Epidemiological characteristics of allergic rhinitis and its correlation with serum IFN-γ and IL-4 levels / 公共卫生与预防医学

Objective To analyze the epidemiological characteristics of AR and its correlation with serum IFN-γ and IL-4 levels in Bazhong City, and to provide theoretical basis for clinical prevention and treatment of AR. Methods Adopt the method of multistage stratified random survey from January 2019 to January 2020 bazhong 3 armour hospital otolaryngology seeing a doctor , with face to face questionnaire survey form the people generally, AR number of statistics, the main clinical symptoms and related symptoms, AR merger disease situation, according to the illness severity was divided into mild and moderately severe group, Five mL of elbow venous blood was extracted from AR patients, and 22 kinds of allergens were determined by using the allergen-specific IgE antibody detection kit (western blot). Serum IFN-γ and IL-4 levels were determined by enzyme-linked immunosorbent assay. Pearson correlation analysis was performed on serum IFN-γ and IL-4 levels and severity of allergic rhinitis. Results Among 1 243 patients who completed the questionnaire, the prevalence of AR was 275 (22.12%). There was significant difference in the prevalence of AR among different age groups (χ2=6.809 , P2=7.914 , P<0.0) and the prevalence of AR in workers was the highest (26.48%). Among 275 AR patients in Bazhong City, 153 cases (55.64%) had seasonal allergic rhinitis and 122 cases (44.36%) had perennial allergic rhinitis. The main clinical symptoms of AR patients were sneezing in 234 cases (85.09%) and nasal obstruction in 197 cases (71.64%). AR combined with bronchial asthma in 59 cases (21.45%), conjunctivitis in 28 cases (10.18%); the peak of AR incidence was mainly in July. Among 22 allergens, dust mite was the most common allergen in 139 cases (50.55%), followed by penicillin in 56 cases (20.36%). The main inducing factors were pollen 112 cases (40.73%) and cold 98 cases (35.64%). According to the severity of the disease, they were divided into mild group (n=178) and moderate to severe group (n=97). The serum IL-4 level in moderate and severe groups was significantly higher than that in mild group (P<0.05). The serum IFN-γ level in moderate and severe groups was significantly lower than that in mild group (P<0.05). Pearson correlation analysis showed that il-4 level was positively correlated with disease severity (r=0.492, P<0.05). IFN-γ was negatively correlated with the severity of the disease (r=-0.459, P<0.05). Conclusion The prevalence of AR is high in Bazhong city, and the main clinical symptom is sneezing. Among the complications, bronchial asthma is the most common. Colds and weather changes are the main factors causing AR.

Ficus carica hairy roots: In vitro anti-leishmanial activity against Leishmania major promastigotes and amastigotes

Objective: To investigate the biochemical capacity, and in vitro inhibitory effects of hairy roots from two cultivars of Ficus carica L. (Sabz and Siah) on Leishmania major promastigotes and amastigotes. Methods: In the hairy roots, the activity of antioxidant enzymes compared to normal leaves and roots, and the presence of some phenolic compounds in comparison with fruits were investigated. The IC 50 values of hairy roots in promastigotes was determined by tetrazolium-dye 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and trypan blue assays. By calculating the infectivity index of peripheral blood mononuclear cells (PBMCs), the leishmanicidal activity (IC 50 values) of hairy roots for amastigotes was estimated. The effects of hairy roots (IC 50 values) treatment on the levels of IFN-γ and iNOS expression, intracellular reactive oxygen species, and iNOS protein expression in infected-PBMCs were determined. Results: Based on antioxidant enzyme assays and high performance liquid chromatography analysis, hairy roots exhibited high antioxidant capacity and contained high levels of phenolic compounds. According to the results of tetrazolium-dye 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and trypan blue assays, the hairy root extracts of both cultivars showed considerable dose-dependent inhibitory effects against Leishmania major promastigotes. Depending on the concentration and exposure time, treatment of infected-PBMCs with hairy root extracts caused the generation of a significant reactive oxygen species, up-regulation of IFN-γ and iNOS genes expression, and high value of iNOS protein compared to controls. Conclusions: The findings of this study suggest that the hairy roots of Ficus carica can be considered as a promising natural source of antileishmanial agents.

Screening and identification of H-2 d-restricted T cell epitopes in fusion and attachment glycoproteins of Nipah virus by ELISPOT / 中华微生物学和免疫学杂志

Objective:To screen and identify H-2 d-restricted T cell epitopes in fusion (F) and attachment (G) glycoproteins of Nipah virus (NiV) in mice. Methods:The complete peptides (single peptide contains 15 amino acids, and 10 amino acids were repeated in the front and back peptides) derived from F and G antigens were mixed into peptide libraries. BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination. The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design, and spleen cells of immunized mice were collected and analyzed by IFN-γ ELISPOT assay to detect the dominant H-2 d-restricted epitope peptides. Results:Twelve dominant H-2 d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction. Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction. Conclusions:In this study, 12 F antigen-specific and 4 G antigen-specific H-2 d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γ ELISPOT, which could provide reference for immunological analysis of NiV and vaccine research.

