The enterotoxigenic Escherichia coli (ETEC) infection is a major factor restricting the development of animal husbandry. However, the abuse of antibiotics will lead to the antibiotic residues and emergence of antibiotic-resistant bacteria. The existing vaccines face challenges in stimulating intestinal immunity, demonstrating limited prevention effects. Therefore, it is indispensable to develop a new vaccine that is safe and suitable as a feed additive to activate intestinal immunity. This study constructed yeast-cell microcapsules (YCM) carrying the DNA vaccine against ETEC by genetic engineering. Furthermore, animal experiments were carried out to explore the regulatory effects of feeding YCM on the intestinal immune system and intestinal microbiota. Saccharomyces cerevisiae was selected as the oral delivery vehicle (microcapsules) of the DNA vaccine. The codon-optimized nucleic acid sequence of K88, the main antigen of mammal-derived ETEC, was synthesized, and the yeast shuttle vector containing the corresponding DNA vaccine expression cassette was constructed by DNA recombination. The recombinant strain of YCM was prepared by transforming JMY1. Additionally, the characteristics of the YCM strain and its feasibility as an oral vaccine were comprehensively evaluated by the fluorescence reporter assay, gastrointestinal fluid tolerance assay, intestinal epithelial cell adhesion assay, intestinal retention assessment, antiserum detection, and intestinal microbiota detection. The experimental results showed that the DNA vaccine expression cassette was expressed in mammals, and the recombinant strain of YCM could tolerate up to 8 hours of gastrointestinal fluid digestion and had good adhesion to intestinal epithelial cells. The results of mouse feeding experiments indicated that the recombinant strain of YCM could stay in the intestinal tract for at least two weeks, and the DNA vaccine expression cassette carried by YCM entered the intestinal immune system and triggered an immune response to induce the production of specific antibodies. Moreover, feeding YCM recombinant bacteria also improved the abundance of gut microbiota in mice, demonstrating a positive effect in regulating intestinal flora. In summary, we prepared the recombinant strain of YCM carrying the DNA vaccine against ETEC and comprehensively evaluated its characteristics and feasibility as an oral vaccine. Feeding the recombinant YCM could induce specific immune responses and regulate intestinal microbiota. The findings provide a reference for the immunoprevention of ETEC-related animal diseases.
Animals ; Enterotoxigenic Escherichia coli/genetics* ; Saccharomyces cerevisiae/metabolism* ; Vaccines, DNA/genetics* ; Mice ; Escherichia coli Infections/immunology* ; Escherichia coli Vaccines/genetics* ; Capsules ; Mice, Inbred BALB C ; Female

OBJECTIVE To provide reference for clinically safe drug use by mining oxaliplatin-related adverse drug events （ADE） of the nervous system. METHODS Oxaliplatin-related neurologic ADE data reported by the FDA adverse event reporting system （FAERS） between January 1st， 2004 and December 31st， 2022 were collected. The reporting odds ratio and proportional reporting ratio were used for data mining. The data were classified statistically by using systematic organ classification， high-level group term （HLGT） and preferred term （PT） in the MedDRA （version 26.0）. RESULTS A total of 7 266 cases of oxaliplatin- related ADE， which were classified as various neurological， were retrieved， and 100 PT were identified. Of these， fifty-seven PT were unspecified adverse reaction signals in the manual. Among these reports， males （46.85%） were more than females （42.98%）， the age of patients was 45-＜75 years （65.22%）， the number of reports was highest in Italy （16.32%）， and the severe type was hospitalization or prolonged hospitalization （38.16%）. The top 5 PT reports in the list of case number were peripheral neuropathy， paresthesia， neurotoxicity， loss of consciousness and dysarthria. The top 5 PT reports in the list of signal intensities were cold- induced paresthesia， neuromuscular rigidity， acute polyneuropathy， neuronal neuropathy， axonal and demyelinating polyneuropathy. A total of 13 HLGT were involved， with neurological diseases （not classified separately） having the highest number of signals （29）. CONCLUSIONS When using oxaliplatin in clinical practice， it is not only necessary to monitor the high incidence of acute and chronic peripheral neuropathy， but also to pay attention to the patient’s consciousness state and neurological symptoms. We should pay attention to the rare types of adverse reactions， such as guillain-barre syndrome， Lhermitte sign， posterior reversible encephalopathy syndrome， and hyperammoniacal encephalopathy， so as to ensure the safety of medication.

