA reaction conditions were optimized for serum antibody detection.Results:The relative molecular mass of recombinant NP was about 50×10 3 after SDS-PAGE electrophoresis, which was consistent with the expectation. Indirect ELISA and WB results showed that it could specifically bind to the serum of patients infected with 2019-nCoV. The detection limit of NP was 0.2 ng/ml by immunochromatography. The sera from 32 patients infected with 2019-nCoV and the control sera were detected by indirect ELISA, and the results showed that they were clearly clustered. Conclusions:Prokaryotic expression of 2019-nCoV NP has good immunogenicity and can be used for the development of serological diagnostic reagents.

OBJECTIVE： To establish a method for content determination of total polyphenols from Gastrodia elata， and to optimize the purification technology of macroporous resin. METHODS： The content of total polyphenols from G. elata was determined by Folin-ciocaileu colorimetry. Using the absorption and desorption performance as index， 4 kinds of macroporous adsorption resins were selected by static adsorption and desorption tests. The flow rate and mass concentration of the sampling solution， volume fraction of eluent， eluent flow rate and eluent volume were investigated by dynamic adsorption and desorption tests. The purification technology of macroporous resin was optimized. RESULTS： The linear range of gallic acid was 4-32 μg/mL （r＝0.999 9）. RSDs of precision， stability and repeatability tests were all less than 2％. The recovery rate of the sample was 95.51％-102.94％（RSD＝2.54％，n＝6）. D301 macroporous resin had strong static adsorption and desorption ability from G. elata polyphenols. The optimal purification technology included that the sample solution flow rate 2 BV/h； the sample solution mass concentration 4 mg/mL； the elution solvent 70％ ethanol； the elution flow rate was 3 BV/h， and the eluent volume 5 BV. The content of total polyphenols from G. elata optimized by the optimal purification technology was 0.381 mg/g. CONCLUSIONS： Established method is sensitive and stable. The optimized purification technology is stable and feasible.

Neoantigens, as the products of gene mutations in tumor cells, are specific antigen expressed on the surface of tumor cells. They can be the targets of immuno-cell therapies with high specificity and safety. Immunotherapies based on tumor neoantigens became the new research hotspot as results of the applications of genomic sequencing technologies and the development of neoantigens prediction techniques. In this paper, the applied prospect of neoantigens in hematological malignancies is discussed based on the research progress in the last few years and the relevant reports from the 59th American Society of Hematology Annual Meeting.

Objective To investigate the clinical effect of fluticasone propionate combined with noninvasive positive pressure ventilation in the treatment of chronic obstructive pulmonary disease (COPD).Methods 88 cases of COPD patients from October 2014 to October 2016 in our hospital were selected and randomly divided into two groups of the control group observation group with 44 cases in each group.The control group was treated with conventional treatment, the observation group was treated with fluticasone propionate plus noninvasive positive pressure ventilation, and then the curative effect of the two groups was compared.Results The total effective rate of observation group was 100% higher than that of the control group 70.5%, the difference was statistically significant (P<0.05), after treatment, the observation group of FVC (2.50 +0.32) L, FEV1 (1.36 +0.20) L, FEV1%(51.23+4.32), PaO2(10.51+2.10) kPa, PaCO2(5.15 +1.19) kPa, compared with the control group, the difference was statistically significant (P<0.05), the observation group recurrence rate 4.5% was lower than the control group 13.6%, the difference was statistically significant (P <0.05).Conclusion COPD patients with fluticasone propionate combined noninvasive positive pressure ventilation, can effectively improve lung function, reduce disease recurrence rate, a significant effect.

BACKGROUND:Endplate cartilage degeneration initiates intervertebral disc degeneration. AMP-activated protein kinase (AMPK) regulates the formation and degradation of cartilage. OBJECTIVE:To explore the role of AMPK in an in vitro natural degeneration model of chondrocytes derived from endplate of rat intervertebral discs. METHODS:Morphology of in vitro subcultured endplate chondrocytes of rat intervertebral discs at passages 0, 2, and 5 were observed under an inverted microscope fol owing cytoskeleton staining. Chondrocyte phenotype, proliferation, and the cartilage marker genes (type II col agen, proteoglycan, SOX-9, matrix metal oproteinase-3 and-13), and AMPK phosphorylation were determined by toluidine blue staining, MTT assay, real-time PCR analysis, and western blot assay, respectively. RESULTS AND CONCLUSION:The altered morphology, decreased proliferation ability, and phenotype loss were observed in chondrocytes with increased passage number. Gene expression of type II col agen, proteoglycan, SOX-9 was significantly decreased;while gene expression of matrix metal oproteinase-3 and-13 was significantly increased in endplate chondrocytes at passage 5 compared with those at passages 0 and 2. AMPK phosphorylation in endplate chondrocytes at passage 5 was significantly decreased. These findings indicate that AMPK phosphorylation is involved in in vitro natural degeneration of chondrocytes derived from the endplate of rat intervertebral discs, and the degeneration of endplate chondrocytes and intervertebral discs can be inhibited through the regulation of AMPK activity.

Objective To investigate the safety and efficacy of nephron-sparing surgery (NSS)for selective T2 stage renal tumor.Methods The surgical database of 26 patients treated with NSS for clinical T2 stage renal cell carcinomas between March 2010 and May 2013 were collected and analyzed retrospectively.There were 17 males and 9 females,with a mean age of 52 years (39-74 years),mean tumor size of 10.3 cm(7.2-16.5 cm),and mean R.E.N.A.L score of 7.5 (6-10).Patients'demographics,clinical characteristics,oncologic outcomes,renal function were reviewed.Results The renal masses were removed successfully and the surgical margins were negative.There were 21 (80.8％) cases of clear cell carcinoma,4 (15.4％) papillary carcinoma and 1 (3.8％) chromophobe carcinoma.The mean ischemia time was (28.3 ± 12.5) minutes (7 patients were clamp-free).Three patients needed transfusion,one experienced urine fistula and cured by conservative treatment,and one patient's renal function got progressive worsening and required long-term hemodialysis.The average serum creatinine was 121 μ mol/L before and 136 μmol/L after surgery (P =0.06).After a period of 22-47 months' follow-up,no patient had local recurrence or metastasis.Conclusions NSS can be safely performed and provide effective oncologic outcomes for selective patients with clinical T2 stage renal cell carcinomas.R.E.N.A.L nephrometry is an important factor and should be used to evaluate the feasibility of NSS.