Acupuncture combined with western medication for ocular myasthenia gravis: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical efficacy between acupuncture combined with western medication and simple western medication for ocular myasthenia gravis (OMG), and to explore its possible mechanism.@*METHODS@#A total of 60 patients of ocular myasthenia gravis were randomized into an acupuncture combined with western medication group (30 cases, 1 case dropped off) and a western medication group (30 cases, 2 cases dropped off). Oral pyridostigmine bromide tablet and prednisone acetate tablet were given in the western medication group. On the basis of the treatment in the western medication group, Tongdu Tiaoqi acupuncture (acupuncture for unblocking the governor vessel and regulating qi ) was applied at Baihui (GV 20), Fengfu (GV 16), Hegu (LI 4), Zusanli (ST 36), etc. in the acupuncture combined with western medication group, once a day, 6 days a week. The treatment was given 8 weeks in both groups. Before and after treatment, the OMG clinical absolute score was observed, electrophysiological indexes of orbicularis oculi (value of mean jitter, percentage of jitter >55 μs and percentage of blocks) were measured by single-fiber electromyography (SFEMG), serum levels of acetylcholine receptor antibody (AChR-Ab), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were detected by ELISA method.@*RESULTS@#After treatment, the OMG clinical absolute scores, values of mean jitter, percentages of jitter >55 μs, percentages of blocks and serum levels of AChR-Ab, IFN-γ and IL-4 were decreased compared before treatment in both groups (P<0.05), and those in the acupuncture combined with western medication group were lower than the western medication group (P<0.05).@*CONCLUSION@#Acupuncture combined with western medication can effectively improve ptosis, palpebra superior fatigability, eye movement disorder and neuromuscular junction dysfunction in patients with ocular myasthenia gravis, the therapeutic effect is superior to simple western medication. Its mechanism may be related to down-regulating serum levels of AChR-Ab, IFN-γ and IL-4 and promoting the recovery of orbicularis oculi function.

Effect of Astragali Radix-Curcumae Rhizoma compatibility combined with 5-fluorouracil on Th17/Treg balance and tumor-related mRNA and protein expression in orthotopic xenograft model mice of CT26.WT colorectal carcinoma / 中国中药杂志

Astragali Radix-Curcumae Rhizoma(AR-CR) is a combination commonly used in the clinical treatment of tumors. Based on the T helper 17(Th17)/regulatory T cell(Treg) balance, the present study explored the possible mechanism of AR-CR combined with 5-fluorouracil(5-FU) on the tumor growth of orthotopic xenograft model mice of colorectal carcinoma. Ninety male BALB/c mice were randomly divided into nine groups, i.e., a blank group, a model group, a 5-FU group, high-, medium-, and low-dose AR-CR(2∶1) groups, and high-, medium-, and low-dose AR-CR+5-FU groups, with 10 mice in each group. The orthotopic xenograft model of CT26.WT colorectal carcinoma was induced in mice except those in the blank group. Twenty-four hours after the ope-ration, mice in the blank group and the model group received normal saline by gavage(10 mL·kg~(-1), once per day), and those in the 5-FU group received 5-FU by intraperitoneal injection(25 mg·kg~(-1), once every other day). Mice in the AR-CR groups received AR and CR decoctions by gavage(12, 6, and 3 g·kg~(-1), once a day) and those in the combination groups received AR and CR decoctions and 5-FU(doses and administration methods were the same as above). After intervention for three weeks, all mice were sacrificed and tumor tissues were collected. The tumor mass was weighed and the average tumor weight was calculated. The changing trend of Th17/Treg(%) in the CD4~+T lymphocytes of the spleen tissues of the mice in each group was detected. The mRNA expression in the blood and protein expression in the tumor tissues of transforming growth factor-β(TGF-β), tumor necrosis factor-α(TNF-α), interferon-γ(IFN-γ), Smad4, N-cadherin, matrix metalloproteinase-7(MMP-7) were detected. The experimental results revealed that compared with the model group, the groups with drug intervention showed reduced tumor mass(P<0.01), decreased CD4~+IL-17~+ in the spleen tissues to varying degrees(P<0.001), and increased proportion of CD4~+Foxp3~+(P<0.001 or P<0.05), indicating that Th17/Treg maintained dynamic balance, and the effect of the combination groups was predominant. Additionally, the mRNA expression in the blood and protein expression in the tumor tissues of TGF-β, TNF-α, IFN-γ, Smad4, N-cadherin, and MMP-7 declined to varying degrees in a dose-dependent manner(P<0.01 or P<0.001). The AR-CR combined with 5-FU can inhibit the tumor growth of orthotopic xenograft model mice of CT26.WT colorectal carcinoma. The mechanism may be related to maintenance of Th17/Treg dynamic balance in the body and down-regulation of TGF-β, TNF-α, IFN-γ, Smad4, N-cadherin, and MMP-7 expression.

