BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients ...BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients were usually affected by heterozygous dominant mutations in the chloride voltage-gated channel 7(ClCN7)gene and present early-onset osteoarthritis or recurrent fractures.In this study,we report a case of persistent joint pain without bone injury or underlying history.CASE SUMMARY We report a 53-year-old female with joint pain who was accidentally diagnosed with ADO-II.The clinical diagnosis was based on increased bone density and typical radiographic features.Two heterozygous mutations in the ClCN7 and Tcell immune regulator 1(TCIRG1)genes by whole exome sequencing were identified in the patient and her daughter.The missense mutation(c.857G>A)occurred in the CLCN7 gene p.R286Q,which is highly conserved across species.The TCIRG1 gene point mutation(c.714-20G>A)in intron 7(near the splicing site of exon 7)had no effect on subsequent transcription.CONCLUSION This ADO-II case had a pathogenic CLCN7 mutation and late onset without the usual clinical symptoms.For the diagnosis and assessment of the prognosis for osteopetrosis,genetic analysis is advised.展开更多
AIMS To clarify whether the lower interferon gamma (IFNγ) production by lymphocytes in patients with liver diseases is due to defects of lymphocytes themselves or of other cofactors such as interleukin-2(IL-2). METHO...AIMS To clarify whether the lower interferon gamma (IFNγ) production by lymphocytes in patients with liver diseases is due to defects of lymphocytes themselves or of other cofactors such as interleukin-2(IL-2). METHODS Peripheral blood mononuclear cells (PBMCs) from patients with various liver diseases were cultured with or without PHA and IL-2. The cells were harvested and counted and the su- pernatants were tested for IFNγ by a sensitive and quantitative ABC-ELISA. RESULTS IFNγ was not round in serum samples from patients as well as normal individuals. However,in supernatants of non-in- duced and induced PBMCs,IFN7 was detected by ABC-ELISA. In non-induced PBMCs (group 1),the content of IFNγ in super- natants from control,CAH,CPH and HCC was 8.72 μg/L, 5.03 μg/L,6.02 μg/L and 4.91 μg/L respectively. The pro- duction of IFNγ in liver disease was significantly decreased,com- pared to control. In group 2 in which PBMCs were stimulated with PHA,the content of IFNγ was 22.71,17.12,14.54 and 17.63 μg/L respectively. In group 3 in which PBMCs were in- duced by IL-2,the amount of IFN7 in supernatant from control (60.67 μg/L) was much larger than those from CAH (21.70 μg/ L),CPH (24.00 μg/L) and HCC (19.15 μg/L) (P<0.01). Comparing the amount of IFNγ in group 3 (IL-2-induced) with that in group 1 (non-induced),we found that IFNγ production was en- hanced by nearly 4 folds in liver diseases and by over 7 folds in control,Whereas the number of PBMCs,whether from liver dis- eases or from control,was increased by only approximately 3 folds. CONCLUSIONS The decreased production of IFNγ in liver dis- eases including HCC is mainly due to endogenous defects of lym- phocytes though the defects of stimulating cofactors such as IL-2 may also be involved.展开更多
AIM To investigate the therapeutic potential of gamma interferon (IFN γ) gene modified human hepatocellular carcinoma (HCC) cells. METHODS The IFN γ gene was introduced retrovirally into four HCC cell lines. S...AIM To investigate the therapeutic potential of gamma interferon (IFN γ) gene modified human hepatocellular carcinoma (HCC) cells. METHODS The IFN γ gene was introduced retrovirally into four HCC cell lines. Secreted IFN γ activity was assessed using bioassay. The expression of MHC molecules was detected by FACS. Tumorigenicity was analysed by tumor formation in nude mice. RESULTS Four IFN γ gene transduced HCC cell lines secreted different amounts of IFN γ, as in the same case of five clones derived from one HCC cell line. Transduction with IFN γ caused significant increase in the expression of major histocompatibility complex (MHC) antigens on HCC cells. The expression of HLA class Ⅰ was increased by 2-3 times in terms of mean fluorescence intensities, while for class Ⅱ expression, the percentage of positive cells augmented from <10% to >50%. When equal amount of tumor cells were injected into nude mice, the tumorigenicity of some transduced cells decreased dramatically. CONCLUSION IFN γ gene transduction can convert weakly immunogenic HCC cells to activate antitumor immune response, and further pave the way for the future use of such gene modified tumor cells as a modality for the cancer immunotherapy.展开更多
