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Objective:To investigate the changes of non-specific and HBV core antigen (HBcAg)-specific Th9 cells, and intereleukin-9 (IL-9) in HBV-infected patients, and to assess the influence of Th9 cells on CD8 + T cell function. Methods:Twelve patients with acute hepatitis B (AHB) and 58 with chronic hepatitis B (CHB), who were hospitalized in the First Affiliated Hospital of Xinxiang Medical University between January 2018 and January 2019, were enrolled in this study. Twenty healthy subjects negative for HBsAg were selected as controls. Peripheral blood mononuclear cells (PBMCs) and plasma samples were isolated. Non-specific Th9 cells (CD3 + CD4 + IL-9 + ) and HBcAg-specific Th9 cells were analyzed by flow cytometry. Plasma IL-9 level was measured by enzyme linked immunosorbent assay. CHB patients received tenofovir disoproxil fumarate (TDF) antiviral therapy. The changes of non-specific Th9 cells, HBcAg-specific Th9 cells and plasma IL-9 level were assessed 48 weeks after TDF therapy. CD4 + CCR4 -CCR6 -CXCR3 -(Th9) cells and CD8 + T cells were isolated from 12 HLA-A2 restricted CHB patients and co-cultured with HepG2.2.15 cells with the presence of anti-IL-9 neutralizing antibody. The percentage of dead HepG2.2.15 cells and the levels of IFN-γ and TNF-α were detected. Student′s t test, one-way analysis of variance or SNK- q test was used for statistical comparison between groups. Results:There were no significant differences in non-specific Th9 cells or plasma IL-9 level among AHB patients, CHB patients and healthy controls ( P>0.05). HBcAg-specific Th9 cells was down-regulated in CHB patients when compared with AHB patients [(2.49±0.61)% vs (3.19±0.62)%, P<0.001]. The percentage of HBcAg-specific Th9 cells was negatively correlated with HBV DNA ( r=-0.385, P=0.003), but not correlated with ALT ( P>0.05) in CHB patients. TDF therapy for 48 weeks remarkably elevated the HBcAg-specific Th9 cells [(2.94±0.48)%, P<0.001], however, did not affect non-specific Th9 cells or plasma IL-9 level ( P>0.05) in CHB patients. The cytotoxicity of HBcAg-specific Th9 cells was low in CHB patients. However, HBcAg-specific Th9 cells could induce enhanced cytotoxicity of CD8 + T cells to HepG2.2.15 cells, which manifested as increased percentage of dead HepG2.2.15 cells and higher levels of IFN-γ and TNF-α. Anti-IL-9 neutralizing antibody reduced the enhancement of CD8 + T cell cytotoxicity by HBcAg-specific Th9 cells ( P<0.001). Conclusions:Chronic HBV infection might suppress the level and function of HBcAg-specific Th9 cells, resulting in persistent infection.
