OBJECTIVES: To explore the mechanism of Lacticaseibacillus paracasei E6 for improving vinorelbine-induced immunosuppression in zebrafish. METHODS: The intestinal colonization of L. paracasei E6 labeled by fluorescein isothiocyanate (FITC) in zebrafish was observed under fluorescence microscope. In a zebrafish model of vinorelbine-induced immunosuppression, the immunomodulatory activity of L. paracasei E6 was assessed by analyzing macrophage and neutrophil counts in the caudal hematopoietic tissue (CHT), the number of T-lymphocyte, and the expressions of interleukin-12 (IL-12) and interferon-γ (IFN-γ). The contents of short-chain fatty acids (SCFAs) in L. paracasei E6 fermentation supernatant and the metabolites of L. paracasei E6 in zebrafish were detected by LC-MS/MS-based targeted metabolomics. The immunomodulatory effects of the SCFAs including sodium acetate, sodium propionate and sodium butyrate were evaluated in the zebrafish model of immunosuppression. RESULTS: After inoculation, green fluorescence of FITC-labeled L. paracasei E6 was clearly observed in the intestinal ball, midgut and posterior gut regions of zebrafish. In the immunocompromised zebrafish model, L. paracasei E6 significantly alleviated the reduction of macrophage and neutrophil counts in the CHT, increased the fluorescence intensity of T-lymphocytes, and promoted the expressions of IL-12 and IFN-γ. Compared with MRS medium, L. paracasei E6 fermentation supernatant showed significantly higher levels of acetic acid, propionic acid and butyric acid, which were also detected in immunocompromised zebrafish following treatment with L. paracasei E6. Treatment of the zebrafish model with sodium acetate and sodium propionate significantly increased macrophage and neutrophil counts in the CHT and effectively inhibited vinorelbine-induced reduction of thymus T cells. CONCLUSIONS L. paracasei E6 can improve vinorelbine-induced immunosuppression in zebrafish through its SCFA metabolites acetic acid and propionic acid.
Animals ; Zebrafish/immunology* ; Acetic Acid/metabolism* ; Propionates/metabolism* ; Fatty Acids, Volatile/metabolism*

OBJECTIVES: To investigate the efficacy of Lactobacillus plantarum ZG03 (L. plantarum ZG03) for ameliorating oxidative stress in zebrafish. METHODS: We evaluated the growth pattern of L. plantarum ZG03, observed its morphology using field emission scanning electron microscopy, and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis. FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish. In a zebrafish model of 2% glucose-induced oxidative stress, the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue (CHT), superoxide dismutase (SOD) activity, reactive oxygen species (ROS) levels, and malondialdehyde (MDA) content. Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids (SCFAs) in the zebrafish, and the antioxidant effects of the key metabolites (acetate, propionate, and caproate) were tested. RESULTS: On MRS agar, L. plantarum ZG03 formed circular, smooth, moist, and milky-white colonies with a rod-shaped cell morphology. Genomic analysis revealed abundant sugar metabolism gene clusters. After inoculation of FITC-labeled L. plantarum ZG03 in zebrafish, green fluorescence was clearly observed in the intestinal bulb, mid-intestine, and hind intestine. In zebrafish with glucose-induced oxidative stress, L. plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity. L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid, propionic acid, and caproic acid in zebrafish metabolites. In addition, sodium acetate, sodium propionate, and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models. CONCLUSIONS L. plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites, particularly the SCFAs including acetic acid, propionic acid and caproic acid.
Animals ; Zebrafish/metabolism* ; Oxidative Stress ; Lactobacillus plantarum/metabolism* ; Fatty Acids, Volatile/metabolism* ; Probiotics ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/metabolism*

