Pachymic acid protects against Crohn's disease-like intestinal barrier injury and colitis in miceby suppressingintestinal epithelial cell apoptosis via inhibiting PI3K/AKT signaling / 南方医科大学学报

OBJECTIVE@#To investigate the effect of pachymic acid (PA) against TNBS-induced Crohn's disease (CD)-like colitis in mice and explore the possible mechanism.@*METHODS@#Twenty-four C57BL/6J mice were randomized equally into control group, TNBS-induced colitis model group and PA treatment group. PA treatment was administered via intraperitoneal injection at the daily dose of 5 mg/kg for 7 days, and the mice in the control and model groups were treated with saline. After the treatments, the mice were euthanized for examination of the disease activity index (DAI) of colitis, body weight changes, colon length, intestinal inflammation, intestinal barrier function and apoptosis of intestinal epithelial cells, and the expressions of TNF-α, IL-6 and IL-1β in the colonic mucosa were detected using ELISA. The possible treatment targets of PA in CD were predicted by network pharmacology. String platform and Cytoscape 3.7.2 software were used to construct the protein-protein interaction (PPI) network. David database was used to analyze the GO function and KEGG pathway; The phosphorylation of PI3K/AKT in the colonic mucosal was detected with Western blotting.@*RESULTS@#PA significantly alleviated colitis in TNBS-treated mice as shown by improvements in the DAI, body weight loss, colon length, and histological inflammation score and lowered levels of TNF-α, IL-6 and IL-1β. PA treatment also significantly improved FITC-dextran permeability, serum I-FABP level and colonic transepithelial electrical resistance, and inhibited apoptosis of the intestinal epithelial cells in TNBS-treated mice. A total of 248 intersection targets were identified between PA and CD, and the core targets included EGFR, HRAS, SRC, MMP9, STAT3, AKT1, CASP3, ALB, HSP90AA1 and HIF1A. GO and KEGG analysis showed that PA negatively regulated apoptosis in close relation with PI3K/AKT signaling. Molecular docking showed that PA had a strong binding ability with AKT1, ALB, EGFR, HSP90AA1, SRC and STAT3. In TNBS-treated mice, PA significantly decreased p-PI3K and p-AKT expressions in the colonic mucosa.@*CONCLUSION@#PA ameliorates TNBS-induced intestinal barrier injury in mice by antagonizing apoptosis of intestinal epithelial cells possibly by inhibiting PI3K/AKT signaling.

Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Carbon nanotube (CNT) composite materials are very attractive for use in neural tissue engineering and biosensor coatings. CNT scaffolds are excellent mimics of extracellular matrix due to their hydrophilicity, viscosity, and biocompatibility. CNTs can also impart conductivity to other insulating materials, improve mechanical stability, guide neuronal cell behavior, and trigger axon regeneration. The performance of chitosan (CS)/polyethylene glycol (PEG) composite scaffolds could be optimized by introducing multi-walled CNTs (MWCNTs). CS/PEG/CNT composite scaffolds with CNT content of 1%, 3%, and 5% (1%=0.01 g/mL) were prepared by freeze-drying. Their physical and chemical properties and biocompatibility were evaluated. Scanning electron microscopy (SEM) showed that the composite scaffolds had a highly connected porous structure. Transmission electron microscope (TEM) and Raman spectroscopy proved that the CNTs were well dispersed in the CS/PEG matrix and combined with the CS/PEG nanofiber bundles. MWCNTs enhanced the elastic modulus of the scaffold. The porosity of the scaffolds ranged from 83% to 96%. They reached a stable water swelling state within 24 h, and swelling decreased with increasing MWCNT concentration. The electrical conductivity and cell adhesion rate of the scaffolds increased with increasing MWCNT content. Immunofluorescence showed that rat pheochromocytoma (PC12) cells grown in the scaffolds had characteristics similar to nerve cells. We measured changes in the expression of nerve cell markers by quantitative real-time polymerase chain reaction (qRT-PCR), and found that PC12 cells cultured in the scaffolds expressed growth-associated protein 43 (GAP43), nerve growth factor receptor (NGFR), and class III β‍-tubulin (TUBB3) proteins. Preliminary research showed that the prepared CS/PEG/CNT scaffold has good biocompatibility and can be further applied to neural tissue engineering research.