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Objective:To investigate the expressions and significances of transforming growth factor β 1 (TGF-β 1) and dimethylarginine dimethylaminohydrolase 2 (DDAH2) in different stages of early-stage lung adenocarcinoma, and to provide basis for accurate pathological diagnosis of lung adenocarcinoma. Methods:Sixty-two surgical specimens from patients with early-stage lung adenocarcinoma in Shanxi Bethune Hospital from January 2012 to June 2019 were selected. Most of the specimens contained more than one pathological type. According to the pathological types, they were divided into three groups: 18 cases of atypical adenomatous hyperplasia (AAH), 60 cases of adenocarcinoma in situ (AIS), and 55 cases of invasive carcinoma (CA) component in minimally invasive adenocarcinoma (MIA) and lepidic predominant adenocarcinoma (LPA), and the wall attached growth patterns were selected as control group. Immunohistochemistry was used to detect the expressions of TGF-β 1 and DDAH2 proteins in different pathological types of lesions and control tissues, and the correlation among them was analyzed. Results:TGF-β 1 and DDAH2 proteins were expressed in normal lung tissue epithelial cells, and the positive rates were high [46.8% (29/62) and 98.4% (61/62)]. The positive rates of TGF-β 1 and DDAH2 proteins in epithelial cells of AAH, AIS and CA increased gradually, the positive rate of TGF-β 1 protein was 16.7% (3/18), 31.7% (19/60) and 70.9% (39/55), the positive rate of DDAH2 protein was 66.6% (12/18), 81.7% (49/60) and 90.1% (49/55), and the differences of positive rates among different pathological types were statistically significant (all P < 0.01). The positive rates of TGF-β 1 and DDAH2 protein in interstitial fibroblasts of normal lung tissue, AAH, AIS and CA increased gradually, the positive rate of TGF-β 1 protein was 11.3% (7/62), 61.1% (11/18), 72.3% (44/60), and 83.6% (46/55), the positive rate of DDAH2 protein was 0 (0/62), 22.2% (4/18), 65.0% (39/60), and 98.2% (54/55), and the differences of positive rates among different pathological types were statistically significant (all P < 0.01). In lung adenocarcinoma tissues, there was a positive correlation between the expression of TGF-β 1 in epithelial cells and the expression of DDAH2 in interstitial fibroblasts ( r = 0.221, P = 0.011). Conclusions:The expression trend of TGF-β 1 and DDAH2 proteins in different pathological types of early-stage lung adenocarcinoma lesions may relate to the degree of lesions. The combined detection of TGF-β 1 and DDAH2 proteins is expected to be a biomarker for the auxiliary diagnosis of early-stage lung adenocarcinoma with different pathological types.
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Objective@#To analyze the clinical characteristics of oral candidiasis (OC) in in-patients with rheumatism, in order to provide theoretical basis for the prevention and treatment of OC in rheumatism patients.@*Methods@#One thousand eight hundred and eight in-patients were recruited in the Department of Rheumatology, the Second Hospital of Shanxi Medical University from January 2017 to December 2017. The patients included 607 males and 1 201 females. Their average age was (49.5±15.5) years old with a ranging from 14 to 81 years. According to occurrence of OC or not, the patients were divided into OC group and non-OC group. The differences of general data, primary diseases, laboratory examinations, usage of glucocorticoid and immunosuppressant therapy were compared between the two groups, and the risk factors of OC occurrence were analyzed.@*Results@#Two hundred and sixty-nine patients had OC and 1 539 patients had no OC. Age [(54.9±14.7) years], duration of illness [(9.4±4.4) years] and hospital stay [(15.3±5.7) d] in OC group were significantly longer than those in non-OC group. OC incidence in patients with connective tissue disease (CTD) [17.40% (193/1 109)] was higher than that in non-CTD patients [10.87% (75/699)] (P<0.001). OC most likely occurred in patients with such CTD as Sjögren syndrome (SS) and Behcet syndrome. OC incidence in non-CTD patients with osteoarthritis (OA) was highest. The salivary flow rate in OC group [(0.65±0.45) ml/min] was significantly lower than that in non-OC group [(0.78± 0.39) ml/min] (t=2.394, P=0.017). There was no statistical differences in other laboratory examinations between the two groups, including white blood cells (WBC), lymphocyte, platelet count, liver function, renal function, erythrocyte sedimentation rate, C-reactive protein, procalcitonin, immunoglobulin G, immunoglobulin M, immunoglobulin A, C3, C4 and so on. OC incidence in patients using prednisone≥15 mg/d [17.16% (133/775)] was higher than that in patients using prednisone<15 mg/d [12.53% (94/750)] and patients not using prednisone [14.84% (42/283)] (P<0.05). The incidence of OC in patients with immunosuppressant therapy [16.11% (226/1 403)] was statistically higher than that in non-immunosuppressant patients [10.62% (43/405)] (P<0.01). Logistic regression analysis showed that the risk factors of OC occurrence included primary diseases (P<0.001), age (P<0.001), duration of illness (P=0.001) and duration of hospitalization (P=0.002).@*Conclusions@#OC occurred commonly in rheumatism in-patients, especially in elder patients, patients with long duration of illness and hospital stay. OC incidence in CTD patients is significantly higher than that in non-CTD patients. Glucocorticoid and immunosuppressant therapy might significantly reduce the anti-fungal immunity of the patients.