Objective:To preliminarily assess the clinical value of sirolimus combined with prednisone in the treatment of Kaposiform hemangioendothelioma (KHE) complicated by Kasabach-Merritt phenomenon (KMP) .Methods:A retrospective study was conducted. General clinical data and relevant data on efficacy and adverse reactions were collected from 36 patients with KHE complicated by KMP, who received oral sirolimus combined with prednisone in Department of Pediatric Surgery, West China Hospital, Sichuan University from January 2011 to January 2018.Results:Among the 36 patients with KHE complicated by KMP, the male-to-female ratio was 1∶0.8; their average age was 15.0 months (range, 4.0 - 60.0 months) , and the average age of onset was 6.3 months (range, 0.8 - 48.0 months) ; 32 (88.9%) patients were diagnosed with mixed KHE, and 4 (11.1%) with deep KHE; the tumor size ranged from 2.5 cm × 4.0 cm to 20.0 cm × 24.0 cm. During the combined treatment, the average duration of prednisone administration was 6.4 weeks (range, 5.0 - 9.0 weeks) , and that of sirolimus administration was 19.3 months (range, 13.0 - 27.0 months) . After 1-5 weeks of combined treatment, platelet counts and fibrinogen levels of the 36 patients gradually returned to normal. After short-term prednisone combined with long-term sirolimus treatment, the average disease severity score decreased from 4.0 before treatment to 2.4 at 6 months and 1.6 at 12 months. After 12 months of the combined treatment, tumors mostly regressed in 32 (88.9%) patients, partially regressed in 3 (8.3%) , showed no obvious change in 1 (2.8%) . During the treatment, common adverse reactions included gastrointestinal discomfort and oral ulcers, no patient discontinued the treatment due to severe adverse reactions, and no drug toxicity-related death occurred.Conclusion:Sirolimus combined with prednisone is effective and safe for the treatment of KHE complicated by KMP.

Objective: To investigate the clinical symptoms, treatment intervention and prognosis of Kaposiform lymphangiomatosis(KLA). Methods: Medical information and clinical characteristics data of 8 KLA patients who were admitted to Department of Pediatric Surgery of West China Hospital of Sichuan University from January 2016 to February 2019 were retrospectively reviewed and analyzed. There were 5 males and 3 females with age of 5.8 years old (from 8 months to 29 years old). Results: The lesions in all patients were diffusely distributed. In all 8 patients, the lung and mediastinum were involved with different degrees. Three cases had lesions involving pelvic and abdominal organs. Three cases had lesions involving bones. One case simultaneously involved pelvic and abdominal organs, and 1 case was involved laryngeal and neck. The clinical characteristics were mainly respiratory symptoms. In the laboratory tests, 6 patients had different degrees of thrombocytopenia (minimum 3 × 10⁹/L), and 4 patients had severe fibrinogen reduction (minimum 0.42 g/L). Three patients had prolonged activated partial thromboplastin time (up to 64.2 seconds) and 3 patients had prolonged prothrombin time (up to 18.6 seconds). After surgery (including thoracotomy, chest tube, pericardiocentesis, splenectomy) and empiric medicine therapy (vincristine, sirolimus and corticosteroid), the symptoms improved in 1 case, 2 cases died of complications, 2 cases were stable and 3 cases progressed up to February 2019. Conclusions KLA is a rare disease that should be differentiated from other types of vascular diseases. Currently, there is no consensus treatment guidelines exist. Accurate diagnosis in KLA can be a challenge. The situation in patients with KLA is prone to rapid deterioration and progress. Future research efforts should seek to develop target-specific drugs for KLA.

A "general-special integrated" prevention and treatment model for chronic obstructive pulmonary disease (COPD) patients in the community was preliminarily established with the joint effets of specialists from the terital hospital and general practitioners in the community health service center.During the implementation of the model the general practitioners recieved research training and participanted in the research project of COPD management;and the "general-special integrated" outpatient clinic greatly improved the management for COPD patients.Since the establishment of the model,the number of acute attacks of COPD patients was decreased,and the proportion of standardized medication was increased.The model also improved the research ability and clinical competency of general practitioners.The established model provides experiences for the tiered-management for COPD patients in the community.

