A 68?year?old female patient was admitted to the hospital for multiple masses in the mouth and lungs as well as on dorsal hands for more than 20 days without obvious subjective symptoms. No abnormalities were found by physical examination. Dermatological examination showed two bean?sized dark?red nodules on the upper jaw as well as one pigeon egg?sized dark?red nodule on the left dorsal hand, and all the nodules were hard with smooth surfaces and limited mobility. Positron emission tomography?computed tomography (PetCT) revealed multiple metastases to the brain, lymph nodes, lungs, gastrointestinal tract, both kidneys, multiple bones and intermuscular tissues. Pathology of nodules from the upper jaw showed lowly differentiated tumor cells with osteoid matrix, chondroid structures and tumor bone in local areas, and immunohistochemical examination of tumor cells found positive staining for S100(focally), vimentin, CD99, P63 and Ki?67(60%), but negative staining for keratin. A diagnosis of osteosarcoma of the right side of the upper jaw was considered. Pathology of nodules from the dorsal hand revealed no obvious abnormalities in the epidermis, while there was a diffuse infiltration of medium?to large?sized histiocyte?like cells in the whole dermis with cell atypia and irregularly red?stained bone matrix and tumor bone in some regions. Immunopathology showed positive staining for Ki67(60%), and negative staining for CD3, CD10, CD20, Bcl?2, and Bcl?6. A diagnosis of cutaneous metastasis of osteosarcoma was made. The patient refused further treatment and died 6 months after the onset of lesions.

Objective To investigate the effect of celecoxib on the cell cycle and cell autophagy of HepG2 cell line and its possible molecular mechanism. Methods HepG2 cells were treated with celecoxib at different concentrations (0, 50, 100, 200, 500μM), then MTT was used to detect the cell proliferation, cell cycle and apoptosis were detected by flow cytometry, the cell autophagy was observed by transmission electron microscopy, and the expressions of cyclin and autophagy protein were determined using Western blot. Results Celecoxib inhibited the proliferation of HepG2 cells in a concentration dependent manner (0, 50, 100, 200 and 500 μM) and time-dependent manner (24,48h)(P<0.05). There was no significant change in necrocytosis and apoptosis after ttreated by different concentrations of (0, 50, 100, 200 and 500 μM). The celecoxib inhibited cell cycle arrest at G1 phase, the cell rate of G1 phase increased, while the cell rate of S phase decreased in a concentration dependent manner (0, 50, 100, 200 and 500 μM) and time-dependent manner (0, 8, 16, 24h)(P<0.05). The protein expressions of cyclin D1,cyclin D3, cyclin E2, CDK2 and CDK4 significantly decreased by 500 μM celecoxib (P<0.05). The celecoxib induced autophagy in HepG2 cells, and transformed autophagy protein LC3-Ⅰto LC3-Ⅱ in a concentration dependent manner. Conclusion Celecoxib can effectively inhibit the proliferation of human malignant hepatocellular carcinoma cell line HepG2, which provides a new idea for the clinical application of celecoxib.

[ ABSTRACT] AIM:To evaluate the clinical application of single nucleotide polymorphism array ( SNP array) in prenatal diagnosis for screening the abnormality of women with Down’ s syndrome ( DS) .METHODS:The amniotic fluid samples ( n=312) collected by amniocentesis for the DS screening abnormality women were tested by karyotyping and SNP array analysis, respectively.The findings of karyotyping and SNP array analysis were compared.RESULTS:Two cases of trisomy 21 were identified by karyotyping and SNP array analysis, but SNP array analysis failed to identify 6 cases of chro-mosome balanced structural rearrangement.SNP detected 176 cases copy number variants ( CNVs) in 303 cases normal karyotype were detected by SNP, including 106 benign CNVs, 61 variants of unknown significance (VOUS), 9 de novo CNVs, and none of them was pathogenic.The distribution difference of CNVs in DS screening positive group and DS screening positive plus advanced maternal age group was not statistically significant ( P>0.05) .Furthermore, we reported 14 kinds of CNVs for the first time in population.CONCLUSION:SNP array can further assure chromosome microdupli-cation/microdeletion.In normal karyotype fetus of prenatal diagnosis, SNP can detect some clinical significant CNVs.