Effect of American ginseng saponins on enhancing immunity based on zebrafish model organisms / 中草药

Objective: To study the immune enhancing effect of saponins from American ginseng and its mechanism. Methods: The drug dosage of American ginseng saponin extract and the optimal modeling dose of rapamycin were determined by tolerance test and immunodeficiency model establishment experiments. Zebra fish were divided into control group (0.5% dimethyl sulfoxide), model group (4 μg/mL rapamycin), and three origins American ginseng treatment groups with different concentrations (Shandong, Northeast, Canada, 4 μg/mL rapamycin +10, 25 and 50 μg/mL American ginseng saponin extract) 48 h after fertilization (48 hpf). After a certain period of incubation, the number of neutrophils in the tail of zebrafish, macrophage phagocytosis, and interferon gamma (IFN-γ) content in the body were used as indexes, and the growth rate of neutrophils was calculated to investigate the immune-enhancing effect of American ginseng saponins. Results: When the dosage of American ginseng saponins was higher than 50 μg/mL, the mortality of Zbra-fish increased with the increase of the concentration and the time of administration; Compared with control group, in model group, the number of neutrophils in the tail of zebrafish was decreased significantly (P < 0.01), and the content of IFN-γ in the body was decreased significantly (P < 0.01); Compared with model group, in 10 μg/mL and 25 μg/mL American ginseng treatment group, the number of neutrophils in the tail of zebrafish was increased significantly (P < 0.01), the number of macrophages engulfing ink particles was increased significantly (P < 0.01), the content of IFN-γ in vivo was increased significantly (P < 0.01), and the growth rate of neutrophils was between 19.73% and 96.49%. Conclusion: The American ginseng saponins have a better effect on enhancing immunity.

Effect of asbestos exposure on differentiation and function of cytotoxic T lymphocytes

Asbestos exposure is known to cause malignant mesothelioma, which is associated with poor prognosis. We focused on and examined the effect of asbestos exposure on the differentiation and function of cytotoxic T lymphocytes (CTLs). CTLs have the ability to specifically attack tumor cells after being differentiated from naïve CD8 T cells following antigen stimulation. Exposure to chrysotile B asbestos suppressed the differentiation of CTLs during the mixed lymphocyte reaction (MLR) and was associated with a decrease in proliferation of CD8 T cells. Additionally, in an effort to investigate the mechanism associated with suppressed CTL differentiation upon exposure to asbestos, we focused on IL-2, a cytokine involved in T cell proliferation. Our findings indicated that insufficient levels of IL-2 are not the main cause for the suppressed induction of CTLs by asbestos exposure, although they suggest potential improvement in the suppressed CTL function. Furthermore, the functional properties of peripheral blood CD8 lymphocytes from asbestos-exposed individuals with pleural plaque (PP) and patients with malignant mesothelioma (MM) were examined. MM patients showed lower perforin levels in CD8 lymphocytes following stimulation compared with PP-positive individuals. The production capacity of IFN-γ in the MM group tended to be lower compared with healthy volunteers or PP-positive individuals. In an effort to determine whether chronic and direct asbestos exposure affected the function of CD8 T cells, cultured human CD8 T cells were employed as an in vitro model and subjected to long-term exposure to chrysotile (CH) asbestos. This resulted in decreased levels of intracellular perforin and secreted IFN-γ. Those findings underlie the possibility that impaired CD8 lymphocyte function is caused by asbestos exposure, which fail to suppress the development of MM. Our studies therefore reveal novel effects of asbestos exposure on CTLs, which might contribute towards the development and implementation of an effective strategy for the prevention and cure of malignant mesothelioma.

Study on the Effects of Biyuanshu Oral Solution on mRNA Expression of IFN-γ and Immune Checkpoint B7-H1/ PD-1 of Nasal Sinus Mucosa in CRS Model Mice / 中国药房