BACKGROUND： The supernatant of interferon-gamma （IFNγ） co-cultured with neonatal rat cortical glia can promote the cells in embryonic basal forebrain/septal nuclei to differentiate into cholinergic neurons, but th...BACKGROUND： The supernatant of interferon-gamma （IFNγ） co-cultured with neonatal rat cortical glia can promote the cells in embryonic basal forebrain/septal nuclei to differentiate into cholinergic neurons, but the mechanism is still unclear. OBJECTIVE： To analyze the pathways for IFNγ to promote the differentiation of primarily cultured cholinergic neurons in rat embryonic basal forebrain/septal nuclei through culture in different conditioned medium. DESIGN： A controlled experiment taking cells as the observational target. SETTINGS： Department of Biochemistry and Molecular Biology, Youjiang Medical College for Nationalities; Department of Cell Biology, Beijing University Health Science Center. MATERIALS： Sixty-four pregnant Wistar rats for 16 days （250-350 g） and 84 Wistar rats （either male or female, 5-7 g） of 0-1 day after birth were provided by the experimental animal department of Beijing University Health Science Center. Rat IFNγ were provided by Gibco Company; Glial fibrillary acidic protein by Huamei Company. METHODS： The experiments were carried out in the Department of Cell Biology, Beijing University Health Science Center and Daheng Image Company of Chinese Academy of Science from July 1995 to December 2002. ① Interventions： The nerve cells in the basal forebrain/septal nuclei of the pregnant Wistar rats for 16 days were primarily cultured, and then divided into four groups： Blank control group （not any supernatant and medium was added）; Control group （added by mixed glial cell or astrocyte conditioned medium）; IFNγ group （added by mixed glial cell or astrocyte conditioned medium＋IFNγ）. Antibody group （added by mixed glial cell or astrocyte conditioned medium＋IFNγ＋Ab-IFNγ）. Mixed glial cell or astrocyte conditioned medium was prepared using cerebral cortex of Wistar rats of 0-1 day after birth. ② Evaluation： The immunohistochemical method was used to perform the choline acetyltransferase （ChAT） staining of cholinergic neurons. The ChAT positive cells were counted. MAIN OUTCOME MEASURES： Comparison of ChAT positive cells in rat basal forebrain and septal nuclei in different conditioned medium. RESULTS： ① ChAT positive cells in mixed glial cell conditioned medium： The ChAT positive cells in the IFNγ group and antibody group were significantly more than those in the control group （P 〈 0.01）. ② ChAT positive cells in astrocyte conditioned medium： The ChAT positive cells in the IFNγ group were significantly more than those in the control group, but there was no significant difference between the antibody group and control group （P 〉 0.05）. CONCLUSION： IFNγ cannot directly promote the differentiation of cholinergic neurons, but plays a role through activating glial cells （except astrocytes） to produce IFNγ like molecules.展开更多
基金Supported by the Science and Technology Plan Program of Sichuan of China,No.2018JY0608。
文摘BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients were usually affected by heterozygous dominant mutations in the chloride voltage-gated channel 7(ClCN7)gene and present early-onset osteoarthritis or recurrent fractures.In this study,we report a case of persistent joint pain without bone injury or underlying history.CASE SUMMARY We report a 53-year-old female with joint pain who was accidentally diagnosed with ADO-II.The clinical diagnosis was based on increased bone density and typical radiographic features.Two heterozygous mutations in the ClCN7 and Tcell immune regulator 1(TCIRG1)genes by whole exome sequencing were identified in the patient and her daughter.The missense mutation(c.857G>A)occurred in the CLCN7 gene p.R286Q,which is highly conserved across species.The TCIRG1 gene point mutation(c.714-20G>A)in intron 7(near the splicing site of exon 7)had no effect on subsequent transcription.CONCLUSION This ADO-II case had a pathogenic CLCN7 mutation and late onset without the usual clinical symptoms.For the diagnosis and assessment of the prognosis for osteopetrosis,genetic analysis is advised.
基金Supported by National Science Fundation of China.No.89138970378.
文摘AIMS To clarify whether the lower interferon gamma (IFNγ) production by lymphocytes in patients with liver diseases is due to defects of lymphocytes themselves or of other cofactors such as interleukin-2(IL-2). METHODS Peripheral blood mononuclear cells (PBMCs) from patients with various liver diseases were cultured with or without PHA and IL-2. The cells were harvested and counted and the su- pernatants were tested for IFNγ by a sensitive and quantitative ABC-ELISA. RESULTS IFNγ was not round in serum samples from patients as well as normal individuals. However,in supernatants of non-in- duced and induced PBMCs,IFN7 was detected by ABC-ELISA. In non-induced PBMCs (group 1),the content of IFNγ in super- natants from control,CAH,CPH and HCC was 8.72 μg/L, 5.03 μg/L,6.02 μg/L and 4.91 μg/L respectively. The pro- duction of IFNγ in liver disease was significantly decreased,com- pared to control. In group 2 in which PBMCs were stimulated with PHA,the content of IFNγ was 22.71,17.12,14.54 and 17.63 μg/L respectively. In group 3 in which PBMCs were in- duced by IL-2,the amount of IFN7 in supernatant from control (60.67 μg/L) was much larger than those from CAH (21.70 μg/ L),CPH (24.00 μg/L) and HCC (19.15 μg/L) (P<0.01). Comparing the amount of IFNγ in group 3 (IL-2-induced) with that in group 1 (non-induced),we found that IFNγ production was en- hanced by nearly 4 folds in liver diseases and by over 7 folds in control,Whereas the number of PBMCs,whether from liver dis- eases or from control,was increased by only approximately 3 folds. CONCLUSIONS The decreased production of IFNγ in liver dis- eases including HCC is mainly due to endogenous defects of lym- phocytes though the defects of stimulating cofactors such as IL-2 may also be involved.