Preparation and In Vitro Characterization of Gelatin Methacrylate for Corneal Tissue Engineering

BACKGROUND@#Corneal disease is second only to cataract considered as the leading cause of blindness in the world, with high morbidity. Construction of corneal substitutes In Vitro by tissue engineering technology to achieve corneal regeneration has become a research hotspot in recent years. We conducted in-depth research on the biocompatibility, physicochemical and mechanical properties of rat bone marrow mesenchymal stem cells (rBM-MSCs)-seeded gelatin methacrylate (GelMA) as a bioengineered cornea. @*METHODS@#Four kinds of GelMA with different concentrations (7, 10, 15 and 30%) were prepared, and their physicchemical, optical properties, and biocompatibility with rBM-MSCs were characterized. MTT, live/dead staining, cell morphology, immunofluorescence staining and gene expression of keratocyte markers were performed. @*RESULTS@#7%GelMA hydrogel had higher equilibrium water content and porosity, better optical properties and hydrophilicity. In addition, it is more beneficial to the growth and proliferation of rBM-MSCs. However, the 30%GelMA hydrogel had the best mechanical properties, and could be more conducive to promote the differentiation of rBM-MSCs into keratocyte-like cells. @*CONCLUSION@#As a natural biological scaffold, GelMA hydrogel has good biocompatibility. And it has the ability to promote the differentiation of rBM-MSCs into keratocyte-like cells, which laid a theoretical and experimental foundation for further tissue-engineered corneal stromal transplantation, and provided a new idea for the source of seeded cells in corneal tissue engineering.

Development and Validation of a Prognostic NomogramBased on Clinical and CT Features for Adverse OutcomePrediction in Patients with COVID-19

Objective@#The purpose of our study was to investigate the predictive abilities of clinical and computed tomography (CT)features for outcome prediction in patients with coronavirus disease (COVID-19). @*Materials and Methods@#The clinical and CT data of 238 patients with laboratory-confirmed COVID-19 in our two hospitalswere retrospectively analyzed. One hundred sixty-six patients (103 males; age 43.8 ± 12.3 years) were allocated in thetraining cohort and 72 patients (38 males; age 45.1 ± 15.8 years) from another independent hospital were assigned in thevalidation cohort. The primary composite endpoint was admission to an intensive care unit, use of mechanical ventilation, ordeath. Univariate and multivariate Cox proportional hazard analyses were performed to identify independent predictors. Anomogram was constructed based on the combination of clinical and CT features, and its prognostic performance wasexternally tested in the validation group. The predictive value of the combined model was compared with models built on theclinical and radiological attributes alone. @*Results@#Overall, 35 infected patients (21.1%) in the training cohort and 10 patients (13.9%) in the validation cohortexperienced adverse outcomes. Underlying comorbidity (hazard ratio [HR], 3.35; 95% confidence interval [CI], 1.67–6.71;p < 0.001), lymphocyte count (HR, 0.12; 95% CI, 0.04–0.38; p < 0.001) and crazy-paving sign (HR, 2.15; 95% CI, 1.03–4.48;p = 0.042) were the independent factors. The nomogram displayed a concordance index (C-index) of 0.82 (95% CI, 0.76–0.88),and its prognostic value was confirmed in the validation cohort with a C-index of 0.89 (95% CI, 0.82–0.96). The combinedmodel provided the best performance over the clinical or radiological model (p < 0.050). @*Conclusion@#Underlying comorbidity, lymphocyte count and crazy-paving sign were independent predictors of adverseoutcomes. The prognostic nomogram based on the combination of clinical and CT features could be a useful tool for predictingadverse outcomes of patients with COVID-19.

ATP1 promotes Candida albicans to escape from macrophage killing through regulating oxidative stress / 中华皮肤科杂志

