Objective: To produce chimeric antigen receptor T cells (CAR-T) targeting human hepatocyte growth factor/c-Met (HGF/c-Met) protein and detect its cytotoxicity against non-small cell lung cancer (NSCLC) cells H1975 in vitro. Methods: The whole gene sequence of c-Met CAR containing c-Met single-chain fragment variable was synthesized and linked to lentiviral vector plasmid, plasmid electrophoresis was used to detect the correctness of target gene. HEK293 cells were transfected with plasmid and the concentrated solution of the virus particles was collected. c-Met CAR lentivirus was transfected into T cells to obtain second-generation c-Met CAR-T and the expression of CAR sequences was verified by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and western blot, and the positive rate and cell subtypes of c-Met CAR-T cells were detected by flow cytometry. The positive expression of c-Met protein in NSCLC cell line H1975 was verified by flow cytometry, and the negative expression of c-Met protein in ovarian cancer cell line A2780 was selected as the control. The cytotoxicity of c-Met CAR-T to H1975 was detected by lactate dehydrogenase (LDH) cytotoxicity assay at 1∶1, 5∶1, 10∶1 and 20∶1 of effector: target cell ratio (E∶T). Enzyme-linked immunosorbent assay (ELISA) was used to detect the release of cytokines such as TNF-α, IL-2 and IFN-γ from c-Met CAR-T co-cultured with H1975. Results: The size of band was consistent with that of designed c-Met CAR, suggesting that the c-Met CAR plasmid was successfully constructed. The results of gene sequencing were consistent with the original design sequence and lentivirus was successfully constructed. CAR molecules expression in T cells infected with lentivirus was detected by western blot and RT-qPCR, which showed c-Met CAR-T were successfully constructed. Flow cytometry results showed that the infection efficiency of c-Met CAR in T cells was over 38.4%, and the proportion of CD8(+) T cells was increased after lentivirus infection. The NSCLC cell line H1975 highly expressed c-Met while ovarian cancer cell line A2780 negatively expressed c-Met. LDH cytotoxicity assay indicated that the killing efficiency was positively correlated with the E∶T, and higher than that of control group, and the killing rate reached 51.12% when the E∶T was 20∶1. ELISA results showed that c-Met CAR-T cells released more IL-2, TNF-α and IFN-γ in target cell stimulation, but there was no statistical difference between c-Met CAR-T and T cells in the non-target group. Conclusions: Human NSCLC cell H1975 expresses high level of c-Met which can be used as a target for immunotherapy. CAR-T cells targeting c-Met have been successfully produced and have high killing effect on c-Met positive NSCLC cells in vitro.
Humans ; Female ; Receptors, Chimeric Antigen/genetics* ; Carcinoma, Non-Small-Cell Lung ; CD8-Positive T-Lymphocytes ; Interleukin-2/pharmacology* ; Tumor Necrosis Factor-alpha ; Cell Line, Tumor ; HEK293 Cells ; Lung Neoplasms ; Ovarian Neoplasms ; Immunotherapy, Adoptive

Myopia has become a major public health problem affecting the eye health of our nationals, especially adolescents, and is listed as one of the three major diseases in the world. The etiology of myopia is complex and it is currently considered to be mainly related to environmental factors and genetic factors. Dietary factors are less concerned, but it has been reported that there is a correlation between high-sugar diet and the occurrence and development of myopia. Studies have found that high-sugar diet can cause hyperinsulinemia, which mediates the increase of retinal insulin-like growth factor(IGF-1, IGF-2)expression level, and the decrease of insulin-like growth factor binding protein-3(IGFBP-3)expression level in scleral fibroblasts, thereby activating signal transduction and transcriptional activator protein 3(STAT3)signaling pathway, and then acting on scleral matrix metalloproteinases(MMP)/transforming growth factor-β(TGF-β)and other factors, resulting in a decrease in scleral extracellular matrix(ECM), eventually leading to scleral remodeling, eye axis growth and myopia. This article explores the relationship between high-sugar diet and the occurrence and development of myopia from the aspects of epidemiology and biological mechanisms by consulting relevant literature, and provides research directions for the prevention and control of myopia.

