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Objective: To observe the effects of electroacupuncture (EA) pretreatment on M1 polarization of alveolar macrophages (AMs) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS), and to explore the potential protective mechanism of EA.Methods: Forty Sprague-Dawley rats were randomly divided into a normal group, a model group, and three groups of EA pretreatment [including a Chize (LU5) group, a Zusanli (ST36) group and a Chize (LU5) plus Zusanli (ST36) group], with eight rats in each group. The model rats of ALI were established by instilling LPS [2 mg/(kg·bw)] into the trachea of rats for 3 h. The rats in each EA pretreatment group were pretreated with EA for 30 min per day at the corresponding bilateral acupoints 6 d before instilling LPS. Three hours after modeling, the pulmonary function of the rats was tested, and the lung tissue was taken to calculate the ratio of lung wet weight to dry weight (W/D). The pathological lung changes and the injury score were observed by hematoxylin-eosin staining. The contents of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and myeloperoxidase (MPO) in rat's bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay. The mRNA and protein expression levels of M1 macrophage markers clusters of differentiation 86 (CD86), inducible nitric oxide synthase (iNOS), and its signaling pathway factor Toll-like receptor (TLR) 4, and nuclear factor-κB (NF-κB) p65 in the alveoli were detected by fluorescence quantitative polymerase chain reaction and Western blot, respectively. Results: After being induced by LPS, the pulmonary function of the model rats showed that the forced expiratory volume in 0.1 s (FEV0.1), forced expiratory volume in 0.3 s (FEV0.3), and their respective ratios of FEV to forced vital capacity (FVC) (including FEV0.1/FVC and FEV0.3/FVC) were significantly decreased (P<0.01), while the W/D of lung tissue was increased (P<0.01). The score of lung injury was significantly higher (P<0.01). The contents of TNF-α, IL-1β, and MPO in the BALF and the mRNA and protein expression levels of CD86, iNOS, TLR4, and NF-κB p65 in the lung tissue were significantly increased (P<0.01). After EA pretreatment, the FEV0.1, FEV0.3, FEV0.1/FVC, and FEV0.3/FVC were significantly increased, the lung injury score decreased significantly, and the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs and proteins in the alveoli decreased significantly (P<0.05 or P<0.01). Compared with the other two single acupoint groups, the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs in the alveoli in the Chize (LU5) plus Zusanli (ST36) group were significantly lower (P<0.01). Conclusion: EA pretreatment at Chize (LU5) and Zusanli (ST36) can inhibit inflammation and reduce pulmonary injury in ALI rats induced by LPS. The effect of the combination of Chize (LU5) and Zusanli (ST36) is better than that of using these two acupoints separately, and its mechanism may be related to the inhibition of AMs' M1 polarization by down-regulation TLR4/NF-κB signaling pathway.
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Endogenous ribonucleotides(RNs)and deoxyribonucleotides(dRNs)are important metabolites related to the pathogenesis of many diseases.In light of their physiological and pathological significances,a novel and sensitive pre-column derivatization method with N-(t-butyldimethylsilyl)-N-methyltri-fluoroacetamide(MTBSTFA)was developed to determine RNs and dRNs in human cells using high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS).A one-step extraction of cells with 85％methanol followed by a simple derivatization reaction within 5 min at room temper-ature contributed to shortened analysis time.The derivatives of 22 nucleoside mono-,di-and tri-phosphates were retained on the typical Cig column and eluted by ammonium acetate and acetonitrile in 9 min.Under these optimal conditions,good linearity was achieved in the tested calibration ranges.The lower limit of quantitation(LLOQ)was determined to be 0.1-0.4 μM for the tested RNs and 0.001-0.1 μM for dRNs.In addition,the precision(CV)was＜15％and the RSD of stability was lower than 10.4％.Furthermore,this method was applied to quantify the endogenous nucleotides in human colorectal carcinoma cell lines HCT116 exposed to 10-hydroxycamptothecin.In conclusion,our method has proven to be simple,rapid,sensitive,and reliable.It may be used for specific expanded studies on intracellular pharmacology in vitro.