Objective:To investigate the physiological role of F1Fo-ATP synthase α-subunit encoding gene (ATP1) in promoting Candida albicans ( C. albicans) to escape from macrophage killing through eliminating intracellular reactive oxygen species (ROS) by using a reverse genetics approach. Methods:An ATP1 deletion strain and a parental strain of C. albicans were cultured on the YPD media, and the number of formed colonies on the plates was counted to evaluate in vitro viability of C. albicans. To evaluate their in vivo viability, the ATP1 deletion strain and parental strain of C. albicans were inoculated into mice through the caudal vein, kidney tissues were taken out from the mice 1-7 days after the infection, and inoculated onto the YPD medium followed by numeration of colonies after 48 hours of culture. After co-culture of overnight-cultured C. albicans suspensions with macrophages, some of the C. albicans suspensions were inoculated onto the YPD solid medium followed by numeration of colonies and determination of survival rate, and some culture supernatants were inoculated into the 96-well plate for detection of the level of lactate dehydrogenase (LDH) released by macrophages by LDH release assay. A model mimicking oxidative stress in macrophages was established by using hydrogen peroxide. After treatment with hydrogen peroxide, the number of colonies was counted to compare the viability of the C. albicans strains. DCFH-DA staining was conducted to detect the intracellular ROS level in C. albicans after co-culture with macrophages or treatment with hydrogen peroxide, and real-time fluorescence-based quantitative PCR to measure mRNA expression of catalase 1 (CAT1) , superoxide dismutase 4 (SOD4) and SOD5 genes in C. albicans after treatment with hydrogen peroxide. Statistical analysis was carried out by using two-way analysis of variance or Student t test. Results:In vitro, the colony number in both the parental strain group and ATP1 deletion strain group gradually increased over time; after 24 hours, the colony number of the ATP1 deletion strain group was only 10% of that in the parental strain group ( F = 481.84, P < 0.001) . The number of colony formed by the parental strain-infected mouse kidney tissues gradually increased over time, but that by the ATP1 deletion strain-infected mouse kidney tissues gradually decreased, and there was a significant difference between the two groups ( F = 78.27, P = 0.001) . After in vitro co-culture of C. albicans with macrophages, the survival rate in the ATP1 deletion strain group (62.67% ± 3.51%) was significantly lower than that in the parental strain group (82.33% ± 2.52%, t = 7.88, P = 0.001) , and the percentage of LDH released by macrophages was also significantly lower in the ATP1 deletion strain group (27.80% ± 3.54%) than in the parental strain group (87.78% ± 0.17%, t = 33.89, P < 0.001) , which were consistent with the in vivo results. In the model mimicking oxidative stress, the viability of the ATP1 deletion strain group was significantly lower than that of the parental strain group ( F = 3 440.65, P < 0.001) . Both in the co-culture model with macrophages and in the model mimicking oxidative stress in macrophages, the intracellular ROS levels were significantly higher in the ATP1 deletion strain group than in the parental strain group (both P < 0.001) . Furthermore, the mRNA expression of CAT1, SOD4 and SOD5 genes was significantly lower in the ATP1 deletion strain group than in the parental strain group after treatment with hydrogen peroxide (all P < 0.001) . Conclusion:ATP1 deletion may reduce the capabilities of C. albicans to counteract oxidative stress and eliminate ROS, likely by down-regulating the expression of oxidative stress- and ROS clearance-related genes respectively, which may prevent C. albicans from escaping from the macrophage killing and lead it to be eliminated by the host ultimately.

Effect of CDR1/CDR2 or MDR1 genes overexpression on oxidative stress in Candida albicans / 中国人兽共患病学报

To investigate the effect of CDR1/CDR2 or MDR1 genes overexpression on oxidative stress in Candida albicans,we evaluated the effect of H2O2 on cell viability in C.albicans overexpressing genes CDR1 /CDR2 or MDR1 and their parent strains.After establishing an oxidative stress model with H2O2,we detected reactive oxygen species (ROS),mitochondrial membrane potential (Δψm) and the expression of oxidative stress response-related genes (CAP1 and GRP2) and ROS clearance related-genes (SOD2 and SOD5).The results showed that C.albicans growth were inhibited by 100％ after the treatment of 5 mmol/L H2O2.HeO2 caused more ROS accumulation and Δψm reduction in parent strains than in CDR1/CDR2 or MDR1 genes overexpressed strains (P＜0.05).Compared to parent strains,the up-regulated expression of CAP1 and GRP2 were relatively less in CDR1/CDR2 or MDR1 genes overexpressed strains,moreover,the down-regulated expression of SOD2 and SOD5 were also relatively less in CDR1/CDR2 or MDR1 genes overexpressed strains (P＜0.05).In conclusion,the overexpression of CDR1/CDR2 and MDR1 genes could reduce the oxidative stress response and enhance the adaptability of C.albicans to oxidative stress.

