Active immunotherapy for cancer aims to treat disease by inducing effective cellular immunity and humoral immunity.Research on virus-like particles(VLPs)vaccines has made tremendous progress in recent years,dramatically reducing morbidity and mortality from some infectious diseases.VLPs are nanoparticles self-assembled from one or more structural proteins,with highly ordered repeat sequences and good immunogenicity,which can induce strong cellular immune and humoral immune responses.VLPs can overcome immunosuppressive state of tumor microenvironment,break self-tolerance,and trigger strong cytotoxic T lymphocyte activity,which is critical for both viral clearance and destruction of cancer cells.This article mainly reviews current research progress of VLPs-based cancer vaccines and potential defects of VLPs as vaccine carriers in development of cancer vaccines.

Virus-like particles(VLPs)are nanoparticles that are self-assembled from one or more structural proteins,which can be arranged in several layers or contain a lipid outer membrane.Due to the lack of genetic material,VLPs cannot infect host cells,but are highly immunogenic and can induce immune responses different from conventional inactivated vaccines.VLPs can be produced using a variety of systems including bacterial,yeast,plant,insect and mammalian cells.Compared with traditional vaccines,VLPs have incomparable advantages,so they are becoming more and more popular in the biomedical field.To date,a series of vaccine candi-dates based on VLPs have been developed for immunization and prevention of various infectious diseases.At the same time,the recent successful application of VLPs in targeted drug delivery and gene therapy has attracted attention.This paper mainly reviews the com-monly used expression systems of VLPs and the research progress of their applications.

Objective To investigate the effects of probiotics combined with dietary intervention on pe-ripheral blood glucose and lipid metabolism indicators,placental tissue insulin signaling pathway proteins ex-pression and pregnant outcome in the patients with gestational diabetes mellitus(GDM).Methods A total of 83 patients with GDM in this hospital from December 2021 to December 2022 were selected as the study sub-jects and divided into the probiotics group(probiotics combined with diet intervention,43 cases)and control group(simple diet intervention,40 cases)by the random number table method.The levels of peripheral blood glucose,lipid and insulin resistance related indicators before the intervention and in 8 weeks after the interven-tion were compared between the two groups.The histological changes of placenta were observed by HE stai-ning.The pathological indicators were compared between the two groups.The expression levels of insulin re-ceptor substrate-1(IRS-1),glucose transporter 4(GLUT4)and synaptosome-associated protein of 23 kDa(SNAP23)in placental tissue were detected by immunohistochemistry.The pregnant adverse outcomes were compared between the two groups,and the clinical efficacy of probiotics was evaluated.Results Compared with the control group,the levels of fasting blood glucose(FBG),fasting insulin(FINS),serum triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in 8 weeks after intervention in the probiotics group were significantly decreased(P＜0.05),and the level of serum high density lipoprotein cholesterol(HDL-C)was significantly increased(P＜0.05).There were no significant differences in the incidence rates of poor villi maturation,thickening of dry villi arterioles and capillary filling in villi interstitial between the two groups(P＞0.05).Compared with the control group,the expression levels of IRS-1,GLUT4 and SNAP23 in placen-tal tissue of the probiotics group were significantly increased(P＜0.05).The incidence rates of neonatal hy-poglycemia and neonatal hyperbilirubinemia in the probiotics group were significantly lower than those in the control group(P＜0.05).Conclusion Compared with simple dietary intervention,probiotics combined with dietary intervention has more advantages in improving glucose and lipid metabolism of GDM patients,moreo-ver reduces the adverse events occurrence in newborns.

The clinical manifestations of subacute combined degeneration of spinal cord (SCD) in children are complex and vary greatly. Due to the fact that some patients with SCD may be complicated with autoimmune diseases, the high early misdiagnosis and missed diagnosis rates are observed. One case of 13-year old female with severe anemia, multiple joint swelling and pain in left limbs and paralysis of bilateral lower limbs with the extremely low level of serum vitamin B12 and poly-glandular involvement as well as a variety of positive auto-antibodies (anti-intrinsic factor antibody, anti-parietal cell antibody, thyroid peroxidase antibody, thyroid globulin antibody and perinuclear anti-neutrophil cytoplasmic antibody) was retrospectively analyzed. The patient was diagnosed as SCD with autoimmune disease (undifferentiated connective tissue disease and autoimmune polyglandular syndrome). The patient′s condition gradually alleviated after high-dose intravenous methylprednisolone, immunoglobulin, naproxen (then changed to hydroxychloroquine 1 month later), vitamin B12 and levothyroxine sodium tablets supplementation, blood transfusion and rehabilitation. SCD with autoimmune diseases is rare in children, and the clinical manifestations vary greatly. Early recognition and early treatment can improve the prognosis of SCD. The clinical data of this child were retrospectively analyzed, so as to improve the understanding of the disease by clinicians.

