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Objective:To find the age-related indicators of human immune function in the blood, to explore the possibility to standardize the evaluation criterion of immune function in Chinese population.Methods:The peripheral blood samples from 478 healthy individuals with complete medical data collected at the Beijing Hospital from January 2019 to December 2019, were chosen and analyzed.The volunteers were divided into young(<40 years old)and elderly(≥70 years old)groups.The differences in RNA, cytokines and immune cell function(the proliferation of T cell as well as the ability of T cells and natural killer cells to lyse target cells)were compared between the young group versus the elderly group.Results:Real-time quantitative fluorescence RT-PCR detection method was used for detecting the following tumor immune-related genes in the peripheral blood of healthy people in young versus old groups: programmed death 1( PD1)gene, complement C3 gene( C3), complement C4 gene( C4), high sensitivity C-reactive protein gene( HsCRP), tumor necrosis factor alpha gene( TNFA), interferon gamma gene( INF- γ), interleukin-6 gene( IL-6), interleukin-8 gene( IL-8), cytotoxic T lymphocyte-associated antigen-4 gene( CTLA-4), nuclear transcription factor-κB gene( NF- κB), long intergenic noncoding RNA-erythroid pro-survival gene( lincRNA- EPS), long non-coding RNA-myeloid RNA regulator of Bim-induced death gene( lncRNA- Morrbid), lincRNA-cyclooxygenase 2 gene( lincRNA- Cox2), Lnc-Dendritic cells( Lnc- DC), and circRNA-7 gene( ciRS-7). The mRNA level of C4 was significantly lower in elderly people than in young controls[young group(1.01±0.18) vs.old group(0.64±0.13), P=0.047], while others showing a non-statistically significant difference( P>0.05). The protein levels of programmed death 1(PD1), complement C3(C3), complement C4(C4), high sensitivity C-reactive protein(HsCRP), tumor necrosis factor alpha(TNF-α), interferon gamma(INF-γ), interleukin-6(IL-6), interleukin-8(IL-8), cytotoxic T lymphocyte-associated antigen-4(CTLA-4)and nuclear transcription factor-κB(NF-κB)in plasma were detected by enzyme-linked immunosorbent assay(ELISA). The following protein levels were significantly higher in the elderly group than in the young group: IL-6 protein[young group(249.30±100.02)ng/L vs.old group(283.00±94.98)ng/L, P=0.021], HsCRP protein[young group(2543.00±1111.89)ng/L vs.old group(3056.00±1056.61)ng/L, P=0.002], INF-γ protein[young group(362.40±383.67)ng/L vs.old group(500.40±502.27)ng/L, P=0.047]and TNF-α protein[young group(20.74±29.47)ng/L vs.old group(33.09±48.91)ng/L, P=0.042]. While the level of C4 was significantly lower in the elderly group than in the young group[young group(449.50±51.17)ng/L vs.old group(407.10±59.78)ng/L, P<0.0001]. T-cell proliferation, quantified by flow cytometry-based fluorescent dye dilution, showed that the CD3 + T cells of elder people had proliferated through 4 generations, while the CD3 + T cells of young people had proliferated through 5 generations.As shown in the NK killing assays, the secretion of LDH in the target A549 tumor cells decreased significantly after treated with NK cells belonging to elder individuals.The result demonstrated that individuals over 70 years old have significantly lower levels of the killing activity of NK cells( P<0.05). However, there was not statistically significant in T lymphocyte killing activity between different ages( P>0.05). Conclusions:IL-6, HsCRP, INF-γ and TNF-α levels are increased with aging, while the level of C4 significantly is decreased.The proliferation ability of T cells and the ability of NK cells to kill tumor cells are weakened.
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Objective To investigate activation of the phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(Akt)pathway in the endometrium of women with polycystic ovary syndrome(PCOS)and its role in endometrium hyperplasia and carcinogenesis,and the factors affecting the activation of the PI3K/Akt pathway.Methods From Jan 2007 to Jun 2008,52 patients with PCOS who underwent dilatation and curettage were selected as experimental group matched with 32 non-PCOS patients as control group.Serous hormonal parameters,fasting blood glucose and insulin,body mass index(BMI),and endometrium pathology were measured and evaluated in all patients.The PCOS patients were divided into insulin resistance and non-insulin resistance group according to homeostasis model assessment-insulin resistance index (HOMA-IR).Meanwhile,the PCOS patients were grouped as normal,endometrial hyperplasia and carcinoma depending on outcome of pathology.The expression of Akt and phosphorylated Akt(p-Akt)were determined by western blot.Results(1)The expression of p-Akt was significantly higher in PCOS group [(46±18)％]than that in control[(33 ±9)％,P ＜0.01)].(2)The expression of p-Akt was significantly higher in group of endometrial hyperplasia and carcinoma[(56 ± 19)％]when compared with those in normal endometria group[(31 ± 12)％,P ＜ 0.05]; the expression of p-Akt was significantly higher in group of insulin resistance[(50 ± 19)％]compared with that in non-insulin resistance group [(34 ± 10)％,P ＜0.01].(3)There was a positive correlation between the expression level of p-Akt in endometrium with PCOS and HOMA-IR and BMI respectively(r =0.400,0.326,both P ＜ 0.05).Conclusions The PI3K/Akt pathway was over activated in endometrium with PCOS which may be associated with the formation of endometrial hyperplasia and carcinoma in PCOS patients.Insulin resistance and obesity may be high risk factors for over-activation of the PI3K/Akt pathway in endometrium with PCOS.
