[Effect of ectodysplasin-A1 on proliferation and cell cycle of ameloblast-like cell].

Objective: To investigate the effects of ectodysplasin-A1 (EDA1) on the proliferation and cell cycle of ameloblast-like epithelial cells (LS8 cells). Methods: Wild EDA1 plasmid pCR3-Flag-EDA1-W (wild group), syndrome mutant EDA1 plasmid pCR3-Flag-EDA1-H252L (mutant group) and empty vector plasmid pCR3-Flag (control group) were transfected into LS8 cells. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry. All tests were repeated three times. Results: Compared with the control group (0.105±0.032), the proliferation activity of the wild group (0.201±0.009) was significantly higher after 72 h (P<0.05). Compared with the control group (0.168±0.054) and the mutant group (0.194±0.059), the proliferation activity of the wild group (0.386±0.066) was significantly higher after 96 h (P<0.05). There was no significant difference between the mutant group and the control group at all time points (P>0.05). In the G0/G1 phase, compared with the control group (65.4%±2.1%) and the mutant group (66.6%±3.1%), the cell distribution ratio of the wild group (51.2%±1.1%) was significantly lower (P<0.01). In the S phase, compared with the control group (23.1%±2.0%) and the mutant group (21.9%±1.8%), the cell distribution ratio of the wild type group (37.3%±2.4%) was significantly higher (P<0.01). There was no significant difference in cell cycle distribution between the mutant group and the control group (P<0.05). Conclusions: Wild EDA1 promotes the proliferation of LS8 cells and the transformation from G0/G1 to S phase. The syndrome mutant EDA1 (EDA1-H252L) loses its function of regulating the cell proliferation and cell cycle of LS8 cells.

[Correlation between Toxoplasma gondii infection and spontaneous abortion in pregnant women: a case-control study].

OBJECTIVE: To examine the correlation between Toxoplasma gondii infection and spontaneous abortion among pregnant women, so as to provide the evidence for the developmentofpreventivemeasuresforspontaneousabortion. METHODS: A total of 228 serum samples collected from women with spontaneous abortion for the first time from January 2018 to December 2019 were selected as the case group, while 228 serum samples collected from pregnant women with a normal delivery and without a history of abortion during the same period were selected as the control group. The serum IgG and IgM antibodies against T. gondii were detected and compared in both groups, and the correlation between T. gondii infection and spontaneous abortion was evaluated. RESULTS: There were no significant differences between the case and control groups in terms of age, education levels, occupation, residency and proportion of keeping cats (all P values > 0.05). The positive rate of anti-T. gondii IgM antibody was significantly higher in the case group than in the control group (adjusted χ2 = 4.08, P < 0.05; OR = 8.25), while no significant difference was seen between the case and control groups (χ2 = 0.42, P > 0.05). CONCLUSIONS: Acute maternal T. gondii infection may remarkably increase the chance of spontaneous abortion. Progestational health education regarding toxoplasmosis prevention and control knowledge and detection of T. gondii infection during pregnancy should be strengthened.

[Analysis of clinical diagnosis and treatment in chronic hepatitis B combined with autoimmune hepatitis].

Objective: To summarize the clinical diagnosis and therapeutic method in chronic hepatitis B (CHB) combined with autoimmune hepatitis (AIH). Methods: Clinical manifestations, laboratory examination, imaging, histopathological characteristics, treatment and prognosis of 19 cases diagnosed with CHB combined with AIH followed at the outpatient Department of Gastroenterology of Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine during December 2013 to June 2018 were retrospectively analyzed. Paired sample t-test was used before and after treatment for the measurement of normal distribution data. Measurement data of non-normal distribution were expressed as a median (quartile spacing) and Wilcoxon matched-pairs signed rank test was used before and after treatment. Results: Among the 19 cases, 5 were male and 14 were female. The age of onset was 35 to 63 years, and the average age was 47.10 ± 8.76 years. There were 12 cases diagnosed with CHB before AIH, 5 cases diagnosed with AIH before CHB, and 2 cases diagnosed with AIH and CHB at the same time. After the definite diagnosis of CHB combined with AIH, nucleoside (acid) analogues (antiviral against hepatitis B virus) combined glucocorticoid therapy were given, and azathioprine or mycophenolate mofetil (immunosuppressant) was added according to the intrahepatic inflammation (inflammation graded at G3 and above) and leukocyte conditions. The duration of treatment varied between 2 weeks to 16 (median treatment duration of 6 weeks), except for one case who was just diagnosed and followed up. Biochemical indicators and immunoglobulin of the remaining 18 cases before and after treatment was significantly decreased, and the differences were statistically significant (P < 0.05), with HBV DNA < 20 copies/ml. Conclusion: CHB combined with AIH diagnosis can be easily missed. Therefore, it requires comprehensive diagnosis combined with clinical characteristics, autoantibodies, and immunoglobulin levels with special emphasis on pathological characteristics of liver tissue. Anti-HBc-positive patients using immunosuppressant should be carefully monitored for HBV DNA and anti-HBV treatment should be given if necessary.