Objective:To investigate clinical features, risk factors and prognostic effects of paradoxical response(PR)in children with tuberculous meningitis(TBM)during anti-tuberculosis treatment.Methods:The clinical and follow-up data of TBM children admitted to the Department of Pediatrics, the Affiliated Hospital of Zunyi Medical University between January 2013 and December 2018 were retrospectively analyzed.The children were divided into the PR group and the non-PR group.Influencing factors of PR were selected by the univariate analysis, and independent risk factors were screened from these influencing factors by using the multivariate Logistic regression model.The effect of PR on long-term prognosis (≥9 months) of TBM was evaluated. Results:There were 31 cases(35.6%)with PR among the 87 TBM children enrolled, including 16 boys and 15 girls, with median age of 92(8-168)months.The median time for PR occurrence during the anti-tuberculosis treatment was 33(15-180)days.PR could present dete-rioration or recurrence of original symptoms, cerebrospinal fluid(CSF)deterioration and neuroimaging deterioration, accounting for 71.0%(22/31 cases), 80.6%(25/31 cases)and 51.6%(16/31 cases), respectively.Univariate analysis showed that stage Ⅱ, limb paralysis, cranial nerve palsy, positive tests of tuberculosis infection(T-SPOT), an increased lactate dehydrogenase(LDH)level in CSF, basilar meningeal enhancement, and tuberculosis infection outside the central nervous system were the influencing factors of the PR(all P<0.05). Multivariate analysis showed that limb paralysis, cranial nerve palsy, an increased CSF-LDH level, and positive T-SPOT were independent risk factors of PR(all P<0.05). PR was not associated with prognosis( P=0.165). Conclusions:PR occurs in 35.6% of children with TBM.Limb paralysis, cranial nerve palsy, an increased CSF-LDH level and positive T-SPOT are independent risk factors of PR.PR does not adversely affect the outcome.Identifying PR is extremely important for the prevention of some clinical misunderstandings.

The clinical features and gene mutation characteristics of 2 children who suffered from Cornelia de Lange syndrome(CdLs)and were admitted to Affiliated Hospital of Zunyi Medical University in March 2019 were retrospectively analyzed.The 2 cases developed in infancy and presented with intractable epilepsy were accompanied with developmental retardation and special appearance.It was obvious that the 2 children had SMC1A gene mutations on X chromosome.Case 1 was frameshift mutations in the SMC1A gene at c. 2561dupA(p.K854fs), and case 2 was mutations in the SMC1A gene at c. 3441+ 1G>A(exon24)splicing mutation.These were heterozygous de novo mutations, and weren′t detected in their parents, which was not reported in literatures.In this study, 2 cases of CdLs were caused by SMC1A gene mutation, which enriched the human gene mutation database.CdLs should be considered in children with early-onset epilepsy, especially appearance and developmental retardation.Genetic testing is the most important diagnostic method.

This study intends to obtain recombinant proteins of ALT1 and ALT2 isozymes by using genetic recombination technology. Monoclonal antibodies ALT1 and ALT2 with high specificity and high activity were prepared and screened (ALT1 monoclonal antibody has been successfully prepared and published). The localization, distribution and expression of ALT1 and ALT2 isozymes in human tissues were discussed. The ALT2 genes were amplified from human liver cancer cell (HepG2) by RT-PCR method. The mature ALT2 gene was subcloned into the pET32a-ALT2 prokaryotic expression vector. Its ligation product was transformed into BL21(DE3) competent cells, and transformed into competent cells to express ALT2 proteins induced by IPTG. The recombinant proteins of ALT2 were purified by nickel column (Ni⁺) affinity chromatography. Balb/c mice were immunized with recombinant proteins of ALT2. Positive serum mouse spleen cells and myeloma cells SP2/0 were selected for cell fusion. The positive cell lines were selected by indirect ELISA and subcloned by limited dilution method. Affinity chromatography was used to purify ALT2 antibodies. The expression and distribution of ALT2 in human normal tissues were detected by RT-PCR and Western blotting. Results show that the expression of ALT isoenzyme in tissues was almost the same at gene mRNA level and protein level. ALT1 is highly expressed in liver, kidney and skeletal muscle, and moderately expressed in gastrointestinal smooth muscle. ALT2 is highly expressed in fat, skeletal muscle and myocardium, and is poorly expressed in gastrointestinal smooth muscle. Immunohistochemical studies show that ALT1 is highly expressed in hepatocytes, renal medullary tubules and muscle fibers, ALT2 is highly expressed in adipocytes and myocardial cells, and ALT1 and ALT2 in gastrointestinal tissues are mainly expressed in mucosa of upper intestinal wall region. The results showed that the isoenzymes ALT1 and ALT2 were mainly expressed in the mucosa of the upper part of the intestinal wall. It is widely distributed in the tissues, providing theoretical basis for understanding the mechanism of ALT activity increase under different pathological conditions.
Alanine Transaminase ; Animals ; Cloning, Molecular ; Humans ; Isoenzymes/genetics* ; Liver ; Mice ; Recombinant Proteins

