Toxoplasma gondii is an obligatory intracellular parasite which infects a variety of warm-blooded animals and causes toxoplasmosis. Toxoplasmosis seriously endangers human health and animal husbandry production. As one of the effective gene editing tools, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system has been widely used for knockout of genes in T. gondii. This review summarizes the applications of the CRISPR/Cas9 technology in vaccines against single- and double-gene deletion strains of T. gondii, so as to provide insights into development of toxoplasmosis vaccines.

Objective:To further investigate the safety and efficacy of Belimumab in the treatment of patients with systemic lupus erythematosus (SLE).Methods:All SLE patients treated with Belimumab from May 1, 2020 to February 1, 2022 in Peking Union Medical College Hospital were retrospectively collected and analyzed. The clinical manifestations, the Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2000) score, and laboratory test such as levels of anti-dsDNA antibody, the medication before and after Belimumab treatment, adverse events were collected. Normally distributed data were tested using the t-test, otherwise the Wilcoxon paired signed rank test was used. Results:A total of 81 patients were enrolled in this study. The use of belimumab could significantly decrease the SLEDAI-2000 score [10.00(7.75, 12.00) vs. 4.00(3.75, 6.00), Z=-5.38, P<0.001], ESR of SLE patients [19.50(12.75, 32.25) mm/1 h vs. 14.00(7.75, 20.25) mm/1 h, Z=-3.71, P=0.003], anti-dsDNA titer detected by CLIFT [300.00 (117.00, 864.00) vs. 183.00(100.00, 471.00), Z=-4.15, P=0.001], meanwhile, increase the complement C3 [0.78 (0.62, 0.97)g/L vs. 0.69 (0.55, 0.84)g/L, Z=-4.68, P<0.001], and the complement C4 [0.12 (0.08, 0.19)g/L vs. 0.10 (0.05, 0.14)g/L, Z=-4.78, P<0.001]. We also observed that with the use of Belimumab, the dosage of Glucocorticoids decreased significantly, which were [10.00(7.50, 22.50) mg vs. 7.50(5.00, 10.00) mg, Z=-4.90, P<0.001]. In addition, the antibody of IgG, IgA and IgM decreased significantly. Only one patient stopped the administration of Belimumab due to the low level of immunoglobulin. Conclusion:Belimumab can alleviate disease activity of patients with SLE and help in safely tapering the daily dose of glucocorticoid with good safety.

Yeast autolysis affects the flavor and quality of beer. The regulation of yeast autolysis is a need for industrial beer production. Previous studies on brewer's yeast autolysis showed that the citric acid cycle-related genes had a great influence on yeast autolysis. To explore the contribution of isocitrate dehydrogenase genes in autolysis, the IDP1 and IDP2 genes were destroyed or overexpressed in typical lager yeast Pilsner. The destruction of IDP1 gene improved the anti-autolytic ability of yeast, and the anti-autolytic index after 96 h autolysis was 8.40, 1.5 times higher than that of the original strain. The destruction of IDP1 gene increased the supply of nicotinamide adenine dinucleotide phosphate (NADPH) and the NADPH/NADP⁺ ratio was 1.94. After fermentation, intracellular ATP level was 1.8 times higher than that of the original strain, while reactive oxygen species (ROS) was reduced by 10%. The destruction of IDP2 gene resulted in rapid autolysis and a decrease in the supply of NADPH. Anti-autolytic index after 96 h autolysis was 4.03 and the NADPH/NADP⁺ ratio was 0.89. After fermentation, intracellular ATP level was reduced by 8% compared with original strain, ROS was 1.3 times higher than that of the original strain. The results may help understand the regulation mechanism of citric acid cycle-related genes on yeast autolysis and provide a basis for the selection of excellent yeast with controllable anti-autolytic performance.
Humans ; Isocitrate Dehydrogenase/genetics* ; NADP ; Reactive Oxygen Species ; Autolysis ; Adenosine Triphosphate