The immune evasion strategies of fish viruses.

Viral infection of a host rapidly triggers intracellular signaling events that induce interferon production and a cellular antiviral state. Viral diseases are important concerns in fish aquaculture. The major mechanisms of the fish antiviral immune response are suggested to be similar to those of mammals, although the specific details of the process require further studies. Throughout the process of pathogen-host coevolution, fish viruses have developed a battery of distinct strategies to overcome the biochemical and immunological defenses of the host. Such strategies include signaling interference, effector modulation, and manipulation of host apoptosis. This review provide an overview of the different mechanisms that fish viruses use to evade host immune responses. The basic mechanisms of immune evasion of fish virus are discussed, and some examples are provided to illustrate particular points.

Chemerin affects the metabolic and proliferative capabilities of chondrocytes by increasing the phosphorylation of AKT/ERK.

OBJECTIVE: The purpose of the present study was to explore the mechanism of action of the adipokine chemerin in osteoarthritis (OA) by means of an in vitro OA model. MATERIALS AND METHODS: Primary chondrocytes were isolated from normal rats. The chondrocytes were stimulated with interleukin 1 beta (IL-1ß, 10 µg/L) to establish a model of induced OA. Chemerin was administered to cells of this model. After culture of the chondrocytes in the presence of chemerin for 48 h, the expression of the genes related to OA occurrence and protection, matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-3 (MMP-3), and matrix metalloproteinase-13 (MMP-13) was examined. Western blot was then performed to analyze the phosphorylation of the AKT and extracellular signal-regulated kinase (ERK) proteins in chondrocytes. RESULTS: Stimulation of chondrocytes with IL-1ß markedly reduced the proliferative capability of chondrocytes. Chemerin (5 µM) also significantly decreased the proliferative capability of chondrocytes. The combined administration of IL-1ß and chemerin induced an even greater reduction in the proliferative capability of chondrocytes. Polymerase chain reaction (PCR) results showed that both IL-1ß and chemerin reduced the expression of the protective genes in OA (MMP-1, MMP-3, and MMP-13). Also, the stimulation with IL-1ß and chemerin significantly enhanced the phosphorylation of AKT/ERK in chondrocytes. CONCLUSIONS: This adipokine induces changes in the metabolic and proliferative capabilities of chondrocytes by increasing the phosphorylation of AKT/ERK, thereby inducing OA or aggravating the symptoms of OA.

[Coronary microvascular resistance and its relevant factors in patients with moderate coronary stenosis and chest pain].

Objective: To evaluate the impact of cardiovascular risk factors on index of microvascular resistance (IMR) and coronary flow reserve (CFR) and to explore the characteristics of IMR and CFR and the relationship between IMR and angiographic features in patients with intermediate coronary stenosis and chest pain. Methods: Fractional flow reserve (FFR), CFR, and IMR were measured in patients who underwent invasive coronary angiography with 40%-70% stenosis by visual assessment. All patients with FFR>0.75 were enrolled and grouped with the cut-off points of IMR≥25 and CFR≤2.0. Patients with IMR≥25 were group H, including two sub-groups (high IMR-low CFR, group H1 and high IMR-high CFR, group H2), while those with IMR<25 were group N. The thrombolysis in myocardial infarction (TIMI) frame were counted. Results: A total of 34 patients with FFR>0.75 were enrolled with 61.8%(21 cases) of males and 38.2% (13 cases) of females. The mean age was (57.3±8.1) years old. High IMR accounted for 47.1% of all cases. There was significant difference between group H and N in TIMI frame (33.0 vs. 20.8, P=0.031). There were significant differences between group H1 and H2 in homocysteine (17.8 µmol/L vs. 12.0 µmol/L, P=0.005) and IMRcorr (58.0 vs. 36.1, P=0.002). IMRcorr was correlated to TIMI frame (r=0.40, P=0.012) for all cases. The sensitivity and specificity of inferring IMR≥35.3 by TIMI frame were 0.75 and 0.65 (P=0.049) with TIMI frame over 40.5. Conclusions: High IMR may be one of the reasons for chest pain in patients with intermediate coronary stenosis. There is no correlation between vascular risk factors and IMR or CFR, while there is positive correlation between TIMI frame and IMR. The specificity is 65% for inferring IMR rise with TIMI frame over 40.5.

The effect of curcumin on NF-κB expression in rat with lumbar intervertebral disc degeneration.