The clinical characteristics, diagnosis and treatment, and the gene mutation of 2 different phenotypes patients developed with tyrosine hydroxylase deficiency (THD) were retrospectively analyzed.Case 1 was a severe infantile parkinsonism accompanied with motor retardation, which started with psychomotor retardation without dystonia in infantile period.Clinical symptoms were fluctuating.Case 2 was a mild dopa-responsive dystonia, which started with progressive lower extremity dystonia in school age.The genetic study revealed that both patients had heterozygous mutations in tyrosine hydroxylase ( TH) gene.Case 1 was compound heterozygous mutations in the TH gene at c. 457C>T(paternal) and c. 698G>A (maternal). Case 2 was compound heterozygous mutations in the TH gene at c. 457C>T(paternal) and c. 1481C>T (maternal). Both patients dramatically improved after the treatment with Levodopa.THD should be considered in any children with or without mental retardation presenting with fluctuations symptoms or fluc-tuations dyskinesia.Genetic testing is the most important diagnostic method.

Objective:To evaluate the efficacy and safety of anti-human T lymphocyte porcine immunoglobulin (P-ATG) with recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein (rhTNFR∶Fc, Etanercept) on grade Ⅲ/Ⅳ acute graft-versus-host disease (aGVHD) after allogenic hematopoietic stem cell transplantation (allo-HSCT) .Methods:Thirty-five patients with Grade Ⅲ/Ⅳ aGVHD who received P-ATG with etanercept therapy after allo-HSCT were retrospectively analyzed. P-ATGs (5 mg·kg -1·d -1) were administrated for 3 to 5 days, and then 5mg/kg was sequentially administrated, QOD to BIW. Etanercepts were administrated 25 mg, twice a week (12.5 mg, BIW for pediatric patients) . Results:Among the 35 patients with grade Ⅲ/Ⅳ aGVHD, 21 were males and 14 females, with a median age of 10 (3-54) years. A total of 19 cases of acute myeloid leukemia, 13 of acute lymphoblastic leukemia, 1 of severe aplastic anemia, 1 of myelodysplastic syndrome, and 1 of mixed phenotypic acute leukemia were noted. The overall response (OR) rate of P-ATG with etanercept was 85.7% (30/35) , with complete response (CR) and partial response (PR) rates of 34.3% (12/35) and 51.4% (18/35) , respectively, on day 28. The OR rate of grade Ⅲ aGVHD group was higher than of grade IV aGVHD group [100% (19/19) vs. 68.8% (11/16) , P=0.004]. On day 56, the OR rate became 77.2% (27/35) , with CR and PR rates of 62.9% (22/35) and 14.3% (5/35) , respectively. The OR rate of grade Ⅲ aGVHD group was also higher than of grade Ⅳ aGVHD group [89.5% (17/19) vs. 62.5% (10/16) , P=0.009]. Thirty-five patients had no adverse effects such as fever, chills, and rash during the P-ATG infusion, and no obvious liver and kidney function damage was observed after treatment. The main treatment-related complication was infection. The reactivation rates of CMV and EBV were 77.1% (27/35) and 22.9% (8/35) , respectively, and the bacterial infection rate was 48.6% (17/35) . With a median follow-up time of 13 (1-55) months after HSCT, the 1-year and 2-year OS rates were (68.1±8.0) % and (64.3±8.4) % , respectively. The 1-year OS rate of grade Ⅲ aGVHD group was superior to grade Ⅳ aGVHD group [ (84.2±8.4) % vs. (47.6±13.1) % , χ2=3.38, P=0.05]. Conclusion:This study demonstrated that P-ATG with etanercept was effective and safe in treating grade Ⅲ-Ⅳ aGVHD after allo-HSCT.