Objective To investigate the safety of trial of labor after cesarean (TOLAC) and clinical factors associated with successful TOLAC and to compare TOLAC with elective repeat caesarean section (ERCS) in terms of obstetric and neonatal outcomes.Methods A prospective cohort study was conducted among gravidas who had a history of lower segment cesarean section and were hospitalized in the Department of Obstetrics and Gynecology,the Affiliated Drum Tower Hospital of Medical School of Nanjing University from January to December 2014.Exclusion criteria included indications for caesarean section (such as placenta previa,placenta accreta,twin pregnancy,breech presentation and severe preeclampsia),serious maternal complications after cesarean section,lower uterine segment thinner than 3 mm and poor healing of uterine incision.Totally,287 gravidas were enrolled.Among them,142 chose TOLAC and the other 145 requested ERCS.Clinical data of those gravidas were collected and statistically analyzed by t-test,Log-rank test,Chi-square or Fisher's exact test.Results (1) The success rate of TOLAC was 90.8％ (129/142).There was no significant difference in maternal age,gestational age,thickness of lower uterine segment,interval between the two deliveries and neonatal birth weight and asphyxia rate between the successful (n=129) and unsuccessful (n=13) groups (all P＞0.05).Although the two groups had no significant difference in postpartum hemorrhage (PPH) rate,the gravidas who failed in TOLAC lost more blood than those who succeeded [425 (195-675) vs 200 (50-1 400) ml,P＜0.05].Moreover,higher amniotic fluid contamination rate was observed in the unsuccessful group [6/13 vs 17.1％ (22/129),P＜0.05].In the TOLAC group,99.3％ (141/142) were under continuous fetal heart rate monitoring.Incomplete uterine rupture occurred in one women without serious maternal or neonatal outcomes.The reasons for 13 failed TOLAC cases were unbearable pain during labor,abnormal labor,fetal distress and threatened rupture of uterus.(2) Compared with the ERCS group,the TOLAC group showed shorter interval from last cesarean section to the indexed delivery[5 (2-18) vs 6 (2-19) years],younger maternal age [(31±4) vs (33 ±4) years old] and less blood loss [200 (50-1 400) vs 300 (100-1 500) ml] (all P＜0.05).Conclusion Our study shows that,those who preferred TOLAC were younger,or had shorter pregnancy interval from last cesarean section.The success rate of TOLAC is high for women undergoing systematic prenatal assessment and close management during labor with less blood loss and non-serious maternal and neonatal complications compared with ERCS.

Objective To explore the influence of continuity care model on how to deal with medical waste of insulin injections among domestic patients Methods Consecutive diabetes patients using the home insulin injections were recruited as the the members of our Diabetes Education Classfrom September 2014 to March 2015. The home medical waste disposal was brought into the content of diabetes education;problems were intervened through the continuity care model. Using the questionnaire survey to investigate the cognitive and disposal of patients on medical waste, to compare the difference of results of questionnaire before and after the intervention. Results The cognitive rate of medical waste after intervention was 97.70%(212/217),while the cognitive rate before intervention was 3.69%(8/217), the difference was significant (χ2=383.63,P<0.01). Disposal of diabetes patients using the home insulin injections had improved after the continuity care, the rate of dealing with medical waste according to specification after the intervention was 91.24% (198/217),while the rate before the intervention was 3.23%(7/217), the difference was significant (χ2=337.26,P<0.01). Conclusions The continuity care model standardized the behavior of patients for disposal of medical waste, reduced the environment pollution of medical waste and decreased the spread of disease.

Objective To observe the inhibitory effect of pyridoxine hydrochloride (PN) combined with common chemotherapeutics on mice hepatoma cells H22 in vitro. Methods MTT assay was used to determine the effects of PN in combination with 10 different antineoplastic agents on H22 cells, and immuno-histochemistry was used to observe the distribution of PN in H22 cells and morphologic changes of the cells before and after PN treatment. Results After 24 hours incubation with 5 mmol/L PN, the treated cells expanded apparently with nucleus chipping. PN entered the tumor cell and was mainly condensed in cytoplasma and H22 cells were sensitive to PN. When administered concomitantly with chemotherapic agents, most of the combinations showed antagonistic effects while a few of the combinations were additive. For instance, doxorubicin (ADM) used in combination with PN inhibited cell proliferation with an IR value (IR=0.63) much lower than ADM alone (IR=0.71, P<0.01), and the CI value was less than 0.9, which indicated an antagonistic effect. However, PN in combination with ifosfamide (ICTX) showed additive effect (CI>0.9), and the IR value (IR=0.60) in combined group was higher than that (IR=0.40) in ICTX group (P<0.05). Conclusion PN treatment could increase the intracellular PLP level and result in growth inhibition and cell death, and combined administration of PN and ICTX might be a potential method to improve efficacy and to reduce toxic effects while a co-administration of PN and ADM should be avoided.