OBJECTIVE：To investigate the effects of Biyuanshu （BYS）oral solution on IFN-γ of chronic rhinosinusitis （CRS）model mice ，and to investigate its potential mechanism on the basis of B 7-H1/PD-1 immune checkpoint. METHODS ：Male C57 mice were randomly divided into normal group ，sham operation group ，chemical medicine control group （clarithromycin，103 mg/kg），BYS low-dose ，medium-dose and high-dose groups （BYS oral solution ，3.1，6.2，12.4 mL/kg），with 20 mice in each group. Except for normal group without any treatment ，other mice were all open maxillary sinus ，sham operation group was not filled with sponge with bacteria ，while model group and administration groups were filled with sponge with bacteria to induce CRS model. Since 8th week after modeling ，normal group ，sham operation group and model group were given normal saline 0.2 mL intragastrically，administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 14 d. The nasal symptoms and general condition of mice were observed ，and the pathological changes of mice ’s nasal sinus mucosa were observed by HE staining ；qRT-PCR was used to measure the mRNA expression of IFN-γ，B7-H1 and PD- 1 in nasal sinus mucosa of mice. RESULTS：The normal group and sham operation group had no abnormal in nose ，and the epithelium and cilia of the nasal sinus mucosa were intact ；there was no significant difference f8y3j0127@163.com in the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 between 2 groups（P＞0.05）. In model group ，the mice were found to have runny n ose，frequent scratching and sneezing ，a small amount of yellow secretion in the nasal cavity ，and serious depilation ；the nasal sinus mucosa was seriously damaged ，cilia was exfoliated ，and the gland in the submucosa was hyperplasia ，lymphocyte infiltration was also found ；the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 were significantly increased compared with normal group （P＜0.01）. Compared with model group，the nasal symptoms ，general condition and pathological changes of the nasal sinuses in each administration group were improved in varying degrees ；the relative mRNA expression of IFN-γ and B7-H1 in chemical medicine control group ，BYS medium-dose and high-dose groups ，as well as the relative mRNA expression of PD- 1 in administration groups were decreased significantly；above indexes of BYS medium-dose and high-dose groups were significantly lower than BYS low-dose group ，while relative mRNA expression of IFN-γ in BYS high-dose group were significantly higher than BYS medium-dose group. The relative mRNA expression of IFN-γ in BYS low-dose and medium-dose groups，the relative mRNA expression of B 7-H1 in BYS low-dose group，the relative mRNA expression of PD- 1 in BYS groups were significantly higher than chemical medicine control group ； mRNA expression of IFN-γ in BYS high-dose group was significantly higher than chemical medicine control group（P＜0.05 or P＜ 0.01）. Above indexes of BYS medium-dose group were similar to those of chemical medicine control group （P＞0.05）. CONCLUSIONS：BYS oral solution can improve chronic inflammation in nasal sinus mucosa of mice ，the mechanism of which may be associated with intervening mRNA overexpression of B 7-H1/PD-1 by inhibiting mRNA expression of IFN-γ.

Interleukin-27 decreases ghrelin production through signal transducer and activator of transcription 3-mechanistic target of rapamycin signaling

Interleukin-27 (IL-27), a heterodimeric cytokine, plays a protective role in diabetes. Ghrelin, a gastric hormone, provides a hunger signal to the central nervous system to stimulate food intake. The relationship between IL-27 and ghrelin is still unexplored. Here we investigated that signal transducer and activator of transcription 3 (STAT3)-mechanistic target of rapamycin (mTOR) signaling mediates the suppression of ghrelin induced by IL-27. Co-localization of interleukin 27 receptor subunit alpha (WSX-1) and ghrelin was observed in mouse and human gastric mucosa. Intracerebroventricular injection of IL-27 markedly suppressed ghrelin synthesis and secretion while stimulating STAT3-mTOR signaling in both C57BL/6J mice and high-fat diet-induced-obese mice. IL-27 inhibited the production of ghrelin in mHypoE-N42 cells. Inhibition of mTOR activity induced by siRNA or rapamycin blocked the suppression of ghrelin production induced by IL-27 in mHypoE-N42 cells. siRNA also abolished the inhibitory effect of IL-27 on ghrelin. IL-27 increased the interaction between STAT3 and mTOR in mHypoE-N42 cells. In conclusion, IL-27 suppresses ghrelin production through the STAT3-mTOR dependent mechanism.

The effect of Ganciclovir Pellicles in patients with viral keratitis and the effect on serum levels of inflammatory factors / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To study the effect of biological keratoconjunctivitis combined with Ganciclovir Pellicles in patients with infectious keratitis and the effect on serum levels of inflammatory factors.<p>METHODS: In this study, 54 patients with 54 eyes were selected as the research object and divided into the observation group and the control group according to the random number table method. All patients were treated with bioartificial corneal transplantation. The patients in the observation group were treated with Ganciclovir eye drops, and the control group was treated with Aciclovir eye drops. The changes of LogMAR visual acuity, corneal transparency, serum inflammatory factors and cytokines were analyzed.<p>RESULTS: The rates of getting rid of blindness in the observation group and the study group were 93% and 89% respectively(<i>P</i>=0.642). The LogMAR visual acuity of two groups were significantly improved in 1wk and 6mo after operation compared with that before operation(<i>P</i><0.05). At 7d after operation, the levels of IL-6, TNF-α, INF- γ, SOD, NO and MDA in the two groups were significantly improved, and the improvement of the indexes in the observation group was better than that of the control group(<i>P</i><0.05). The rate of adverse reaction rate between two groups showed no significant difference(<i>P</i>=0.556).<p>CONCLUSION: Biological keratoconus combined with Ganciclovir has a good effect on viral keratitis, which can improve vision, inflammation and reduce the level of MDA.