OBJECTIVE To establish a total quality management system for pharmacy intravenous admixture services （PIVAS）， in order to promote the standardization， accuracy and rationalization of clinical intravenous infusion. METHODS Based on information system in PIVAS， the management system and quality monitoring items of the whole process before， during and after PIVAS infusion preparation were formulated. The quality control and quality improvement were carried out regularly with quality management tools and methods such as PDCA （plan， do， check， process） cycle， quality control circle， and root cause analysis. The main quality control indexes of PIVAS were retrospectively analyzed before （in 2019） and after PDCA cycle management （in 2020 and 2021）. RESULTS The indexes of quality monitoring in the whole process of PIVAS infusion preparation， such as the score of drug quality management， the drug residue qualification rate and the qualified rate of drug content in infusion， were increased from 92 points， 79%， 86.4% in 2019 to 99 points， 92%， 99.8% in 2021， respectively. The indexes of safe and rational drug use， such as the ratio of intravenous irrational medical orders， the rate of drug repercussion， the rate of antibiotics use， and the rate of TCM injection use decreased from 0.98%， 6.1%， 40.55%， 39.70% to 0.23%， 3.2%， 37.18%， 26.00%， respectively. CONCLUSIONS The established total quality management system for PIVAS can improve the quality management level in the infusion preparation process， improve the quality of infusion preparation and promote clinical safe and rational drug use.

OBJECTIVE: To analyze the clinical features and prognosis of cervical adenocarcinoma (AC) and adenosquamous carcinoma of cervix (ASC). METHODS: The clinical data of 237 patients, including 201 cases of AC and 36 cases of ASC (FIGO stage ⅠB1-ⅡA), who underwent surgery in Qilu Hospital between September 2007 and September 2016 were reviewed. Clinical features of two groups were compared, and Kaplan-Meier survival analysis was performed to evaluate the prognosis. RESULTS: A larger proportion of ASC patients had lymphovascular space invasion compared with AC patients (<0.01), but no significant differences were observed in the age, FIGO stage, size of tumor, depth of stromal invasion, parametrial invasion, lymphatic metastasis and risk grade between two groups (all >0.05). The 5-year overall survival rates of AC and ASC groups were 79.4% and 78.3%, and the 5-year recurrence-free survival rates were 77.4% and 73.0%. Among patients received concurrent chemoradiotherapy, the 5-year overall survival rates were 71.0% and 61.4%, and the 5-year recurrence-free survival rates were 68.8% abd 61.1%, respectively. No significant differences were observed in 5-year overall survival rates and recurrence-free survival rates between AC and ASC patients (all >0.05). CONCLUSIONS Lymphovascular space invasion was more likely to occur in patients with ASC, but there was no significant difference in the prognosis between AC and ASC patients.
Adenocarcinoma ; diagnosis ; mortality ; Carcinoma, Adenosquamous ; diagnosis ; mortality ; Female ; Humans ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Survival Rate ; Uterine Cervical Neoplasms ; diagnosis ; mortality

Objective To explore the value of two dimentional colour Doppler flow image (2D-CDFI) combined with three-dimensional color power angiography (3D-CPA) in diagnosis of placenta accreta.Methods A total of 43 pregnant women at risk of placenta accreta selected from September 2010 to August 2015 were enrolled,and underwent 2D-CDFI and 3D-CPA to scan entire placenta.Taking the results of clinical outcome and delivery pathology of the placenta as standard,the ultrasound characteristics of 2D-CDFI and 3D-CPA were analyzed.Results Taking the results of clinical outcome and delivery pathology of the placenta as standard,24 were proved with placenta increta,3 patients with adherent placenta,2 patients with placenta percreta,14 patients with no placenta implantation.Out of 43 cases,29 cases displayed the placental thickening and rich blood vessels in placenta,and at interface of placenta and bladder wall in 2D-CDFI.For 2D-CDFI,19 cases were correctly diagnosed with placenta accrete,while 6 cases were mis-diagnosed and 4 cases missed diagnosed,the diagnosis coincidence rate by 2D-CDFI was 65.5％ (19/29).The ultrasound characteristics displayed irregular arranged myometrial arcuate artery,rich blood vessels at interface of placenta and bladder wall in 3D-CPA.For 3D-CPA,23 cases were correctly diagnosed with placenta accrete,3 cases were misdiagnosed,the diagnosis coincidence rate by 3D-CPA was 79.3％ (23/29).For 3D-CPA combined 2D-CDFI,1 case missed diagnosed,the diagnosis coincidence rate by combination 2D-CDFI with 3D-CPA was 96.6％ (28/29).Conclusions Placenta accrete can all be prenatally diagnosed by characteristic ultrasonic features of 2D-CDFI and 3D-CPA.But 3D-CPA can clearly display the range of placenta accrete lesions and the depth of the blood vessels diffused,has more advantage than two-gray scale ultrasound and 2D-CDFI and has broad application in clinic.