文摘AIM To investigate the therapeutic potential of gamma interferon (IFN γ) gene modified human hepatocellular carcinoma (HCC) cells. METHODS The IFN γ gene was introduced retrovirally into four HCC cell lines. Secreted IFN γ activity was assessed using bioassay. The expression of MHC molecules was detected by FACS. Tumorigenicity was analysed by tumor formation in nude mice. RESULTS Four IFN γ gene transduced HCC cell lines secreted different amounts of IFN γ, as in the same case of five clones derived from one HCC cell line. Transduction with IFN γ caused significant increase in the expression of major histocompatibility complex (MHC) antigens on HCC cells. The expression of HLA class Ⅰ was increased by 2-3 times in terms of mean fluorescence intensities, while for class Ⅱ expression, the percentage of positive cells augmented from <10% to >50%. When equal amount of tumor cells were injected into nude mice, the tumorigenicity of some transduced cells decreased dramatically. CONCLUSION IFN γ gene transduction can convert weakly immunogenic HCC cells to activate antitumor immune response, and further pave the way for the future use of such gene modified tumor cells as a modality for the cancer immunotherapy.
基金the National Natural Science Foundation of China, No. 39570249
文摘BACKGROUND： The supernatant of interferon-gamma （IFNγ） co-cultured with neonatal rat cortical glia can promote the cells in embryonic basal forebrain/septal nuclei to differentiate into cholinergic neurons, but the mechanism is still unclear. OBJECTIVE： To analyze the pathways for IFNγ to promote the differentiation of primarily cultured cholinergic neurons in rat embryonic basal forebrain/septal nuclei through culture in different conditioned medium. DESIGN： A controlled experiment taking cells as the observational target. SETTINGS： Department of Biochemistry and Molecular Biology, Youjiang Medical College for Nationalities; Department of Cell Biology, Beijing University Health Science Center. MATERIALS： Sixty-four pregnant Wistar rats for 16 days （250-350 g） and 84 Wistar rats （either male or female, 5-7 g） of 0-1 day after birth were provided by the experimental animal department of Beijing University Health Science Center. Rat IFNγ were provided by Gibco Company; Glial fibrillary acidic protein by Huamei Company. METHODS： The experiments were carried out in the Department of Cell Biology, Beijing University Health Science Center and Daheng Image Company of Chinese Academy of Science from July 1995 to December 2002. ① Interventions： The nerve cells in the basal forebrain/septal nuclei of the pregnant Wistar rats for 16 days were primarily cultured, and then divided into four groups： Blank control group （not any supernatant and medium was added）; Control group （added by mixed glial cell or astrocyte conditioned medium）; IFNγ group （added by mixed glial cell or astrocyte conditioned medium＋IFNγ）. Antibody group （added by mixed glial cell or astrocyte conditioned medium＋IFNγ＋Ab-IFNγ）. Mixed glial cell or astrocyte conditioned medium was prepared using cerebral cortex of Wistar rats of 0-1 day after birth. ② Evaluation： The immunohistochemical method was used to perform the choline acetyltransferase （ChAT） staining of cholinergic neurons. The ChAT positive cells were counted. MAIN OUTCOME MEASURES： Comparison of ChAT positive cells in rat basal forebrain and septal nuclei in different conditioned medium. RESULTS： ① ChAT positive cells in mixed glial cell conditioned medium： The ChAT positive cells in the IFNγ group and antibody group were significantly more than those in the control group （P 〈 0.01）. ② ChAT positive cells in astrocyte conditioned medium： The ChAT positive cells in the IFNγ group were significantly more than those in the control group, but there was no significant difference between the antibody group and control group （P 〉 0.05）. CONCLUSION： IFNγ cannot directly promote the differentiation of cholinergic neurons, but plays a role through activating glial cells （except astrocytes） to produce IFNγ like molecules.