AIM: To investigate the occurrence and influencing factors of capsular contraction syndrome(CCS)after cataract phacoemulsification combined with intraocular lens implantation.METHODS: A Retrospective study was conducted on the selected 1 987 patients(1 987 eyes)undergoing cataract phacoemulsification combined with intraocular lens implantation in the hospital between September 2018 and December 2021. According to the postoperative occurrence of CCS, they were divided into CCS group and non-CCS group. The clinical data in the two groups were compared. The influencing factors of CCS were analyzed by multivariate Logistic regression analysis. And the predictive model was constructed.RESULTS: There were 38 eyes with postoperative CCS among the 1 987 cataract patients(1 987 eyes), with an incidence of 1.91%. The proportions of cases with age ≥65 years, diabetes mellitus, glaucoma, retinitis pigmentosa, uveitis and hydrophilic intraocular lens in CCS group were significantly higher than those in the non-CCS group(all P<0.05). Multivariate Logistic stepwise forward regression analysis showed that age ≥65 years, diabetes mellitus, retinitis pigmentosa, uveitis and hydrophilic intraocular lens were risk factors of CCS after phacoemulsification combined with intraocular lens implantation(P<0.05). The predictive model constructed based on regression coefficients of the risk factors had good goodness of fit(P=0.421).CONCLUSION: Advanced age, diabetes mellitus, retinitis pigmentosa, uveitis and material properties of intraocular lens are important influencing factors of postoperative CCS.

This study explored the correlation between color and chemical components of Chrysanthemi Indici Flos(CIF), aiming at providing a reference for its procurement, evaluation, and breeding. Colorimeter and ultra-performance liquid chromatograph(UPLC) were used to determine the color(lightness-shade chromaticity value L~*, red-green chromaticity value a~*, yellow-blue chromati-city value b~*) and chemical components(cynaroside, linarin, luteolin, apigenin, and chlorogenic acid) of 84 CIF germplasms, respectively. Diversity analysis, correlation analysis, regression analysis, and cluster analysis were performed. The results showed that the color and chemical components of CIF were diversified. Chlorogenic acid was in significantly positive correlation with L~* and b~* and significantly negative correlation with a~*. Cynaroside and grey relational grade γ_i of chemical components were in significantly po-sitive correlation with b~* and L~*, respectively, whereas linarin, luteolin, and apigenin had no significant correlation with L~*, a~*, or b~*. The 84 CIF germplasms were clustered into 4 clades. In addition, germplasms in clade Ⅲ had higher γ_i and total color value(E~*_(ab)) than those in other clades, with the best quality and color, and a germplasm with the highest quality, bright yellow color, and highest content of linarin was screened out in this clade. Thus, CIF with bright yellow color had high content of cymaroside and chlorogenic acid and thereby high quality. In summary, the color can be used to quickly predict the quality of CIF. Our results provided data for the evaluation of CIF quality by color and a reference for its procurement and breeding.
Chrysanthemum/chemistry* ; Luteolin/analysis* ; Chlorogenic Acid/analysis* ; Apigenin/analysis* ; Plant Breeding ; Excipients ; Chromatography, High Pressure Liquid/methods*