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Objective To investigate the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling pathway in the endometrium of women with polycystic ovary syndrome (PCOS) and its effect and significance in the cause of hyperplasia and carcinoma;and investigate the factors which affect the activation of the MAPK/ERK signaling pathway. Methods Collected 52 patients diagnosed as PCOS who were taken dilation and curettage of uterus as study, while 32 non-PCOS patients matched as control group. Serum hormonal parameters, fasting blood glucose and insulin were measured in all patients. The PCOS patients were sub-group as insulin resistance group and non-insulin resistance group; all the patients were carried out pathology inspection of endometria, and the PCOS patients were sub-group as endometrial hyperplasia and carcinoma group and normal endometrium group based on the outcome of pathology inspection. Western blot were performed to detect the expressions of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2), the activation of ERK1/2. Results (1)The expression of pERK1/2 [(61 ±13)%] in the endometrium in PCOS group was higher than that in the control [(44 ±10)%, P ＜0.01]. (2)The expression of p-ERK1/2 was significantly increased in group of endometrial hyperplasia and carcinoma [ ( 70 ± 11 )% ] compared to that in group of normal endometrium [ (55 ± 10)% ,P ＜ 0.01 ], while there were significant difference between group of insulin resistance [ (63 ± 13 )% ] and group of non-insulin resistance [ (55 ±7)%, P ＜0.01 ]. (3) Fasting insulin level, insulin area under the curve and body mass index were related to the expression of p-ERK1/2 in endometrium with PCOS, the correlation coefficient were 0.447, 0.456 and 0.381, respectively ( all P ＜ 0.01 ). Conclusions The MAPK/ERK signaling pathway in endometrium with PCOS was overactivation, which was related to the endometrial hyperplasia and carcinoma; while the activation of MAPK/ERK signaling pathway were effected by insulin resistance and hyperinsulinemia.
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Objective To discuss the significance of near infrared spectroscopy (NIRS) in evaluation of intrapartum hypoxic-ischemic cerebral injury, and to provide a method to evaluate neonatal brain damage objectively and quantitatively. Methods A total of 63 neonates with fetal distress were divided into hypoxic-ischemic encephalopathy(HIE) group and non-HIE group. Thirtyfive newborns with no fetal distress were chosen as controls. Using NIRS, the brain regional oxygen saturation(rSO2) in these neonates were measured. Evaluation of brain rSO2 in the diagnosis of HIE was analyzed with receiver operating characteristic (ROC) curve. Results At the time of fetal head visible on vulval gapping and 5 min after birth, the HIE group showed decreased brain rSO2[(36. 6±5.0)% and (52. 0±4. 2)%], comparing with control group[(45. 9±4. 6)% and (59. 6±4. 4)%]and non-HIE group[(44.1±3.1) % and (57. 6±3. 5) %](P<0. 01) . The brain rSO2 was positively correlated with the pH and oxygen saturation of umbilical artery blood in all groups (P<0. 01). When the cut-off value of brain rSO2 was <39. 5% at fetal head visible on vulval gapping, the sensitivity and specificity of assessing HIE were 67% and 93%, respectively, while 70% and 86% when the cut-off value was <53. 5% at 5 min after birth. Conclusions The brain rSO2 obtained by NIRS could be used to evaluate brain oxygenation, and may be useful in predicting HIE in neonates with fetal distress.