OBJECTIVE: To observe the effect of curcumin on the expression levels of nuclear factor κB-p65 (NF-κB-p65) and tumour necrosis factor α (TNF-α) in the nucleus pulposus in rats with lumbar intervertebral disc degeneration. And to investigate of the mechanism underlying the role of curcumin in decelerating the process of lumbar intervertebral disc degeneration. MATERIALS AND METHODS: The model of lumbar intervertebral disc degeneration was established in Sprague-Dawley (SD) rats followed by a curcumin treatment. The ultra-microstructure histomorphological variations in the lumbar intervertebral disc of SD rats were evaluated. The protein and gene expression levels of NF-κB-p65 and TNF-α in the lumbar intervertebral disc were measured. RESULTS: Magnetic resonance imaging (MRI) and histomorphology confirmed the establishment of a successful lumbar intervertebral disc degeneration model. The results from the MRI and the ultra-microstructures revealed a significant improvement in lumbar intervertebral disc degeneration in the curcumin-treated groups (low dose and high dose). No significant change was observed in the solvent control group treated with dimethyl sulfoxide (DMSO) alone. Based on the results of Western blot analysis and real-time PCR, the curcumin treatment (low dose and high dose) significantly reduced the expression levels of NF-κB-p65 and TNF-α in the lumbar intervertebral disc tissue compared with the groups without curcumin treatment and with the DMSO treatment alone. No significant difference, however, was observed between the low-dose and high-dose curcumin treatment groups. CONCLUSIONS: Curcumin may inhibit the activation of NF-κB by inhibiting the translocation of NF-κB-p65 and reducing the release of inflammatory factors which, thereby, decelerates the process of lumbar intervertebral disc degeneration.

Riboflavin supplementation improves energy metabolism in mice exposed to acute hypoxia.

This study investigated the effects of riboflavin on energy metabolism in hypoxic mice. Kunming mice were fed diets containing riboflavin at doses of 6, 12, 24 and 48 mg/kg, respectively for 2 weeks before exposure to a simulated altitude of 6000 m for 8 h. Changes of riboflavin status and energy metabolism were assessed biochemically. Simultaneously, a (1)H nuclear magnetic resonance (NMR) based metabolomic technique was used to track the changes of plasma metabolic profiling. It was found that the content of hepatic riboflavin was decreased and erythrocyte glutathione activation coefficient was elevated significantly under hypoxic condition. Meanwhile, increased plasma pyruvate, lactate, beta-hydroxybutyrate and urea, as well as decreased plasma carnitine were observed. Riboflavin supplementation improved riboflavin status remarkably in hypoxic mice and decreased plasma levels of pyruvate, free fatty acids and beta-hydroxybutyrate significantly. Plasma carnitine was increased in response to riboflavin supplementation. Results obtained from (1)H NMR analysis were basically in line with the data from biochemical assays and remarkable changes in plasma taurine, choline and some other metabolites were also indicated. It was concluded that riboflavin requirement was increased under acute hypoxic condition and riboflavin supplementation was effective in improving energy metabolism in hypoxic mice.

Proteomic analysis of human bone marrow mesenchymal stem cells transduced with human telomerase reverse transcriptase gene during proliferation.

OBJECTIVES: Previous studies have reported immortalization and tumorigenicity of human mesenchymal stem cells (hMSCs) transduced with exogenous human telomerase reverse transcriptase (hTERT). We also have established a line of hMSCs transduced with hTERT (hTERT-hMSCs) and we have cultured these cells for 290 population doublings (PDs) during which they demonstrated a large proliferation potential but with no tumorigenicity. The aim of this study was to investigate the protein expression profile of hTERT-hMSCs with two-dimensional gel electrophoresis and peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to be able to analyse the effects of exogenous hTERT on protein expression in hMSCs. MATERIALS AND METHODS: We generated proteome maps of primary hMSCs and hTERT-hMSCs at PD 95 and PD 275. RESULTS: A total of 1543 +/- 145 protein spots in gels of primary MSCs at PD 12, 1611 +/- 186 protein spots in gels of hTERT-hMSCs at PD 95 and 1451 +/- 126 protein spots in gels of hTERT-hMSCs at 275 PD were detected. One hundred of these were successfully identified, including 20 which were differentially expressed. CONCLUSIONS: The results suggest that sustaining levels of prohibitin and p53 expression along with differential expression of proteins in hTERT-hMSCs provide an insight into lack of transforming activity of hTERT-hMSCs during cell proliferation.

HIV-1 infection of placental cord blood monocyte-derived dendritic cells.