Objective: To investigate the clinical features of children with chronic nonbacterial osteomyelitis (CNO), and raise awareness among clinicians. Methods: In this retrospective study, 18 patients with CNO who were diagnosed in Children's Hospital of Fudan University from January 2015 to December 2021 were included. Results: Eighteen children with CNO (12 males, 6 females) were identified. Their age at onset was 9 (5, 11) years, the delay in diagnosis was 2 (1, 6) months, and follow-up-was 17 (8, 34) months. The most common symptoms were fever in 14 children, as well as bone pain and (or) arthralgia in 14 children. In terms of laboratory results, normal white blood cell counts were observed at onset in 17 patients; increased erythrocyte sedimentation rate (ESR) in all patients; increased C reactive protein (CRP) over the normal value in 14 patients. Of the 18 patients, 2 had positive antinuclear antibodies, while none had positive human leukocyte antigen-B27 or rheumatoid factor. Imaging examination revealed that all the patients had symmetrical and multifocal skeletal lesions. The number of structural lesions detected by imaging investigation was 8 (6, 11). The most frequently affected bones were tibia in 18 patients and femur in 17 patients. Bone biopsy was conducted in 14 patients and acute or chronic osteomyelitis manifested with inflammatory cells infiltration were detected. Magnetic resonance imaging (MRI) found bone lesions in all the patients and bone scintigraphy were positive in 13 patients. All the patients were treated with nonsteroidal anti-inflammatory drugs, among whom 10 cases also treated with oral glucocorticoids, 9 cases with traditional disease modifying anti-rheumatic drugs, 8 cases with bisphosphonates and 6 cases with tumor necrosis factor inhibitors. The pediatric chronic nonbacterial osteomyelitis disease activity score, increased by 70% or more in 13 patients within the initial 6-month follow-up. Conclusions: The clinical manifestations of CNO are lack of specificity. The first symptom of CNO is fever, with or without bone pain and (or) arthralgia, with normal peripheral blood leukocytes, elevated CRP and (or) ESR. Whole body bone scanning combined with MRI can early detect osteomyelitis at subclinical sites, and improve the diagnostic rate of CNO.
Female ; Male ; Humans ; Child ; Retrospective Studies ; Osteomyelitis/drug therapy* ; Arthralgia ; Diphosphonates ; Fever ; Graft vs Host Disease

Objective:To explore the potential targets and mechanism of action of "Clematis Radix et Rhizoma-Trichosanthis Radix" based on network pharmacology. Method:Chinese Medicine System Pharmacology Analysis Platform（TCMSP） was used to screen out active ingredients and corresponding target proteins of Clematis Radix et Rhizoma and Trichosanthis Radix according to oral bioavailability（OB） and drug likeness（DL），cancer disease targets were screened out using GeneCards and OMIM databases，R language software was used to screen out common targets of clematis，trichosanthin and cancer diseases， Cytoscape 3.7.2 software was used to construct a network map of "drug-active ingredient-disease-target"， STRING database was used to draw protein protein interaction（PPI）of common target proteins， R language software was used to perform enrichment analysis of gene ontology（GO） functions and Kyoto encyclopedia of genes and genomes（KEGG） channels on effective targets. Result:A total of 9 effective active ingredients were obtained from Clematis Radix et Rhizoma-Trichosanthis Radix powder pair. A total of 31 target genes were searched，and 814 relevant target genes were searched from cancer diseases. The two kinds of relevant target genes were matched to obtain 9 common target genes，which mainly involved endopeptidase，cysteine-type endopeptidase activities involving in the apoptosis process and cancer necrosis factor receptor superfamily binding and other biological processes，and played a role in the treatment of cancers by regulating apoptosis，measles，hepatitis B，kaposi sarcoma-associated herpesvirus infection，p53，interleukin-17（IL-17），tumor necrosis factor（TNF） and many other pathways. Conclusion:The mechanism of Clematis Radix et Rhizoma-Trichosanthis Radix in the treatment of cancer is preliminarily studied. Clematis Radix et Rhizoma-Trichosanthis Radix has multiple active ingredients and can play a role in treating cancer through multiple targets and multiple pathways.

Objectives: To evaluate the immunogenicity of Methods: Protein extracts from Results: Immunization with Conclusion This is the advanced study to investigate the immunogenicity of
Animals ; Antibodies, Bacterial/immunology* ; Antigens, Bacterial/immunology* ; Bacterial Proteins/immunology* ; Cross Reactions ; Cytokines/immunology* ; Female ; Genome, Bacterial ; Immunoglobulin G/immunology* ; Immunoglobulin M/immunology* ; Macrophages/immunology* ; Mice, Inbred BALB C ; Mycobacterium avium Complex/immunology* ; Mycobacterium tuberculosis/immunology* ; Tuberculosis Vaccines/administration & dosage* ; Whole Genome Sequencing