AIM To develop a UPLC-MS/MS method for the simultaneous content determination of adenosine,salidroside,chlorogenic acid,calycosin-7-O-β-D-glycoside,specnuezhenide,calycosin,formononetin,quercetin,apigenin and astragaloside Ⅳ in Zhenqi Fuzheng Capsules (Ligustri lucidi Fructus and Astragali Radix).METHODS The analysis of methanol extract of this drug was performed on a 40 ℃ thermostatic Inertsutain C18 column (75 mm × 3.0 mm,2 μm),with the mobile phase comprising of methanol-acetonitrile-4 mmol/L ammonium acetate flowing at 0.5 mL/min in a gradient elution manner.RESULTS Ten constituents showed good linear relationships within their own ranges (r≥0.996 0),whose average recoveries were 95.1％-104.3％ with the RSDs of less than 4.20％.CONCLUSION This sensitive and specific method can be used for the rapid quality control of Zhenqi Fuzheng Capsules.

Objective To analysis the relationship between of waist-to-height ratio(WHtR) and the non-alcoholic fatty liver disease(NAFLD).Methods A total of 7 896 were selected as observed subjects from the Kailuan Group in 2011-2012 health physical examination.A questionnaire survey,blood biochemical and abdominal ultrasound examination were finished by trained medical staff.According to previous study recommended cut points the observed subjects was divided into two groups:abdominnal obesity group(WHtR≥0.5,n=3 185) and non-abdominnal obesity group(WHtR<0.5,n=479).According to the WHtR and its quartile,the observed subjects was divided into four groups(first(WHtR<0.47),second(0.47≤WHtR<0.51),third(0.51≤WHtR<0.55) and forth(WHtR≥0.55) quartile groups).NAFLD detection rate of the four groups were Compared.Multiple Logistic regression analysis was used to analyze the influencing factors of the NAFLD.Results (1)The detection rate of NAFLD in abdominnal obesity group was higher than that in non-obesity group(62.7% vs.21.5%,P<0.01).Along with increasing WHtR level in the 4 quartile groups,the incidence of NAFLD was progressively increased,being 12.6 %,38.1%,57.5% and 72.5% respectively in the total population.After stratified by gender,15.1%,40.6%,59.6% and 75.0% in male,7.8%,31.5%,48.3% and 63.8% in female.In the same WHtR group,the NAFLD detection rate of male was higher than female,the differences were statistically significant(P<0.01).(2)Multiple logistic regression analysis showed that compared with the first quartile group,the second,third and forth quartile groups had increased risk of NAFLD after adjusting age,gender and other risk factors,the OR value being 3.19,5.93 and 9.81 in the total population.After adjusting above factors,the OR value being 3.12,6.02 and 10.45 in male,3.32,5.37 and 7.57 in female,respectively.Conclusion The incidence of NAFLD is increased along with the increasing the WHtR.

Objective To investigate the effects of waist-to-hip ratio (WHR) on the formation of carotid plaque in the middle-aged and elderly people.Methods A total of five thousand and ninety-nine participants who have taken physical examination in Tangshan Kailuan Company from June 2010 to October 2011(above 40 years old,no previous history of ischemic stroke,transient ischemic attack,myocardial infarction) were enrolled in this study.Questionnaire survey,blood biochemical and carotid artery ultrasound examination were used to examine the subjects.According to WHR,quartile division was used to divide the subjects into four groups.The detection rates of carotid plaque among the 4 groups were compared.Multivariate logistic regression analysis was used to analyze the factors effecting carotid artery plaques.Results (1) The detection rate of carotid artery plaque increased with the increase of WHR,the rates of the first,second,third and fourth group were 20.7% (571/2 753),21.9% (604/2 753),27.5% (758/2 753) and 29.8% (820/2 753) respectively.The difference was statistically significant(P=0.000).(2) Logistic regression model was used to analyze multiple factors affecting the detection rate of carotid plaque.After the correction of other factors,compared with the first group,the carotid artery plaque detection rate in the second,third,fourth group were all increased,the OR values were 1.23(95%CI:1.10~1.54),1.65(95%CI:1.52~2.13),1.67(95%CI:1.54~2.17),respectively.Conclusion The increased WHR in the middle-aged and elderly population may increase the risk of carotid plaque formation.