Interferon-γ regulates cell malignant growth via the c-Abl/HDAC2 signaling pathway in mammary epithelial cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Interferon-γ (IFN-γ) has been used to control cancers in clinical treatment. However, an increasing number of reports have suggested that in some cases effectiveness declines after a long treatment period, the reason being unclear. We have reported previously that long-term IFN-γ treatment induces malignant transformation of healthy lactating bovine mammary epithelial cells (BMECs) in vitro. In this study, we investigated the mechanisms underlying the malignant proliferation of BMECs under IFN-γ treatment. The primary BMECs used in this study were stimulated by IFN-γ (10 ng/mL) for a long term to promote malignancy. We observed that IFN-γ could promote malignant cell proliferation, increase the expression of cyclin D1/cyclin-dependent kinase 4 (CDK4), decrease the expression of p21, and upregulate the expression of cellular-abelsongene (c-Abl) and histone deacetylase 2 (HDAC2). The HDAC2 inhibitor, valproate (VPA) and the c-Abl inhibitor, imatinib, lowered the expression level of cyclin D1/CDK4, and increased the expression level of p21, leading to an inhibitory effect on IFN-γ-induced malignant cell growth. When c-Abl was downregulated, the HDAC2 level was also decreased by promoted proteasome degradation. These data suggest that IFN-γ promotes the growth of malignant BMECs through the c-Abl/HDAC2 signaling pathway. Our findings suggest that long-term application of IFN-γ may be closely associated with the promotion of cell growth and even the carcinogenesis of breast cancer.

Study on Improvement Effects of Qishenlian Eczema Cream on the Atopic Dermatitis Model Guinea Pigs / 中国药房

OBJECTIVE： To investigate the improvement effects of Qishenlian eczema cream on the atopic dermatitis （AD） model guinea pigs， and to investigate its possible mechanism. METHODS： Female guinea pigs were randomly divided into normal control group， model control group， matrix control group （Qishenlian eczema cream matrix， 1 g/kg）， Qishenlian eczema cream low-dose， medium-dose and high-dose groups （0.5， 1， 2 g/kg） and Hydrocortisone butyrate cream group （0.5 g/kg）， with 10 guinea pigs in each group. Using ovalbumin as antigen， the skin allergy of guinea pigs was stimulated to induce AD model. Forty-eight hours after the last excitation， guinea pigs were smeared with normal saline in normal control group and model control group， while those were smeared with Qishenlian eczema cream matrix in matrix control group， and administration groups were treated with relevant medicine， twice a day， for consecutive 14 d. The scores of erythema， edema and scratch before and after administration were recorded in each group. The morphological characteristics of skin were observed by HE staining. TUNEL method was used to detect the apoptosis of keratinocytes in skin tissue. The level of IFN-γ in skin lesions was detected by ELISA. RESULTS： Compared with normal control group， inflammatory cell infiltration， hyperkeratosis of epidermis and thickening of spinous layer were observed in skin tissue of guinea pigs in model control group and matrix control group； the scores of erythema， edema and scratch and their total score， the level of IFN-γ in skin tissue were increased significantly； the apoptosis number of keratinocytes was decreased significantly （P＜0.01）. Compared with model control group and matrix control group， above symptoms of medication groups were relieved to different extents； the scores of erythema， edema and scratch and their total score after medication in medication groups， the level of IFN-γ in skin tissue in Qishenlian eczema cream medium-dose and high-dose groups and Hydrocortisone butyrate cream group were decreased significantly， and above scores in medication groups were significantly lower than before medication （P＜0.05 or P＜0.01）； the apoptosis number of keratinocytes was increased significantly in medication groups （P＜0.05 or P＜0.01）. CONCLUSIONS： Qishenlian eczema cream has certain improvement effect on AD model guinea pigs， the mechanism of which may be associated with promoting the keratinocyte apoptosis， reducing the level of INF-γ in skin tissue.

Anti-tumor experimental study of mouse multi-subtype heat shock protein/peptide vaccine combined with PD-L1 immunological checkpoint inhibitor / 中国肿瘤临床