Objective:To analyze the application effect of "Internet+" early grading follow-up management mode after enterostomy for colorectal neoplasms.Methods:The 186 patients who underwent enterostomy for colorectal neoplasms in Lishui Central Hospital were selected as the research objects. The patients enrolled from January 2019 to February 2020 served as the control group, and the patients enrolled from March 2020 to April 2021 served as the observation group, with 93 patients in each group. Routine follow-up measures were taken in the control group, while "Internet+" was used in the observation group for early grading follow-up management of patients with enterostomy for colorectal neoplasms. The timely follow-up feedback, score of skin condition around stoma, incidence of stoma complications and nursing satisfaction during follow-up were compared between the two groups.Results:The two-day non feedback rate in the control group was 20.43% (19/93) and that in the observation group was 7.53% (7/93), the difference was statistically significant ( χ2 = 6.44, P<0.05) .The score of skin condition around stoma in the observation group was (2.53 ± 0.78) points 3 months after discharge, which was significantly better than that in the control group 4.02 ± 1.13, the difference was statistically significant ( t = 10.47, P<0.05). The incidence of stoma complications in the observation group was 9.68% (9/93) , which was lower than that in the control group 21.51% (20/93) , the difference was statistically significant ( χ2 = 4.94, P<0.05). The nursing satisfaction in the observation group was 98.92% (92/93) , which was significantly higher than that in the control group 91.40% (85/93) , the difference was statistically significant ( χ2 = 4.20, P<0.05). Conclusions:"Internet+" early grading follow-up management model can significantly improve the follow-up effect after enterostomy for colorectal neoplasms, improve the skin condition around stoma, reduce stoma complications and improve nursing satisfaction.

Objective:To explore the effect of progressive focusing-based interview combined with standardized stoma nursing in patients with enterostomy.Methods:From June 2019 to June 2021, 150 patients with enterostomy admitted to Lishui Central Hospital were selected by convenience sampling method as the research objects. According to the random number table method, the patients were divided into the control group and the observation group, 75 cases in each group. The control group was given routine psychological nursing combined with standardized stoma nursing, and the observation group was implemented with progressive focusing-based interview combined with standardized stoma nursing, and the intervention period was two months. The stigma, disease coping style and quality of life of the two groups of patients before and after nursing were compared.Results:After two months of nursing, the scores of each dimension of the Social Impact Scale in the observation group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . The confrontation dimension score of the Medical Coping Modes Questionnaire (MCMQ) in the observation group were higher than those in the control group, and the avoidance and resignation dimension scores were lower than those in the control group, with statistical differences ( P<0.05) . The scores of each dimension of the Stoma Quality of Life in the observation group were higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Progressive focusing-based interview combined with standardized stoma nursing can reduce disease stigma in patients with enterostomy, improve their coping styles and quality of life.

OBJECTIVE: To analyze the clinical features and genetic basis of three children with mental retardation, language impairment and autistic features due to de novo variants of FOXP1 gene. METHODS: Clinical data of the children were collected.Trio-whole exome sequencing was carried out for the children and their parents. Pathogenicity of the variants was analyzed through bioinformatics prediction. RESULTS: All of the children had various degrees of mental retardation in conjunct with language deficit, global developmental delay, abnormal behavior and peculiar facial features, among whom two also developed autism spectrum disorders. The results of genetic testing showed that all three children harbored de novo variants of the FOXP1 gene, namely c.613_c.614delCTinsTA, c.1248delC and c.1393A>G. Two of these were frameshift variants and one was missense variant, which were all rated as pathogenic based on the guidelines of the American College of Medical Genetics (ACMG). Database search suggested that c.613_c.614delCTinsTA and c.1248delC were unreported previously. CONCLUSION For the three children from unrelated families with mental retardation in conjunct with language deficit, global growth delay, abnormal behavior and peculiar facial features, the c.613_ c. 614delCTinsTA, c.1248delC and c.1393A>G variants of the FOXP1 gene may be the pathogenic factors. Above cases have further expanded the genotype-phenotype profile of FOXP1 deficiency syndrome.
Autistic Disorder/genetics* ; Child ; Forkhead Transcription Factors/genetics* ; Genetic Testing ; Humans ; Intellectual Disability/genetics* ; Language Development Disorders/genetics* ; Repressor Proteins/genetics* ; Whole Exome Sequencing