There are 16 species and 7 varieties of medicinal plants of Trollius in China, with effects in clearing heat, detoxification and swelling, they are used for the treatment of acute and chronic tonsillitis, acute otitis media, chronic bronchitis and urinary tract infections. Through retrieval of literatures on nasturtium from 1972 to 2019 in a number of databases, such as CNKI, Wanfang Database, Baidu Academic and PubMed, the cultivation and quality control methods, extraction and purification processes, chemical composition, pharmacological effects, pharmacokinetics and metabolism of medicinal plants of the genus nasturtium were summarized. The researches of cultivation mainly focus on the establishment of seed treatment and tissue culture system. Quality control is mainly based on orientin and flavonoids. Research on extraction and purification technology mainly focused on the extraction and purification of total flavonoids. In terms of chemical composition, there are currently more than 100 major compounds isolated from this genus, including flavonoids, organic acids, alkaloids, coumarins and styrenes. Among them, there are 80 flavonoids, 12 organic acid monomers, 3 alkaloid monomers, 4 coumarin monomers and 14 styrene monomers. In terms of pharmacological action, there are mainly in vitro experiments of crude extracts of medicinal materials. The data of pharmacokinetics and metabolism mainly focus on the study of flavone monomers.

Objective: To observe the clinical efficacy of mind-refreshing and balance-restoring needling method combined with Frenkel exercises in treating ataxia after cerebral stroke. Methods: The recruited 120 patients were randomized into an observation group and a control group, with 60 cases in each group. The control group was intervened by mind-refreshing and balance-restoring needling method, while the observation group was given additional lower-limb Frenkel exercises. Before and after treatment and at the follow-up, the ataxic lower-limb function was scored using Berg balance scale (BBS) and international cooperative ataxia rating scale (ICARS), and Barthel index (BI) was adopted to score the activities of daily living (ADL). Results: After treatment, the markedly effective rate was 70.2％ and the total effective rate was 96.5％ in the observation group, versus 39.7％ and 87.9％ in the control group, and the differences in the markedly effective rate and the total effective rate were statistically significant (P<0.01, P<0.05). The intra-group comparisons showed that the BBS, ICARS and BI scores after treatment and at the follow-up were significantly different from those before treatment in both groups (all P<0.01).There were significant differences in the BBS score between the two groups after treatment and at the follow-up (P<0.05, P<0.01); the between-group differences in the ICARS and BI scores were statistically insignificant after treatment (both P>0.05), while the between-group differences in the ICARS and BI scores were statistically significant at the follow-up (both P<0.05). The interaction effects between the scoring time of BBS and BI and the group factor were statistically significant (P<0.01, P<0.05). Conclusion: Mind-refreshing and balance-restoring needling can effectively improve the lower-limb ataxic symptoms and ADL after stroke; when combined with Fenkel exercises, this needling method can produce more significant efficacy.

OBJECTIVE: To study the single nucleotide polymorphisms (SNPs) in promoter region of the Jk gene and its allele frequency as well as distribution characteristics in the Chinese Han nationality population. METHODS: 127 blood samples containing 8 Jk(a-b-) and 119 samples (as control) taken randomly from voluntary blood donors of Chinese Han nationality persons in Shenzhen Blood Center were collected. The Kidd phenotypes were identified by using the serologic test and urea hemolysis test; the Jk promoter, exon 1-11 region and respective flanking area were amplified and sequenced, then the sequence information was analyzed. RESULTS: 8 Jk(a-b-) samples all carried JkB/JkB allele which belongs to 2 kind of Jk genotypes commonly observed in Chinese Han nationality population. 6 IVS5-1g>a and 2 896G>A were found in 8 Jk(a-b-) samples. Besides, all Jk(a-b-) samples were homozygous for JkB/JkB allele. Three SNPs-110(rs900974), -160(rs1484877) and -258(rs1484878) in promoter region of the Jk gene were found and sequenceds calculation of allele and genotype frequencies showed that the result accorded with Hardy-Weinberg equilibrium, indicating that the population in this study possesses representative characteristics of the Chinese Han nationality population. CONCLUSION The polymorphism of the Jk gene occurs in promoter region. This study calculates the allele frequencies of three SNPs-110(rs900974), -160(rs1484877) and -258(rs1484878) in promoter region of the Jk gene, and shows their distribution characteristics in distinct Kidd phenotypes. These findings provide the basic foundation for further population genetics research.