The Efficacy and Safety of Endoscopic Esophageal Varix Ligation combined with Omeprazole and Octreotide in the Treatment of Esophageal Variceal Bleeding / 现代生物医学进展

Objective:To explore the efficacy and safety of endoscopic esophageal varix ligation (EVL) combined with omeprazole and octreotide in the treatment of esophageal variceal bleeding.Methods:127 patients with cirrhosis complicated with esophageal variceal hemorrhage diagnosed and treated in our hospital from May 2014 to May 2016 were divided into the study group and the control group.The control group was treated with omeprazole and octreotide on the basis of conventional therapy,while the study group was treated with endoscopic esophageal variceal ligation (EVL) on the basis of control group.The clinical efficacy,hospitalization condition,incidence of adverse reactions and rebleeding rate after treatment of the two groups were analyzed.Results:All the patients in the study group were successfully operated.In the control group,10 patients showed hematemesis and melena,among which 1 patient wastreated with surgery.After treatment,the c lini cal effi cacy of the study group was superi or to the control group,and the di fference was stati sti cally significant (P＜0.05).During the treatment period,the hemostasis time,blood transfusion time,hospital stay and hospitalization expenses of the study gronp were significantly lower than those of the control group (P＜0.05).In the study group and the control group,10 cases and 7 cases respectively had nausea and vomiting,esophageal foreign body sensation,dizziness,palpitations,pain,bloating,increased facces frequency,fever and other adverse reactions,and the incidence of increased faeces frequency of control group was significantly higher than that of the study group (P＜0.05),but the incidence of other adverse reactions and the total incidence showed no significant difference between two groups (P＞0.05).The rate of rebl eeding was si gni fi cantly lower in the study group at 0.5,1,3,6 and 12 months after treatment than those in the control group(P＜0.05).Conclusion:Endoscopic ligation combined with omeprazole and octreotide was effective in the treatment of esophageal variceal bleeding,which could be effective,rapid hemostasis,reduce the hospital stay,hospitalization cost and rebleeding rate with high safety.

In vitro synergistic effect of tetrandrine on ketoconazole against Candida parapsilosis complex / 中华皮肤科杂志

Objective To evaluate the in vitro synergistic effect of tetrandrine on ketoconazole against Candida parapsilosis complex.Methods According to the Clinical and Laboratory Standards Institute (CLSI) M27-A3 guidelines,the microdilution checkerboard method was used to evaluate in vitro antifungal activities of ketoconazole alone and in combination with tetrandrine against 21 clinical isolates of Candida parapsilosis complex based on the fractional inhibitory concentration index (FICI).Antifungal effects of the above drugs at different time points were evaluated by the XTT assay,and then time-killing curves were drawn and assessed to investigate the in vitro dynamic antifungal activity.Results The minimum inhibitory concentrations (MICs) of tetrandrine and ketoconazole alone against 21 clinical isolates of Candida parapsilosis complex were 32-64 mg/L and 0.031 25-2 mg/L,respectively.When ketoconazole was combined with tetrandrine,MICs of tetrandrine and ketoconazole were reduced to 2-8 mg/L and 0.008-0.25 mg/L respectively,and the FICI ranged from 0.09 to 0.5.The time-killing curves revealed that the fungal growth was delayed obviously in the combination group compared with the ketoconazole alone group and tetrandrine alone group.Conclusion Tetrandrine has obvious synergistic effects on ketoconazole against Candida parapsilosis complex in vitro.

Visual quality observation after implantation of TICL for high myopia with astigmatism / 局解手术学杂志

Objective To assess the clinical outcomes about the visual qulitity of toric implantable contact lens ( TICL) implantation for high myopia with astigmatism. Methods Fifty-two eyes of 27 patients that underwent TICL implantation were examined. Uncorrected visual acuity( UCVA) ,best corrected visual acuity( BCVA) ,refraction,contrast sensitivity ( CS) with and without glare were evaluated before and after the treatment. Results Significant improvement in UCVA and BCVA were found at 1 month and 6 months after treatment (P0. 05), and the astigmatism at 1 month ( -0. 35 ± 0. 60)D and 6 months ( -0. 31 ± 0. 42)D after treatment were of no significant difference either (P>0. 05). The CS with and without glare were all significantly better than results before operation for 6. 0,12. 0 and 18. 0 cycles/degree (P0. 05). No significant difference were found preoperatively,1 month after treatment and 6 months after treatment in terms of CS with and without glare (P>0. 05). The satisfaction of this investigation was 100%. Conclusion The TICL performed well in correcting high myopia with astigmatism,and it is a good surgical option for the treatment.