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Objective To investigate the function of metalloproteinase 2 (MMP 2) and tissue inhibitor of metalloproteinase 2 (TIMP 2) in neonatal acute lung injury(ALI) caused by LPS in rats. Methods Eighty eight newborn rats of 7 days were randomly divided into eight groups: control, 0.5 h, 1 h, 2 h, 4 h, 8 h, 16 h, 24 h after LPS was injected. The changes of lung pathology in newborn rats were observed and the changes of MMP 2 and TIMP 2 expression were measured by immunohistochemistry and RT PCR. Results Pulmonary hemorrhage was seen in newborn rats caused by LPS. The expression of MMP 2 mRNA and protein were 0.523?0.030 and 126.20?17.98; The expression of MMP 2 mRNA increased and the highest level was at 4 h(0.826?0.567, t=3.77, P
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Objective To explore the role of caspase-3 activation and DNA fragmentation in later period of neonatal rat hypoxic-ischemic brain damage(HIBD). Methods DNA fragmentation,caspase-3 mRNA and caspase-3 protein were measured in 2 wks、3 wks and 4wks after setting HIBD animal model in newborn wistar rats by FCM, RT-PCR and Immunohistochemistry. Results (1) Apoptosis lasted 4ks after HIBD. This suggested a long lasting role of apoptosis in neonatal HIBD by TNNEL and EM.(2)Caspase-3 mRNA expression was estimated by semi-quantitative RT-PCR. It was higher in HIBD group(0.771?0.074) than in control group(0.620?0.038, P0.05. Average Avalue of Caspase-3 protein in HIBD group(0.374 ?0.038) at 3 wks was significantly higher than that in control group(0.356?0.020,P
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<p><b>OBJECTIVE</b>To investigate the effect of second messengers protein kinase C (PKC) and inositol (1,4,5)-trisphosphate (IP(3)) during hypoxic-ischemic brain injury in neonatal rats.</p><p><b>METHODS</b>The protein concentration was determined by Lowry's method. PKC activity was measured by the incorporation of [gamma-32P] into a specific substrate peptide in the cytosol and particulate fraction, respectively. IP(3) was determined by the radioreceptor binding assay.</p><p><b>RESULTS</b>Compared with the control, PKC activities in the particulate fractions in both the cerebral cortex and hippocampus decreased, while increased in the cytosol in cerebral cortex and remained within the normal range in the cytosol in the hippocampus at 0, 4, 12, 24, 48, 72 h, 7, and 14 d after hypoxic-ischemia for 20 min. All these changes restored to normal levels at 21 d post hypoxic-ischemia. Similarly, a decrease in IP(3) in the cerebral cortex and hippocampus and an increase in IP(3) in the thalamus after hypoxic-ischemia were noted, respectively. Changes in cytosolic PKC activity were not related to those of IP(3), as evaluated statistically.</p><p><b>CONCLUSION</b>Hypoxia-ischemia induces disturbance of the second messengers IP(3) and PKC, which may play important roles in the pathogenesis of hypoxic-ischemic brain injury.</p>
Subject(s)
Animals , Female , Male , Rats , Animals, Newborn , Brain Ischemia , Metabolism , Hypoxia, Brain , Metabolism , Inositol 1,4,5-Trisphosphate , Physiology , Protein Kinase C , Physiology , Rats, WistarABSTRACT
Objective To investigate the mechanism of intracellular calcium and other ions disturbance by measuring the activity of Ca 2+ adenosine triphosphatase (Ca 2+ ATPase) and Na + K + adenosine triphosphatase (Na + K + ATPase) Methods Model of fetal rats ischemia and reperfusion was established The duration of ischemia was 15,30,45 and 60mins respectively;after ischemia for 15 mins, reperfusion for 1,4,8,15 and 24 hours There were 7 11 fetal rats sacrificed at different time points respectively, 12 rats in sham for control The mitochondria and endoplasmic reticulium (microsomia) were estracted and the activity of the enzyme was measured Results In the ischemia group: with the development of ischemia, the activity of Ca 2+ ATPase in mitochondria decreased gradually ( P
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Objective To study the expression of ICAM 1 at gene transcription level in neonatal rats after hypoxic ischemic brain damage(HIBD). Method Intercellular adhesion molecule 1(ICAM 1) mRNA was detected in cortex of neonatal rats at different times after HIBD and control by reverse transcription polymerase chain reaction(RT-PCR) technique. Result In the ischemic cortex, levels of ICAM 1 mRNA increased markedly at 6 h(2.5 fold, t=2.33, P
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AIM To analyze the mechanism of the adriamycin induced cardiomyopathy. METHODS The patch clamp technique in the whole cell recording was used to study the effect of adriamycin on L type calcium channel current( I Ca L ) in the isolated cardiomyocyte of the guinea pig. RESULTS The current voltage( I U ) curveshowed the bell shape in the control and in 0 1 mmol?L -1 adriamycin. Their peak potentials were about +10 mV. The amlitude of peak calcium current increased from (-0 93?0 05) nA to (-1 31?0 08) nA( P