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OBJECTIVE： To establish HPLC fingerprints of Nauclea officinalis extract syrup， and to determine the contents of 9 components. METHODS： HPLC method was adopted. The determination was performed on Diamonsil C18（2）column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using strictosamide as reference， HPLC chromatograms of 20 batches of N. officinalis extract syrup were drawn. The similarity of HPLC chromatograms were evaluated by using TCM Fingerprint Similarity Evaluation System （2004A edition） to confirm common peaks. The contents of 9 components were determined by standard curves. RESULTS： There were 26 common peaks in 20 batches of HPLC chromatograms， and the similarity was higher than 0.98. Compared with mixed control， 9 chemical components were identified， such as 3，4-dihydroxybenzoic acid， neochlorogenic acid， loganic acid， chlorogenic acid， cryptochlorogenic acid， swertioside， pumiloside， strictosamide and vincosamide. The linear range of 9 components were 17.24-275.84， 7.56-120.96， 15.40-246.40， 7.84-125.44， 8.64-138.24， 7.96-127.36， 8.40-134.40， 48.56-776.96， 4.16-66.56 μg/mL（all r≥0. 999）， respectively. The limits of detection were 0.043 1， 0.126 0， 0.038 5， 0.130 7， 0.144 0， 0.066 3， 0.070 0， 0.012 1， 0.052 0 μg/mL， respectively. The limits of quantitation were 0.215 5， 0.189 0， 0.077 0， 0.196 0， 0.288 0， 0.132 7， 0.105 0， 0.097 6， 0.138 7 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 99.6％、106.3％、100.1％、102.0％、98.4％、100.0％、99.3％、100.6％ and 101.2％， and RSDs were 1.20％、0.24％、0.59％、1.00％、0.73％、1.30％、1.10％、1.80％、1.90％（n＝6）. CONCLUSIONS： Established HPLC fingerprints and quantitative determination method of N. officinalis extract syrup are accurate， specific and sensitive. It can provides reference for quality control of N. officinalis extract syrup.
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Objective To observe the local field potential activity of dorsomedial and dorsolater striatal (STR) neurons in rats with exercise-induced fatigue,so as to explore the regional characteristics of those neurons.Methods Thirty-six male Wistar rats were randomly divided into a control group(CG),a single fatigue group(SFG)and a repeated fatigue group(RFG),each of 12.The exercise-induced fatigue model was established according to a load-increasing treadmill running protocol.The CG did not do any treadmill running,SFG finished one-time exhaustive exercise while the RFG repeated exhaustive exercise 6 times.The extracellular glass microelectrode technique was used to record the spontaneous firing of dorsomedial(n=6)and dorsolateral(n=6)STR neurons in vivo to observe the discharging frequency,characteristics and types.Results (1)Spontaneous firing frequencies of striatal neurons in SFG were significantly higher than those of CG(P＜0.01),while those of RFG were significantly lower than SFG.(2) After repeated fatigues,there were significant decreases in the irregular firing pattern and significant increase in the explosive firing of striatal neurons compared with CG(P＜0.05).(3)After the single fatigue and repeated fatigue,the discharging frequency of dorsomedial medium spiny-like neurons increased significantly(P＜0.05),and that after repeated fatigue significantly higher than that of the dorsolateral(P＜0.01).The discharge frequency of the dorsomedial and dorsolateral fast-spiking neurons decreased significantly after repeated fatigues.The discharge frequency of dorsolateral large aspiny-like neurons of RFG was significantly higher than SFG,while that of the latter was significantly lower than CG(P＜0.05).Conclusions (1)Striatal neurons mediate exercise-induced fatigue and present regional characteristics,which might be due to dorsomedial and dorsolateral striatal neurons receiving different types of projection neurons.(2) Striatal fast-spiking neurons may play an important role in mediating the exercise-induced fatigue.