In Vitro Study of Cell Proliferation Inhibition Using Ultrasound Combined with Photodynamic Therapy with Microbubbles Containing Quantum Dots / 中国医学影像学杂志

Purpose To explore the apoptosis mechanism of human ovarian cancer cells SKOV3 processed with ultrasound (US) combined with photodynamic therapy (PDT), with self-contained lactic acid/glycolic acid (PLGA) microbubbles (MBQDs/PLGA) containing quantum dots (QDs) as a photosensitizer. Materials and Methods MBQDs/PLGA was prepared with double emulsion method, and MTT was used to compare the cytotoxic activity difference between QDs and MBQDs/PLGA, to determine the proper treatment condition. SKOV3 cells were treated under selective conditions, cell damage manifestation was observed with HE staining, and double staining flow cytometry was used to detect cell apoptosis. Results When cells were treated with PDT with energy density of 180.0 J/cm2 and US with sound intensity of 0.5 W/cm2 (1.0 MHz, 10 s), cell deformation could be observed under a microscope in both US-PDT-MBQDs/PLGA group and PDT-MBQDs/PLGA group, and cell connections were absent between cells;transmission electron microscope showed dense chromatin gathered along the nuclear membrane, with the formation of apoptotic bodies also be displayed. Maximum apoptosis rate was (22.17±0.38)% Conclusion MBQDs/PLGA-mediated PDT contains proliferation inhibition effect for SKOV3 cells, which can be synergied with low-power ultrasonic irradiation due to sonoporation effect.

Study oil the heredity and variance of E1 gene of rubella virus Matsuba strain / 中华微生物学和免疫学杂志

Objecflve To study the heredity and variance of E1 gene of rubella villls Matsuba vac-cine strain.and to evaluate the protective efficacy of the vaccine prepared by Matsuba vaccine strain on genelevel.Methods The Matsuba strain was passaged on primary rabbit kidnev cells from 14 to 23 generations.and the E1 genes of 14,16,17,18,23 passages amplified by RT-PCR were cloned into the T-A cloning vector pGEM-T and sequenced,respectively.The sequences of E1 gene of the passaged viruses were tom-pared with rubella virus Matsuba reference strain E1 gene(Accession No.D50673)and other rubella strains in GenBank.respectively.Results The Matanba passaged shared higher homology of nucleotide and amino acid with Matsuba reference strain(D50673).,The sequences of nucleotide and amino acid of RV14,RV16.RV18 were identical to D50673.The homology of nucleotide sequences of RV17,RV23 and D50673 was 99.9%and 99.7%,respectively,andthe homology of amino acidwgg 99.8% and 99.2%,respective-ly.The amino acid related to glycosylation and epitopes of the passage viruses were highly conservative.The phylogenetic tree showed Matsuba strain belonged to la genotype.The rubella virus genotypes circulated in China recent years were 1E,lF,2A and 2B,and 1E genotype was the predominant genotype.The homology of nucleotide and amino acid of Matsuba strain and the other genotype reference strains was 92.1%-99.6% and 98.1%-99.8%.respectively.Condusion The Matsuba vaccine strain possesses highly genetic stabil-ity.which indicates that the Matsuba strain and its vaccine are stable on molecular level.And the vaccine prepared by Matsuba vaccine strain can prevent the infection of various genotype rubella viruses.

Evaluation of IRISiQ200 Full Automated Urine Analyzer / 医疗卫生装备

Objective To evaluate the function of IRISiQ200 full automated urine analyzer and its clinical application of user re-classification function.Methods The reproducibility,linearity and mutual infection rate of IRISiQ200 were detected.116 fresh urine specimen were obtained from outpatient and inpatient at random.After the specimen were detected with IRISiQ200,the doubtful urine sediment images were discriminated repeatedly by user reclassification.The microscopic quantitation in uncentrifuged samples was used as reference.Results IRISiQ200 showed good reproducibility,linearity and lower mutual infection rate.The performances by using iQ-200 were: erythrocytes Y=0.596X+11.353,R2=0.834;leukocytes Y=0.468X+5.745,R2=0.708;epithelial cells Y=0.524X-2.649,R2=0.815.The performances after reclassification: erythrocytes Y=0.895X-3.328,R2=0.997;leukocytes Y=0.786X-1.880,R2=0.976;epithelial cells Y=0.911X-5.502,R2=0.992.Conclusion IRISiQ200 full automated urine analyzer shows excellent performance.There is fine correlation between iQ-200 and microscopic quantitation in uncentrifuged samples,and the urine sediment can be observed intuitively by using reclassification function.The doubtful urine sediment images can be discriminated.It can degrade the instrument error and increase the detection accuracy greatly.[Chinese Medical Equipment Journal,2008,29(2):82-83,85]