Objective:To explore the efficacy and safety of anti-tumor necrosis factor alpha (TNFα) monoclonal antibodies (mAbs) for severe/refractory vasculo-Behcet′s disease (BD).Method:The clinical data of severe/refractory vasculo-BD patients treated with anti-TNFα mAbs were retrospectively analyzed. Response of anti TNFα mAbs was analyzed. The dosage changes of glucocorticoid, the level of erythrocyte sedimentation rate (ESR) and hypersensitive C-reactive protein (hsCRP) before and after treatment were recorded, as well as side effects.Result:Sixteen patients were enrolled. Arterial lesions were reported in 12 patients, including 9 with arterial aneurysm, 6 with arterial dilation, 2 with stenosis and 2 with occlusion. Seven patients presented venous thrombosis, including lower extremity veins ( n=6), cerebral venous sinus ( n=2) and inferior vena cava system ( n=2). Two cases had both arterial and venous involvement. Before the application of TNFα mAbs, all 16 patients failed to response to prednisone or its equivalent dose of 40 (7.5-90) mg/d in combination with cyclophosphamide, methotrexate, thalidomide or azathioprine for median 4 (0-156) months. After a mean duration of treatment for (17.1±6.5) months, 15 patients achieved complete remission and 1 patient achieved partial remission. Three patients received surgery without any postoperative complications. After using anti TNFα mAbs, the dosage of prednisone [5(0-12.5)mg/d vs. 40(7.5-90)mg/d, P<0.01], ESR [(7.3±4.6) mm/1h vs. (33.5±26.7) mm/1h, P<0.01] and hsCRP [1.9(0.2-11.4) mg/L vs. 24.3(0.4-113.9) mg/L, P<0.01] were significantly decreased. Side effects were observed in 2 patients. One developed pulmonary infection 12 months after adalimumab with conventional treatment. Another patient had allergy to infliximab then switched to adalimumab. Conclusion:In combination with corticosteroids and immunosuppressants, anti-TNF α mAbs are effective and well-tolerated in severe/refractory vasculo-BD, with a favorable steroid -sparing effect and rare postoperative complications.

Objective To explore the use of biological agents in Neuro-Beh(c)et's disease (NBD).Methods We retrospectively reviewed the clinical data of five NBD patients treated with biological agents who were in-patients at Peking Union Medical College Hospital between June 2009 and June 2016.The continuous variables were analyzed by t test.Results All five cases (4 male and 1 female) were severe and/or refractory patients with parenchymal involvement (pNBD).Their age at neurological onset was (31±12) years old.Four cases presented with multiple lesions.The brainstem,spinal cord,cerebral hemisphere and cerebellum involvement were presented in 4,3,3 and 2 patients,respectively.The Rankin score at the onset of NBD was (4.0±0.7).The biological agents were administrated when corticosteroids and immunosuppressant were ineffective.Three cases received tumor necrosis factor (TNF)-α inhibitors therapy,among whom one patient had gastrointestinal ulceration.One patient with refractory retinal vasculitis received interferon-α therapy.One patient treated with tocilizumab [interleukin (IL)-6R inhibitor] had high level IL-6 in the cerebrospinal fluid.All patients achievcd clinical improvements and the Rankin score significantly decreased to (2.2±0.8) when compared with the baseline (t=4.81,P＜0.01) after the treatment with biological agents.The corticosteroid dose was tapered in all cases and the numbers of immunosuppressants were reduced in most cases,indicating a potential steroid and immunosup-pressant-sparing effect.No serious adverse events were observed during the follow-up.Conclusion Neurological involvement is a severe complication of Behqet's disease.We can take appropriate biological agents such as TNF-α inhibitors or interferon-α into consideration when patients have severe/refractory pNBD.

Aged ; Anesthesia ; methods ; Electrocardiography ; Heart Ventricles ; Hip Fractures ; surgery ; Humans ; Intraoperative Complications ; etiology ; Male ; Myocardial Infarction ; etiology

Objective To investigate the pre-emptive analgesia effects of hydromorphone on stress reaction in patients undergoing gynecological laparoscopic surgery.Methods Forty ASA Ⅰ or Ⅱ patients [age:45-58 years,body mass index:18-24 kg· (m2)-1] undergoing gynecological laparoscopic surgery were randomly divided into 2 groups (n =20 for each group):treatment group and control group.Hydromorphone (1 mg) was intravenously injected before anesthesia in treatment group.In the two groups,after routine induction and incubation,remifentanil (0.2 μg· min-1 · kg-1) and propofol (0.1 mg· min-1· kg-1) were injected with micro perfusion pump,cisatracurium was injected intermittently.Injection of remifentanil and propofol was stopped when skin suture started.The concentrations of epinephrine(E) and norepinephrine(NE) were obtained before induction (t1),pneumoperitoneum (t2),1 h after pneumoperitoneum (t3) and extubation (t4),respectively.The heart rate,blood pressure and the time of operation to extubation of the patients were recorded.Results There were no significant changes in extubation time after operation among the groups.Heart rate and systolic blood pressure were significantly lower in the treatment group than in the control group (P ＜ 0.05).Plasma E and NE levels at t1 were significantly lower than those at t2,t3 and t4.Plasma E and NE were significantly lower in treatment group than that of control group at t2,t3 and t4 (P ＜ 0.05).Conclusion Pre-emptive analgesia of hydromorphone can significantly decrease the release of epinephrine and norepinephrine during and after laparoscopic surgery,restrain the increase of heart rate and systolic blood pressure during extubation,without influence of revival time.