Objective To analyze the dynamic changes to disease activity,colonic inflammation,histopathology,and serum cytokine levels in mice with chronic ulcerative colitis(UC).Methods For UC induction,2.5％dextran sodium sulfate solution was provided ad libitum for 5 days,and to model remission,tap water was supplied for another 5 days in one induction cycle.Disease activity index(DAI),colon length,and pathological changes to colon tissue were determined.The levels of myeloperoxidase(MPO)in colon tissue and of cytokines such as IL-1 β in serum and colon were detected.Results During the three cycles,disease activity was aggravated and colon length shortened in mice during the induction periods,both of which were relieved during the remission periods.The blood appeared was observed in the stool was earlier in cycles 2 and 3.The number of mice with stool blood increased,and their body weight decreased by a small amount briefly,then recovered rapidly.The degree of histopathological damage to the colon and MPO content in cycles 1 and 3 increased in the induction periods and decreased in the remission periods,with the magnitude of change smaller than that of the change in DAI values;and they increased in the remission period of cycle 2.During induction,the spleen index and serum levels of IL-1β,IL-6,and IL-17A increased continuously and were higher than those in the control group at the end of the experiment.Levels of TNF-α were increased in the induction periods and decreased in the remission periods,and the trend in IL-10 change was similar to that of TNF-α.TGF-β content increased and then decreased and was higher than that in the control group at the end of cycle 3.The colon contents of IL-1β,IL-6,and IL-17A showed similar trends of increasing and then decreasing,but there was no significant change in colon TNF-α.The concentration of IL-10 decreased during the induction periods and increased during the remission periods.Conclusions During the induction of chronic UC in mice,the symptoms of hematochezia and systemic inflammatory reactions gradually increased,and the mice showed an increase in tolerance and ability to resist mortality,weight loss,and histopathological injury to the colon.The onset and remission of colonic histopathological damage lags behind symptomatic changes,and there is a gradual shift in colonic inflammation to a pattern dominated by polymorphonuclear neutrophils(PMN)activation.

Porcine deltacoronavirus(PDCoV),a newly discovered virus in recent years,can cause severe diarrhea,vomiting,dehydration and even death in piglets.S protein is an important structur-al protein of PDCoV,which determines the host or tissue tropism of the virus,and is an important target for the development of PDCoV vaccines and detection methods.In order to prepare mono-clonal antibody(MAb)against the S protein of PDCoV,the recombinant plasmid of S protein was constructed based on the extracellular domain sequence of S protein of PDCoV epidemic strain in China and transformed into DH10Bac competent cells.The recombinant bacmid was identified by blue-white spot screening and PCR.BALB/c mice were immunized with S protein,and the spleen cells of immunized mice were fused with myeloma cells.The positive hybridoma cells that secreted stable antibodies were screened by indirect ELISA and subcloning.Five hybridoma cell superna-tants(MAb)specifically recognizing S protein(2E5,4D5,5D10,2F7 and 5A9)were identified by Western blot and immunofluorescence assay(IFA).Subsequently,the neutralization test showed that three of the monoclonal antibodies(2E5,4D5 and 5D10)could neutralize the virus to varying degrees.The S protein was successfully expressed and 5 monoclonal antibodies that can stably se-crete and specifically bind to PDCoV and S protein were prepared,which laid an important founda-tion for further research on the structure and function of S protein,the development of detection methods for PDCoV infection,and the prevention or treatment of PDCoV infection.

Objective:To investigate the clinical effect of hearing aid combined with cognitive rehabilitation training in the treatment of dementia with hearing loss.Methods:Forty-six cases of dementia patients with hearing loss in Dalian Municipal Central Hospital Affiliated of Dalian Medical University from September 2019 to September 2020 were divided into two groups by random number table method. The treatment group received cognitive rehabilitation training assisted by sound amplifier, while the control group only received cognitive rehabilitation training for 7 weeks. Before and after treatment, questionnaires were used to evaluate the training effect of patients, including MMSE, HAMA, HAMD and ADL, and satisfaction questionnaire was used to evaluate the satisfaction of caregivers.Results:After treatment, the MMSE score of the treatment group was significantly higher than that of the control group: (20.22 ± 3.38) scores vs. (19.89 ± 3.10) scores ( P<0.05), and HAMA and HAMD were significantly lower than those of the control group: (11.43 ± 5.07) scores vs. (11.83 ± 4.79) scores, (8.39±3.37) scores vs. (9.67 ± 5.34) scores ( P<0.05); there was no difference in activities of daily living between the treatment group and the control group ( P>0.05); the caregiver satisfaction of the treatment group was significantly improved, compared with that of the control group ( P<0.05). Conclusions:Hearing assisted cognitive training can improve the cognitive function of patients with hearing-impaired dementia, promote mental and emotional stability, and improve the feelings of caregivers; we emphasize the importance of hearing evaluation for the elderly with dementia, and the importance of cognitive rehabilitation training with hearing assistance.