Objective: To evaluate the anti-tumor activity of mouse multi-subtype heat shock protein/peptide (mHSP/P) vaccine in combination with a programmed death ligand 1 (PD-L1) inhibitor in mouse sarcoma. Methods: Immunohistochemical staining and en-zyme-linked immunosorbent assay (Elisa) was used to quantitatively identify the expression of heat shock proteins (HSP70, HSP90, Grp94) in the sarcoma cell line MCA207. From the protein suspension prepared, mHSP/P and Grp94/peptide (Grp94/P) sarcoma vac-cines were isolated using chromatography and were identified by Western blot (WB). Flow cytometry was used to determine their cy-totoxic effects. The levels of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) produced upon mHSP/P and Grp94/P stimulation were measured by Elisa. The effect of sarcoma vaccines on the growth and survival of sarcoma was evaluated in mice. The expression of PD-L1 on the surface of MCA207 sarcoma cells was evaluated by immunofluorescent staining. The effect of IFN-γ treatment on the expression of PD-L1 was determined by WB. Animal experiments explored the effects of PD-L1 inhibitor in combination with mHSP/P treatment on tumors. Results: Tumor tissue carries a variety of HSP subtypes (HSP70, HSP90, Grp94). We successfully isolated sarco-ma tissue-derived mHSP/P and Grp94/P tumor vaccines, which were identified by WB; flow cytometry analysis demonstrated their cy-totoxicity. The levels of IFN-γ and TNF-α cytokines upon mHSP/P stimulation were significantly higher than that observed upon Grp94/P stimulation (P<0.05). The expression of PD-L1 on the surface of sarcoma cells increased with IFN-γ treatment. Animal experiments demonstrated that PD-L1 inhibitor in combination with mHSP/P significantly increased the immune response against tumor (P<0.05). Conclusions: Tumor-derived mHSP/P and Grp94/P can be used as tumor vaccines in animal models. The mHSP/P can elicit a stronger anti-tumor immune response than Grp94/P. IFN-γ stimulates the expression of PD-L1 in sarcoma cells, which results in immune eva-sion. The PD-L1 inhibitor in combination with mHSP/P increased the anti-tumor effect in the tumor microenvironment.

Effect of acupuncture on the expressions of TNF-α and IFN-γ in patients with premature ovarian failure / 中国针灸

OBJECTIVE@#To explore the clinical effect of acupuncture and the potential effect mechanism in patients with premature ovarian failure.@*METHODS@#A total of 104 patients with premature ovarian failure were randomized into an acupuncture group and a western medication group, 52 cases in each one. In the western medication group, the conjugated estrogens tablets were prescribed for oral administration, 0.625 mg each time, once a day, consecutively for 21 days. On the 16th day of medication with conjugated estrogens tablets, the oral administration of medroxyprogesterone acetate tablets were supplemented, 10 mg each time, once a day, consecutively for 5 days, and then, these two kinds of western medication were discontinued for 1 week. A total of 3 cycles were required in treatment with 28 days as an artificial cycle. In the acupuncture group, acupuncture was applied. Two groups of acupoints were selected. The first group of acupoints were stimulated before ovulation and the acupoints were Guanyuan (CV 4), Guilai (ST 29), Taichong (LR 3), Taixi (KI 3), Xuehai (SP 10), Sanyinjiao (SP 6), Zigong (EX-CA 1), Yinlingquan (SP 9), Zusanli (ST 36), Shuidao (ST 28), Dahe (KI 12) and Tianshu (ST 25). The second group of acupoints were stimulated after ovulation and the acupoints included Ciliao (BL 32), Shiqizhui (EX-B 8), Ganshu (BL 18), Shenshu (BL 23), Geshu (BL 17) and Pishu (BL 20). The therapeutic effect was observed and compared in the patients between the two groups, as well as the expressions of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) and the levels of serum luteinizing hormone (LH), follicule stimulating hormone (FSH) and estradiol (E) before and after treatment.@*RESULTS@#The total effective rate was 90.4% (47/52) in the acupuncture group, higher than 67.3% (35/62) in the western medication group (<0.05). After treatment, the expressions of IFN-γ and TNF-α in the acupuncture group were obviously lower than the western medication group (<0.05). Except for serum LH after treatment, at the end of treatment and in 30 days and 90 days after treatment, the levels of serum E in the acupuncture group were higher obviously than the western medication group and the levels of serum LH and FSH were lower obviously than the western medication group (all <0.05).@*CONCLUSION@#Acupuncture promotes the regular menstruation, effectively regulates the levels of serum LH, FSH and E and improves the pituitary gland and the ovary endocrine in the patients with premature ovarian failure. Such effect may be related to the the improvements in the expressions of IFN-γ and TNF-α, the inhibition of the apoptosis of ovarian granulosa cells, the recovery of ovarian function and the enhancement of reserve capacity.