To obtain human antibodies against the Gn protein of Severe fever with thrombocytopenia syndrome virus (SFTSV) with phage display technology, this study aimed to screen anti-Gn protein antibodies from an anti-SFTSV Fab human phage display library. Antibody genes were identified by sequence analysis and the specificity of antibodies was confirmed by ELISA. The Fab antibody genes were cloned into the HL51-14 vector and expressed in a mammalian cell expression system. IgG antibodies were then purified by protein A affinity chromatography,and the results were further confirmed by ELISA,IFA,western blotting assays and micro-neutralization tests. The results showed that, after three rounds of panning, there were 390 human Fab antibodies against SFTSV particles, of which 364 were specific for nucleoprotein. Coated with the Gn protein, eight different Fab antibodies specific for Gn protein were obtained after the determination of the subtype and subclass of antibodies by gene sequencing; five of these antibodies were from the Lambda library and three were from the Kappa library. The eight IgG antibodies could specifically bind to Gn protein according to the ELISA, IFA and Western blotting assays. The micro-neutralization test showed that these eight antibodies had no neutralizing activity,but they could still provide a reference for research in human monoclonal antibodies against SFTSV.
Antibodies ; genetics ; immunology ; Bunyaviridae Infections ; genetics ; immunology ; virology ; Cell Line ; Cloning, Molecular ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Immunoglobulin G ; genetics ; immunology ; Neutralization Tests ; Phlebovirus ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

OBJECTIVE ToobservetheprotectiveeffectandmechanismofCompoundGinkgo biloba(CGB)againstalcohol-inducedliverinjury.METHODS MiceweregivenCGB0.125,0.25and 0.75 g·kg -1 ,Ginkgo biloba extract (GBE)0.1 25 g·kg -1 and bifendate(Bif)0.1 5 g·kg -1 for 8 weeks, respectively.At the end of 4th week the mice were given wine by gavage (56% V/V,0.01 L·kg -1 ), and (56% V/V,0.016 L·kg -1 )at the end of the 8th week.The serum was obtained to measure alanine transaminase (GPT),aspartate aminotransaminase (GOT),mitochondrial aspartate aminotransferase (mGOT)and tumor necrosis factor-α(TNF-α).Liver histopathology was revealed by HE staining.The protein expression of cytochrome P450 (CYP)2E1 ,NF-E2-related factor 2 (Nrf2)and TNF-αin the liverwasanalyzedbyWesternblotting.RESULTS Comparedwithnormalcontrolgroup,theactivitiesof GOT and mGOT were increased in model group (P<0.01 ).Compared with model group,CGB 0.25 and 0.75 g·kg -1 groups and Bif 0.1 5 g·kg -1 group significantly decreased the activity of GOT and mGOT in serum (P<0.05,P<0.01 ),while there was no significant difference between these groups in serum GPT activity (P>0.05).Fatty degeneration and neutrophil infiltration were significantly ameliora-ted in CGB 0.25 and 0.75 g·kg -1 groups.Preliminary mechanism research showed CGB not only increased the protein expression of Nrf2 with a positive dose-effect relationship (r=0.942,P<0.01 ), but reduced the protein expression of hepatic CYP2 E1 and the level of TNF-αin hepatic tissue with a negative dose-effect relationship (r=-0.987,P<0.05;r=-0.940,P<0.05).In addition.The level ofTNF-αwasalsosignificantlydecreasedintheserum(P<0.05,P<0.01).CONCLUSION CGB may protect the liver fro m acute alcoholic injury and the mechanis m may be that it increases the protein expression of Nrf2,restrains the protein expression of hepatic CYP2E1 and TNF-αand reduces the TNF-αlevel in the serum.