Objective:To cultivate human-derived prostate cancer (PCa) cells via conditional reprogramming cell (CRC) technology, and establish individualized cell bank for PCa research in vitro.Methods:We obtained three fresh PCa tissue samples from different patients between January 2019 and April 2019. Then each sample was divided into two parts. One was used for cancer nature confirmation by intraoperative biopsy. Another part was sent to the laboratory and digested into single primary cancer cells with 0.25% EDTA enzyme for CRC technology. The details were described as followed: 1. The primary PCa cells were co-cultured with 3T3-J2 cells irradiated by 30 Gy (feeder cells) in conditioned medium, and observed for the growth of cell clones, 2. The feeder cells were removed by 0.25% EDTA trypsin for 1 minute before primary PCa cells digested for passage. All primary PCa cells were validated by multiple experiments such as immunofluorescence, immunohistochemistry, immunoblotting and fluorescence in situ hybridization (FISH).Results:Total three cases of human-derived PCa cell lines were successfully established during 15days through CRC technology. All those primary PCa cells could be steadily and continuously passaged, which also expressed AR, CK5, CK18, P504s and PSA. FISH demonstrated that each cell line harbored≥1.6% TMPRSS2/ERG fusion and conformed to the features of PCa.Conclusion:CRC technology can be used for stable and continuous PCa cell culture in vitro.

Objective To investigate the mechanisms of paeoniflorin in protection of retinal ischemia injury.Methods Fifty-four male specefic pathogen free (SPF) degree Wistar rats were randomly divided into normal control group,model control group and paeoniflorin group.Retinal ischemia injury was induced by raising the intraocular pressure of right eyes of rats to 110 mmHg for 30 minutes.The rats of paeoniflorin group were administrated through intraperitoneal injection of 5 mg/kg paeoniflorin each day for 14 days.OCT and electroretinogram (ERG) were performed to detect the thickness of retinal nerve fiber layer+retinal ganglion cell layer+inner plexiform layer (NGI)and electrophysiological changes of retina,respectively.Retrograde labelling of retinal ganglion cells (RGCs) was used to evaluate the survival number of RGCs.Western blot analysis was used to detect NLRP3,apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC),cleaved caspase 1 (c-caspase 1),IL-18,and IL-1β expression.The use and care of animals complied with the statement of the Association for Research in Vision and Ophthalmology (ARVO) and Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The thickness of retinal NGI in model control group was (58.2 ± 1.7) μm,which was significantly lower than (84.8 ± 1.9) μm in normal control group and (71.1 ±2.4) μm in paeoniflorin group (both at P＜0.05).The amplitudes of A and B waves in paeoniflorin group and normal control group were significantly higher than those in model control group (both at P＜0.05).The number of RGC in model control group was significantly lower than that in paeoniflorin group and normal control group (both at P＜0.05).The relative expressions of NLRP3,ASC,c-caspase 1,IL-18 and IL-1β in model control group were significantly higher than those in normal control group and paeoniflorin group (all at P＜0.05).Conclusions The paeoniflorin can prevent retinal ischemia induced injury of the retina through NLRP3 inflammasomes pathway,which provides a new treatment strategy for clinical therapy.

Objective To retrieve and analyze the guidelines on diabetic foot offloading, and summarize the evidence of high quality guidelines, so as to provide reference for standardizing the offloading treatment of diabetic foot in China. MethodsThe clinical practice guidelines of diabetic foot in the major guideline networks, databases and professional association websites were retrieved by computer. Three researches evaluated the quality of the guidelines, and two researches extracted evidence from documents that met the quality criteria. ResultsA total of 7 guidelines were included, and 5 aspects, 30 recommendations were extracted, including comprehensive treatment, the principle of offloading, offloading of the plantar ulcer, offloading of the non-plantar ulcer, and the gap between the evidence and practice. ConclusionsThis study summarize the best evidence of the diabetic foot offloading therapy, and provides a reference for medical institution to develop appropriate procedures and standardize the practice of diabetic foot offloading therapy. Most of the interventions recommended in the guidelines come from relatively developed countries, but their application may be limited in less developed countries or regions. Therefore, it is urgent to explore offloading methods which have positive offloading effect as well as conform to our national conditions, and apply them to clinical practice gradually in order to improve the therapeutic effect of diabetic foot and improve prognosis.