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Objective To explore the value of hysteroscopy in diagnostic curettage, the incidence of complications and preventive measures in senile women. Methods One hundred and thirteen senile patients who underwent hysteroscopy from January 2002 to December 2007 were recruited in the study, and one hundred and fifty-seven non senile patients were selected as control group. All the patients were operated with diagnostic curettage and hysteroseopy. The success rate of hysteroscopy operation, the incidence of complications and control effect were recorded and compared between two groups. Results The success rate of hysteroscopy operation was 88. 5% (100/113) in the senile group and 100% (157/157) in the non senile group, and there was significant differenee(P <0. 01). The incidence of complications was higher in the senile women group than that in the non senile women group (P< 0. 01). In senile group, the accuracy of diagnostic curettage and hysteroscopy was higher than diagnostic curettage alone, and the success rate was 95.5% (64/67)in estrogen group and 69.6%(32/46) in blank control group (P<0. 01) ,which indicated that estrogen was positively related with the success rate of operation. Compared with misoprostol, estradiol-pretreatment operation was safer and had higher success rate. Conclusions It is safe and feasible to perform hysteroscopy and diagnostic curettage for older women, though it has higher incidence of complications and lower success rate compared with non senile women. Hysteroscopy and diagnostic curettage are more accurate than diagnostic curettage alone, and it is a good choice to apply estradiol before operation.
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BACKGROUND: Studies have shown that alanine (A) to threonine (T) substitution at codon 54 of intestinal fatty acid-binding protein (FABP2) in different populations is associated with dyslipidemia and other characteristics of metabolic syndrome.OBJECTIVE: To investigate the frequency of encoding 54Ala/Thr (A/T) single nucleotide polymorphism in the FABP2 in middle-aged and old people, and explore the association between 54T FABP2 and plasma lipids.DESIGN: A case-controlled analysis. SETTING: Department of Biochemistry, Hebei North University and Department of Clinical Laboratory, the 251 Hospital of Chinese PLA.PARTICIPANTS: 469 physical examinees were selected from the Medical Examination Center, the 251 Hospital of Chinese PLA between October 2003 and April 2005. The subjects included 217 males with mean age of (56±10) years, and 252 females with mean age of (55±13) years. Only people with normal liver and kidney function, and with no blood relation were recruited. The informed consent to this study was obtained from all subjects. The experiment was admitted by Hospital Ethics Committee. METHODS: ①After fasting for 12 hours, automatic analyzer (Olympus AU 6400) was adopted to measure plasma total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apoprotein A1(Apo A1) and Apo B levels. ②1 mL venous blood was extracted and immediately mixed with anti-coagulants containing citric acid, natrium citricum and glucose. White blood cells were separated and genomic DNA was isolated using standard methods with proteinase K digestion and phenol/chloroform purification. The genotype distribution frequency in each group was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). MAIN OUTCOME MEASURES: ①Plasma TC, TG, HDL-C, LDL-C, Apo A1and Apo B levels; ②Distributions of FABP2 genotypes at codon 54. RESULTS: ①The genotype frequencies of A/A, A/T, T/T were 0.48, 0.42, and 0.10 in males, and 0.44, 0.46, and 0.10 in females, respectively. The allelic frequency of point mutant 54Thr in FABP2 gene was 0.31 in males and 0.33 in females, respectively. There was no difference between males and females (χ2=0.47, P ＞ 0.05). ②The LDL-C and Apo B concentrations in fasting plasma of males with 54T allele were significantly higher than those with 54A allele (P ＜ 0.05). The TC and LDL-C concentrations in fasting plasma of females with 54T allele were significantly higher than those with 54A allele (P ＜ 0.05). CONCLUSION: In the middle-aged and old populations, the frequency of encoding 54Ala/Thr polymorphism in FABP2 gene is not correlated with gender, but with high lipoprotein profile.