Dendritic cells (DC), the most potent antigen-presenting cells (APC), have been implicated as the initial targets of HIV infection in skin and mucosal surfaces. DC can be generated in vitro from blood-isolated CD14(+) monocytes or CD34(+) hematopoietic progenitor cells in the presence of various cytokines. In this study, we investigated whether monocytes obtained from placental cord blood are capable of differentiation into dendritic cells when cultured with a combination of cytokines - granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), and tumor necrosis factor-alpha (TNF-alpha). We then examined HIV infection, HIV receptor (CD4, CCR5) expression, and beta-chemokine [macrophage inflammatory protein-1alpha and -1beta (MIP-1alpha, MIP-1beta)] production by placental cord monocyte-derived dendritic cells (MDDC) as compared to that of autologous cord monocyte-derived macrophages (MDM). Monocytes isolated from placental cord blood differentiate into DC after 7 days in culture with the mixture of cytokines, as demonstrated by development of characteristic DC morphology, loss of CD14 expression, and gain of CD83, a marker for mature DC. Mature cord MDDC had significantly lower susceptibility to M-tropic ADA (CCR5-dependent) envelope-pseudotyped HIV infection in comparison to autologous placental cord MDM, whereas there was no significant difference in virus replication in cord MDDC and MDM infected with murine leukemia virus envelope-pseudotyped HIV (HIV receptor-independent). This limited susceptibility of cord MDDC to M-tropic HIV infection may be due to lower expression of CD4 and CCR5 on the cell membrane and higher production of MIP-1alpha and MIP-1beta. These data provide important information toward our understanding of the biological properties of cord MDDC in relation to HIV infection.

[Effects of zinc on the free radical signals and apoptosis in liver with ischemia/ reperfusion injury].

AIM: In order to explore the preventive mechanism of zinc, the changes of free radical signals and apoptosis in hepatic ischemia/reperfusion injury (HIR) rats were observed. METHODS: The MDA levels in serum were measured by fluorophotometry, free radical signals in liver were analyzed with electron spin resonance (ESR) method; and apoptosis was assayed by flow cytometry (FCM). RESULTS: MDA levels in serum and free radical contents in liver were both increased in rats with HIR. After zinc supplementation, they were decreased. After HIR, the percentage of subdiploid cells was 57.72% while it was reduced to 40.85% after zinc supplementation. CONCLUSION: Zinc may protect against HIR injury by inhibiting the production of free radicals in liver and hepatocellular apoptosis.

[Effect of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice].

This experiment mainly describes the effects of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice. The results were as follows: in acupuncture group, NK cell activity and T-lymphocyte positive rate of acid alpha-naphthyl acetate esterase (ANAE) and lymphocyte transformation rate were all increased. Compared with the control group, there was a significant difference (P < 0.01). The difference was insignificant, when compared with normal group (P > 0.05). Comparing the pathology grading of acupuncture group with control group, it showed marked difference in pathological section (P < 0.01). Adenoid structure and the degree of lymphocytic infiltration also have marked difference between acupuncture and control group (P < 0.05). Less tumour volume in acupuncture than control group were observed (P < 0.01). This indicated that acupuncture might increase the immunologic function of transplanted mammary cancer in mice and inhibit the growth of mammary cancer and enhance both differentiation level of mammary cancer cells and lymphocytic infiltration. Possibly acupuncture mightt reduce the malignancy of mammary cancer cells.

Changes in cellular immunity and nutritional status in mice after thermal injury.

A 13 per cent body surface area (BSA), full skin thickness burn was inflicted on LACA male mice and the changes in cellular immunity and nutritional status were observed. The results showed that thymus, spleen and circulating lymphocytes were significantly involved. A diminished mitogen responsiveness of spleen cells and altered peritoneal macrophage function were confirmed. Ear swelling tests indicated that the cellular immunity of burned mice was most severely depressed in week 2 postburn. The present study also showed that the dramatic change in nutritional status occurred earlier than that in cellular immunity and suggested the importance of early nutritional support after thermal injury.

Detection of epoxide hydrolase in rat hepatoma cell lines and primary rat liver cells by immunoblotting.

Rat epoxide hydrolase (EH) (EC 3.3.2.3) is elevated in cells of premalignant liver lesions, and variable EH activity has been reported for hepatocellular carcinomas. To facilitate detection of altered EH levels in liver cells, an immunoblotting method was devised. Rabbit antiserum specific for rat EH was prepared and used to detect EH extracted from suspensions of normal liver cells and from hepatoma cell lines. Compared with normal liver cells, 3 rat hepatoma cell lines, 7777, HTC and 17X, showed virtually undetectable EH levels by immunoblotting. The immunoblotting results rule out the possibility that very low EH enzymatic activity in the hepatoma cells results from production of normal amounts of non-functional enzyme protein.