Objective To investigate the mechanisms of paeoniflorin in protection of retinal ischemia injury. Methods Fifty.four male specefic pathogen free ( SPF) degree Wistar rats were randomly divided into normal control group,model control group and paeoniflorin group. Retinal ischemia injury was induced by raising the intraocular pressure of right eyes of rats to 110 mmHg for 30 minutes. The rats of paeoniflorin group were administrated through intraperitoneal injection of 5 mg/kg paeoniflorin each day for 14 days. OCT and electroretinogram ( ERG ) were performed to detect the thickness of retinal nerve fiber layer+retinal ganglion cell layer+inner plexiform layer ( NGI) and electrophysiological changes of retina, respectively. Retrograde labelling of retinal ganglion cells ( RGCs ) was used to evaluate the survival number of RGCs. Western blot analysis was used to detect NLRP3,apoptosis.associated speck.like protein containing a caspase activation and recruitment domain (ASC),cleaved caspase 1 (c.caspase 1), IL.18,and IL.1β expression. The use and care of animals complied with the statement of the Association for Research in Vision and Ophthalmology ( ARVO ) and Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results The thickness of retinal NGI in model control group was ( 58. 2 ± 1. 7)μm, which was significantly lower than ( 84. 8 ± 1. 9)μm in normal control group and(71. 1±2. 4)μm in paeoniflorin group (both at P<0. 05). The amplitudes of A and B waves in paeoniflorin group and normal control group were significantly higher than those in model control group ( both at P<0. 05 ) . The number of RGC in model control group was significantly lower than that in paeoniflorin group and normal control group ( both at P<0. 05). The relative expressions of NLRP3,ASC,c.caspase 1,IL.18 and IL.1β in model control group were significantly higher than those in normal control group and paeoniflorin group (all at P<0. 05). Conclusions The paeoniflorin can prevent retinal ischemia induced injury of the retina through NLRP3 inflammasomes pathway,which provides a new treatment strategy for clinical therapy.

[Abstract] Objective: To explore the relationship between the preoperative blood indicators (platelets, monocytes, neutrophils-to-lymphocyte ratio) and clinicopathological characters and the prognosis of the early stage malignant melanoma（MM）patients. Methods: Clinicopathological data of 120 cases of stage I-III MM patients, who received initial treatment and radical operation in the Cancer Hospital of Tianjin Medical University from January 2007 to May 2012, were obtained for this study. The correlations between parameters of PLR (platelet-to-lymphocyte ratio), LMR (lymphocyte-to-monocyte ratio), NLR (neutrophil-to-lymphocyte ratio), hemoglobin, neutrophils, lymphocytes, monocytes, eosinophils, basophils, platelets, lactate dehydrogenase, age, stage as well as ulcer and the prognosis of the patients were evaluated. Results: Patients whose tumor with ulceration have higher NLR and basophilic granulocyte (all P< 0.05）. Univariate analysis showed that NLR, PLR, LMR, neutrophil, lymphocyte, monocyte, lactic dehydrogenase, age, stage and ulceration were the risk factors of poor 5-year overall survival (P<0.05). The multivariate analysis identified PLR（HR=4.206， 95%CI:1.65410.696， P<0.01），stage（HR=7.670， 95%CI:3.977-14.795， P<0.01）and ulceration（HR=1.931， 95%CI:1.029-3.623， P<0.05）as independent risk factors for the prognosis of the MM patients. Conclusion: Higher preoperative PLR can be used as a predictive factor for poor prognosis of the early stage MM patients.

OBJECTIVE:To establish a method for the residual simultaneous determination of methanol,alcohol,dichlorometh-ane,n-hexane,tetrahydrofuran and benzene in tosufloxacin tosylat. METHODS:Headspace GC was performed on the capillary col-umn with 6%cyanopropylphenyl-94%dimethylpolysiloxane(DB-624)as fixative lipid,temperature programmed,the inlet temper-ature was 180 ℃,detector was flame ionization detector with temperature of 300 ℃,carrier gas was high purity N2 at a flow rate of 1.5 mL/min,split ratio was 10:1,headspace equilibrium temperature was 100℃,equilibrium time was 40 min,headspace sam-ple volume was 10 mL,and the headspace sample volume was 1 mL. RESULTS:The linear range was 178.3-1782.7 μg/mL for methanol (r=0.9991),301.2-3012.1 μg/mL for alcohol(r=0.9997),33.81-338.10 μg/mL for dichloromethane(r=0.9993), 18.02-180.22 μg/mL for n-hexane(r=0.9991),43.26-432.58 μg/mL for tetrahydrofuran(r=0.9991)and 0.1268-1.2681 μg/mL for benzene(r=0.9991);limits of quantification were 0.31,3.00,0.67,0.02,0.005,0.10 μg/mL,limits of detection were 0.15, 1.51,0.22,0.01,0.001,0.05 μg/mL;RSD of precision test was no higher than 3.1%;RSDs of methanol and n-hexane in stabili-ty and reproducibility tests were no higher than 5%;recoveries were 93.72%-102.20%(RSD=3.1%,n=9),90.10%-101.79%(RSD=4.0%,n=9),97.07%-103.11%(RSD=2.0%,n=9),92.38%-103.83%(RSD=3.9%,n=9),95.44%-103.62%(RSD=2.8%,n=9),and 94.00%-104.73%(RSD=4.1%,n=9). CONCLUSIONS:The method is rapid,sensitive,accurate,and suitable for the residual determination of methanol,alcohol,dichloromethane,n-hexane,tetrahydrofuran and benzene in tosufloxacin tosylat.