IL-17A involved in respiratory syncytial virus-associated pathogenesis by promoting IFN-γand inhib-iting viral clearance in mice / 中华微生物学和免疫学杂志

Objective To identify the role of IL-17A during respiratory syncytial virus (RSV) in-fection in a mouse model. Methods Female wild-type C57BL/6 mice and IL-17A knockout ( IL-17A-/-) mice at the age of 6 to 8 weeks were both randomly divided into two groups:control and RSV groups. Mice in the control groups were given the supernatant of Hep-2 cell culture, while those in the RSV groups were treated with RSV A2 through intranasal administration. Leukocytes in bronchoalveolar lavage fluid ( BALF) samples were counted. Left lung tissues were stained with hematoxylin and eosin ( HE ) to evaluate his-topathological scores. Airway hyperresponsiveness ( AHR) was measured by whole-body plethysmography. The concentrations of IFN-γ were determined with ELISA. RSV titers were measured by plaque assay. To assess the effects of IL-17A on IFN-γproduction and its role in RSV infection, IL-17A-/- mice were treated with exogenous recombinant murine IFN-γ or IL-17A, while wild-type mice were given IFN-γ neutralizing antibody intervention. Results The counts of inflammatory cells and neutrophils in BALF, lung tissue his-topathological scores, AHR, IFN-γlevels and virus titers of the wild-type group were higher than those of the IL-17A-/-group after RSV infection. IFN-γlevels, inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly increased in RSV-infected IL-17A-/- mice after the intervention of recombinant murine IL-17A or IFN-γ. RSV titers were much higher in the recombinant murine IL-17A-trea-ted group, but not affected by the recombinant murine IFN-γ intervention. Inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly decreased in RSV-infected wild-type mice following IFN-γ neutralizing antibody treatment, but no significant changes were found in RSV titers. Conclusions IL-17A might be involved in the pathogenesis of pulmonary diseases during RSV infection through promoting IFN-γ production and inhibiting viral clearance in mice.

Expression of serum Th1/Th2 cytokines in different stages of hepatitis B and its clinical significance / 国际检验医学杂志

Objective To investigate the expression levels of IFN-γ, IL-4, IL-6 in the plasma of HBV chronic patients.To acknowledge the correlation between the immune function of T-cell with chronic hepatitis B virus.Methods The expression levels of IFN-γ, IL-4, IL-6 in the plasma were determined by ELISA in 42 cases of chronic hepatitis B (CHB group), 30 cases of cirrhosis (LC group), 30 cases of primary liver cancer (HCC group) and 30 cases of health control (NC group).Combine with the load of HBV DNA, HBeAg and ALT, the expression levels of IFN-γ, IL-4, IL-6 were re-analysed.Results The expression levels of IFN-γ, IL-4 and IL-6 in the researched groups were statistically significantly different.The expression levels of IFN-γ, IL-4, IL-6 in CHB, LC and HCC groups were higher than NC group with significance.The expression level of IL-4 in the CHB group was higher than LC and HCC groups with significance.The expression level of IL-6 in the HCC group was higher than CHB and LC groups with significance.Compared with the control group, the ratio of IFN-γ/IL-4 in CHB, LC and HCC groups were decreased with significance.There was significantly positive correlation between the expression level of IL-4 with the load of HBV DNA (r=0.757, P＜0.001).The expression level of IFN-γhad negative correlation between the load of HBV DNA (r=-0.786, P＜0.001) with the level of ALT (r=-0.713, P＜0.001) with significance.There was no significant correlation between HBeAg with the cytokines in the serum of patients with chronic HBV.Conclusion The results showed the immune function of T-cell play important role in chronic HBV patients.That would provide a good basis on CHB treatment.The expression levels of IL-6 were higher in HCC group with significance, which provided the direction for the deep study of liver cancer.

A Feasibility Study for Diagnosis of Latent Tuberculosis Infection Using an IGRA Point-of-Care Platform in South Korea

PURPOSE: This study aimed to evaluate ichroma™ IGRA-TB, a novel point-of-care platform for assaying IFN-γ release, and to compare it with QuantiFERON-TB Gold In-Tube (QFT-GIT) for identifying Mycobacterium tuberculosis (M. tb) infection. MATERIALS AND METHODS: We recruited 60 healthy subjects, and blood samples were obtained in QFT-GIT blood collection tubes. The blood collection tubes were incubated at 37℃, and culture supernatant was harvested after 18–24 hours. IFN-γ responses were assessed by the ichroma™ IGRA-TB cartridge and the QFT-GIT IFN-γ enzyme-linked immunosorbent assay. Three active TB patients were recruited as a positive control for M. tb infection. RESULTS: The area under the receiver operating characteristic curve of the ichroma™ IGRA-TB test for differentiating between infected and non-infected individuals was 0.9706 (p < 0.001). Inconsistent positivity between the two tests was found in three participants who showed weak positive IFN-γ responses ( < 1.0 IU/mL) with QFT-GIT. However, the two tests had excellent agreement (95.2%, κ=0.91, p < 0.001), and a very strong positive correlation was observed between the IFN-γ values of both tests (r=0.91, p < 0.001). CONCLUSION: The diagnostic accuracy demonstrated in this study indicates that the ichroma™ IGRA-TB test could be used as a rapid diagnostic method for detecting latent TB infection. It may be particularly beneficial in resource-limited places that require cost-effective laboratory diagnostics.