Four genotypic mice were constructed, by ovary transplantation, with body leptin receptor (LR) + and ovary LR - in Group A, body LR + and ovary LR + in Group B, body LR - and ovary LR + in Group C, as well as body LR - and ovary LR - in Group D respectively. Mice in C and D groups showed significant differences in glucose and fat metabolism as well as ovarian function as compared with A and B groups. Leptin signal transducting system exists in the ovary. Ovarian failure was induced by hyperlipidemia and lowered gonadotropin levels, but not by defective leptin signal within ovaries in the db/db mice.

BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research.
OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s.
METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by
immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein.
RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.

Objective To evaluate the immune effects of virus-like particles ( VLPs) of VP1 pro-teins derived from norovirus GⅠ. 1 and GⅡ. 4 genotypes expressed in Hansenula polymorpha expression sys-tem. Methods SDS-PAGE and Western blot assay were performed to detect the purity of GⅠ. 1 and GⅡ. 4 VP1 proteins after purification. Morphologies of the recombinant VLPs were observed under transmission electron microscopy ( TEM) . Sizes and distributions of the VLPs were analyzed by dynamic light scattering analyzer. BT50(50% of blocking titer) was detected by HBGA (histo-blood group antigen) blocking assay in BALB/c mice immunized with different regimens. Results SDS-PAGE analysis of the purified recombinant GⅠ. 1 and GⅡ. 4 VP1 proteins showed that their purity were greater than 90%. Western blot assay con-firmed the specific bands of VLPs. TEM images showed that the sizes of purified GⅠ. 1 and GⅡ. 4 VP1 VLPs were at a mean diameter of 30-50 nm with clear border and high homogeneity, which was similar to that of wild virus. BT50 significantly increased in the groups, in which Al( OH) 3 was used as adjuvant. Con-clusion Animal studies have shown that administration of GⅠ. 1 and GⅡ. 4 VP1 VLPs in the presence of Al( OH) 3 induces detectable HBGA-blocking antibody, indicating that GⅠ. 1 and GⅡ. 4 VP1 VLPs are promising candidates for norovirus vaccine.

Cytotoxic T cells (CTLs) play a key role in the control of Hepatitis B virus (HBV) infection and viral clearance. However, most of identified CTL epitopes are derived from HBV of genotypes A and D, and few have been defined in virus of genotypes B and C which are more prevalent in Asia. As HBV core protein (HBc) is the most conservative and immunogenic component, in this study we used an overlapping 9-mer peptide pool covering HBc to screen and identify specific CTL epitopes. An unconventional HLA-A2-restricted epitope HBc141-149 was discovered and structurally characterized by crystallization analysis. The immunogenicity and anti-HBV activity were further determined in HBV and HLA-A2 transgenic mice. Finally, we show that mutations in HBc141-149 epitope are associated with viral parameters and disease progression in HBV infected patients. Our data therefore provide insights into the structure characteristics of this unconventional epitope binding to MHC-I molecules, as well as epitope specific CTL activity that orchestrate T cell response and immune evasion in HBV infected patients.
Adult ; Amino Acid Sequence ; Animals ; Binding Sites ; Epitopes ; chemistry ; immunology ; metabolism ; Female ; Genotype ; HEK293 Cells ; HLA-A2 Antigen ; metabolism ; Hepatitis B Core Antigens ; chemistry ; immunology ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Hydrogen Bonding ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Middle Aged ; Molecular Dynamics Simulation ; Mutation ; Protein Binding ; Protein Structure, Tertiary ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism

Objective To evaluate the immune effects of virus-like particles ( VLPs) assembled from the capsid protein VP1 of a recombinant norovirus ( NoV) GⅡ. 17 genotype. Methods The recombi-nant NoV GⅡ. 17 VP1 VLPs were purified, and then tested by SDS-PAGE and Western blot to analyze the purity. The size, morphology and diameter distribution of the recombinant VLPs were detected by transmis-sion electron microscopy ( TEM) and dynamic light scattering ( DLS) analyzer. The recombinant VP1 VLPs adsorbed by aluminium adjuvant were used to immunize BALB/c mice. Serum samples were collected after immunization. Specific antibody level and neutralizing antibody activity were evaluated with enzyme linked immunosorbent assay ( ELISA) and histo-blood group antigen ( HBGA)-VLP blocking test. Cross-reactivity of serum samples with GⅠ. 1 and GⅡ. 4 VP1 VLPs were detected. Moreover, cross-protection against GⅠ. 1 and GⅡ. 4 VP1 VLPs was analyzed. Results The purity of the recombinant NoV GⅡ. 17 VP1 VLP was greater than 90％ and specific bands were detected by Western blot. TEM images and DLS experiments showed that VLPs were 30-50 nm in size with good morphology and uniformity, indicating that the recombi-nant VLPs were similar to the wildtype virus. High titers of specific antibodies were detected in serum sam-ples of the immunized mice. A certain degree of cross-reactions between serum samples and VP1 VLPs of NoV GⅠ. 1 and GⅡ. 4 were observed, but no cross-protection was detected. Conclusion The recombinant GⅡ. 17 VP1 VLPs in combination with aluminum adjuvant can induce higher titers of HBGA blocking anti-bodies in mice, suggesting that it could be used as a candidate target antigen for norovirus vaccine.

Objective To evaluate the immunopotentiating effect of cyclic guanosine monophos-phate-adenosine monophosphate (cGAMP) as an adjuvant on norovirus (GⅡ. 4) virus like particles (VLPs) in the development of norovirus vaccine. Methods BALB/c mice were intramuscularly immunized with norovirus (GⅡ.4) VLPs composed of capsid protein VP1 in combination with cGAMP or Al(OH)3. Norovirus VLPs-specific antibodies in serum were detected by ELISA. A synthetic histo-blood group antigen (HBGA)-VLPs blocking assay was used to analyze neutralizing antibodies against norovirus VLPs in serum samples. Results Immunization with norovirus VLPs in the presence of cGAMP induced a strong humoral immune response in BALB/c mice. Levels of specific IgG antibodies in serum induced by using cGAMP as the adjuvant were significantly higher than those induced by using Al(OH)3adjuvant when immunization of BALB/c mice with the same dosage of VLPs. The antibody level induced by 1 μg of VLPs in combination with cGAMP was equivalent to that elicited by 10 μg of VLPs combined with Al(OH)3adjuvant. Results of the synthetic HBGA-VLPs blocking assay showed that the blocking rate in cGAMP+VLPs immunization group were significantly higher than that in Al(OH)3+VLPs immunization group when using the same dosage of VLPs. No significant difference in blocking rate was observed between cGAMP+VLPs(1 μg) and Al(OH)3+VLPs (10 μg) immunization groups. Conclusion cGAMP significantly enhanced the specific humoral immune response induced by norovirus (GⅡ.4) VLPs in mice as compared with Al(OH)3adjuvant. It might be used as a novel adjuvant to replace the traditional aluminum adjuvant in the development of norovir-us vaccine.

Objective:To systematically evaluate the prevalence of norovirus causing sporadic acute gastroenteritis in China.Methods:Relevant articles on acute gastroenteritis caused by norovirus in China published between January 2010 and October 2023 were retrieved from Wanfang, CNKI and PubMed database. The articles met inclusion and exclusion criteria were enrolled in this study. Excel software and SPSS20.0 software were used for statistical analysis. The epidemiological features of sporadic cases of acute gastroenteritis caused by norovirus in China were summarized using descriptive statistical analysis.Results:A total of 500 articles were included in this study, involving 784 486 cases of acute gastroenteritis and 670 292 samples in 32 provinces and regions. Norovirus GⅡ was the predominant genogroup causing acute gastroenteritis in China in recent years, but there were significant differences in the distribution of genotypes and epidemic strains at different times. GⅡ.4 was the predominant genotype in each year, and GⅡ.4/2006b and GⅡ.4 /Sydney_2012 were the main epidemic strains. Norovirus-related diarrhea occurred throughout the year, especially between the months of October and December. The incidence of norovirus infection was high in children under five years old and varied in different regions.Conclusions:Norovirus GⅡ was the predominant genogroup causing norovirus-related sporadic acute gastroenteritis in China, but there was an obvious genetic evolutionary trend in the epidemic strains. Factors such as epidemic strains, season and geographical region should be considered when making strategies for the prevention and control of norovirus-related diarrhea and developing vaccines.