Background Epithelial-mesenchymal transition (EMT) is one of the pathogenesis mechanisms of posterior capule opcification (PCO).Studying EMT is of important significance for the prevention and treatment of PCO.However,EMT model is lack.Objective This study was to establish an in vitro EMT model for the study of human PCO.Methods In vitro mimic cataract enucleation was performed on 40 donor eyes,including anterior capsulorhexis,nucleus hydroexpression,and aspiration of lens fibers.The capsular bag of lens was dissected free during the surgery and pinned flat on a plastic culture dish with DMEM/F12 supplemented containing 10％ fetal bovine serum for 4 weeks.The proliferation of LECs on the capsular bag was observed by phase-contrast and dark-field microscope in 0,3,7,14 and 28 days after culture.The capsular bag tissues were collected in cultured 0,3,7 and 28 days for the preparation of sections and hematoxylin-eosin stain,and the growth and morphology of LECs were examined with optical microscope.The expression and location of α-SMA,E-cadherin and Vimentin were assessed by immunochemistry.The expression levels of α-SMA,E-cadherin and Vimentin mRNA and proteins were detected by using real-time fluorescnce quantitative PCR and Western blot in different time points.Results No LECs were seen on the uncultured capsular bag.LECs appeared in cultured day 3 on the periphery capsular bag and grew toward the center and covered the posterior capsule 7 days after cultured,with a cobblestone-like appearance.Wrinkles occurred and extended gradually along with the enhancement of bag tension.Immunochestry showed that the expression intensity of E-cadherin in the capsular bag gradually weakened,and that of α-SMA or Vimentin was gradually enhanced during the culture duration.The relative expression levels of E-cadherhin mRNA at 0,3,7,14 and 28 days after culture were 3.35±0.13,1.47±0.20,1.13±0.14,1.00±0.85 and 0.23±0.03,and relative expression levels of Vimentin mRNA were 1.00 ± 0.73,1.05 ± 0.14,2.24 ± 0.43,2.84 ± 0.34 and 8.57 ± 0.40,and those of α-SMA mRNA were 1.00 ± 0.06,2.68 ± 0.28,4.24 ± 0.05,2.05 ± 0.90 and 15.30 ± 0.19,showing significant differences among different time points (E-cadherhin mRNA:F =23.430,P =0.000;Vimentin mRNA F =8.915,P =0.002;α-SMA mRNA:F =103.500,P =0.000),with the lowest expression levels in the E-cadherhin mRNA and the highest expression levels in the Vimentin mRNA and α-SMA mRNA at 28 days during the culture period (all at P＜0.01).The gray values of E-cadherin protein expression were 1.443 ± 0.017,1.023 ± 0.003 and 0.568 ± 0.018,and those of Vimentin protein were 0.565±0.012,1.156±0.007 and 1.241±0.009,and those of α-SMA protein were 0.195±0.045,0.693±0.036 and 1.501±0.005 at 0,3 and 28 days,with significnant differences among various time points (E-cadherin:F =2 787.000,P =0.000;Vimentin:F =4 488.000,P =0.000;α-SMA:F =1 173.000,P =0.000).The expression levels were significantly declined in E-cadherhin protein and elevated in Vimentin and α-SMA proteins at 3 and 28 days after culture in comparison with before culture.Conclusions A novel in vitro EMT model of LECs is established in this study.This model can mimic a natural EMT procedure after extracapsular cataract enucleation and therefore is a useful model for the further research of the mechanism and prevention and treatment of PCO.

By taking the course of laboratory hematology in Curtin University in Australia as an ex-ample, this paper introduced the characteristics of its teaching mode, teaching contents, teaching methods, assessment methods and laboratory practice. The advantages and disadvantages of education characteristics between Australia and China were compared and analyzed on the training methods and the training objec-tives, respectively. The education characteristics of laboratory hematology in Curtin University provided an important inspiration for our education reform in the field of medical laboratory in the future.