Therapeutic Efficacy of Mesenchymal Stem Cells and Mesenchymal Stem Cells-derived Neural Progenitors in Experimental Autoimmune Encephalomyelitis

BACKGROUND AND OBJECTIVES: The goal of treatment for MS is to reduce the inflammation and induce the regeneration of degenerated axons. Considering the anti-inflammatory and regenerative capacity of mesenchymal stem cell (MSCs), in this study the therapeutic efficacy of allogeneic MSCs and MSCs-derived neural progenitor cells (MSCs-NPs) was investigated in cellular therapy of chronic experimental autoimmune encephalomyelitis (EAE). METHODS AND RESULTS: MSCs, MSCs-NPs and MSCs+MSCs-NP were administered intravenously to EAE mice on days 22, 29, and 36 post immunization. The levels of cytokines and PGE2 in sera or supernatant of in vitro cultured splenocytes derived from treated mice were measured by ELISA. The results of this study showed that in comparison to MSCs monotherapy, MSCs-NPs administration had a more profound capability of inhibiting the proliferation of pathogenic MOG35–55-specific T cells, decreasing IFN-γ production and increasing anti-inflammatory IL-10 cytokine production. These findings could be explained by higher ability of in vitro cultured MSCs-NPs in production of PGE2 compared to MSCs. In line with these findings, while the administration of MSCs and MSCs-NPs significantly decreased the clinical scores of EAE in comparison with the untreated EAE group, MSCs-NPs were significantly more efficient in reducing clinical score compared to MSCs. Of interest, combined therapy with MSCs and MSCs-NPs did not provide any benefit over monotherapy with MSCs-NPs. CONCLUSIONS: In comparison to MSCs, allogenic MSCs-NPs are more potent in the attenuation of EAE.

Intracellular cytokine staining (ICS) for detection of NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) / 中华微生物学和免疫学杂志

Objective To analyze the possibility of using intracellular cytokine staining (ICS) to evaluate NK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) and to detect the changes in ADCC activity among patients with chronic HIV and/or HCV infection. Methods Flow cytometry was per-formed to determine the percentages of NK cells and the expression of NK cell receptors. ImageStreamX MarkⅡ system was used to identify the expression of CD3, CD56, CD16 and CD32 on CD56brightNK and CD56dimNK subsets. Degranulation process and cytokine production in NK cells were detected using an anti-gen-antibody complex model of P815/Ab in combination with ICS. Differences in NK cell-mediated ADCC were evaluated among patients infected with HIV and/or HIV and healthy subjects by flow cytometry. Re-sults The percentages of CD107a+and IFN-γ+NK cells were positively correlated with the decrease of mean fluorescence intensity (MFI) of CD16. ICS assay revealed a positive correlation between the secretion of CD107a and IFN-γ by NK cells. CD16 was highly expressed in CD56dimNK cells. The ADCC mediated by CD56dimNK cells was stronger than that mediated by CD56brightNK cells. The rate of target cell lysis detected by rapid fluorescence assay was positively correlated with the percentage of CD107a+/IFN-γ+NK cells meas-ured by ICS. NK cell-mediated ADCC was suppressed in patients with chronic HIV and/or HCV infection. Conclusion This study suggests that ICS assay could be used to evaluate NK cell-mediated ADCC. It also reveals that NK cell-mediated ADCC is suppressed in patients with chronic HIV and/or HCV infection.

Effects of Jianpi Qingchang Decoction Combined with Calcitriol on IL-17 and IFN-γ in Ulcerative Colitis Rats / 中国中医药信息杂志

Objective To investigate the effects of Jianpi Qingchang Decoction combined with Calcitriol on the expressions of IL-17 and IFN-γ in colitis in rats with ulcerative colitis; To discuss the mechanism of action. Methods Experimental rats were randomly divided into control group, model group, Mesalazine group, Calcitriol group, Jianpi Qingchang Decoction+Calcitriol group, with 10 rats in each group. Except for the control group, other groups were given TNBs for modeling. The administration began on the 3rd day after modeling. All rats were weighed and sacrificed on the 11th day after administration. The disease activity index (DAI) were calculated and the colon histopathological (HPS) were evaluated under microscope. The mRNA and protein expressions of IFN-γ and IL-17 in colonic mucosa were detected by RT-PCR and immunohistochemistry. Results Compared with the control group, the DAI and HPS of the model group increased significantly (P<0.01); Compared with the model group, the DAI and HPS of administration groups were reduced significantly (P<0.05, P<0.01), and the mRNA and protein expressions of IFN-γ and IL-17 in Mesalazine group and Jianpi Qingchang Decoction+Calcitriol group decreased significantly (P<0.01). Conclusion Jianpi Qingchang Decoction combined with Calcitriol can down-regulate IL-17 expression, decrease the activity of immune cells, reduce the secretion of inflammatory cells, and lessen the damage of colonic mucosa by regulating Th17 cells.