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Objective:To use quantitative computed tomography (QCT) technology to measure the bone mineral density of the spine of the Chinese healthy population, and to explore its correlation with hemoglobin and serum albumin.Methods:The data in this study came from the China Health Quantitative CT Big Data Project (China Biobank). The spine bone density was measured by using QCT Pro Image Analysis System and all cooperating centers used the European spine phantom (NO.145) for quality control. Total of 50 053 healthy persons who met the criteria for entry were selected as the research subjects. The subjects were divided into 7 groups according to age. The general data, spine bone density, serum albumin, hemoglobin of the subjects were collected. The single-factor analysis of variance, Pearson correlation analysis and multi-classification logistic regression model were applied to analyze the correlation between bone density and hemoglobin and serum albumin.Results:The bone mineral density of healthy people decreased with age ( P<0.05), and there were significant differences in hemoglobin, serum albumin and body mass index (BMI) among different age groups (all P<0.05). Linear correlation analysis showed that there were positive correlation between bone mineral density and hemoglobin in healthy males in different age groups ( r=0.086, 0.101, 0.076, 0.090, 0.072, 0.123, 0.100, all P<0.01). There were negative correlation between bone mineral density and hemoglobin in certain age groups in women (40-49 years group: r=-0.027; 70-79 yearsgroup: r=-0.077; both P<0.05). And corelation were found between bone mineral density and serum levels of albumin in certain age groups of healthy subjects (among men, 30-39 years group: r=-0.048; 40-49 years group, r=-0.027; 70-79 years group, r=-0.051; among women, 30-39 years group: r=-0.044; 40-49 years group, r=-0.042; 50-59 years group, r=-0.086; 70-79 years group, r=-0.070; all P<0.05). After adjusting for age and BMI, the multi-category logistic regression analysis showed that the hemoglobin level was protective factor of normal bone density ( OR=1.022, 95% CI:1.017-1.027) and decreased bone density ( OR=1.012, 95% CI:1.007-1.016) in healthy males, and the serum albumin was risk factor for normal bone density ( OR=0.926, 95% CI:0.905-0.948) and decreased bone density ( OR=1.006, 95% CI:0.951-1.011) in healthy women. Conclusion:There is a correlation between bone mineral density and hemoglobin and serum albumin in Chinese healthy population. Hemoglobin is a protective factor for bone mineral density in men, and serum albumin is a risk factor for bone mineral densityin women.
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Objective:To investigate the clinical and epidemiological features of acute respiratory adenovirus infection in children.Methods:Clinical data of 488 children with acute respiratory tract human adenovirus (HAdV) infection admitted in Children’s Hospital of Zhejiang University School of Medicine from September 2018 to August 2019 were retrospectively analyzed. Nasopharyngeal swabs or nasopharyngeal aspirates (NPAs) were collected and tested by direct immunofluorescence assay. Kruskal Wallis H test was used for quantitative data without normal distribution, and P<0.05 was considered to be statistically significant. The qualitative data were compared by chi-square test or Fisher’s exact test. Bonfereoni chi-square segmentation was performed for comparison between groups, and P<0.007 was considered statistically significant after correction. Results:A total of 488 HAdV positive cases were detected from 7 072 patients with acute respiratory tract infection (6.9%), including 305 males (62.5%) and 183 females (37.5%). The median age of HAdV positive children was 43 months (39 days to 12 years). The detection rate in 6 m-<2 y age group(8.7%, 123/1 408)was significantly higher than those in <6 m group (3.0%, 6/197)and ≥5 y group(4.6%, 89/1 948)( χ2=7.57, 23.98, P all <0.007). The detection rate in 2-<5 y group(7.7%, 270/3 519)was significantly higher than those in <6 m group and ≥5 y group ( χ2=5.809, 19.688, P all <0.007). The peak rate was detected in the winter [12.9%(238/1 840)] which was significantly higher than those in spring (4.7%), summer (3.9%), and autumn(5.5%)( χ2=103.477, 58.986 and 49.926, P<0.007). The average length of hospital stay was (6±4)d(1-41 d). 486 cases (99.5%) were discharged from hospital after treatment, and 2 cases died. There were 111 cases (22.7%) of acute upper respiratory tract infection, 34 cases (7.0%) of bronchitis and 343 cases (70.3%) of pneumonia; and severe pneumonia was diagnosed in 86 cases (25.1%, 86/343). The common clinical manifestations were fever 93.4% (456/488), cough 94.7% (462/488), wheezing 26.2% (128/488) and shortness of breath 14.8% (72/488). 138 cases (28.3%) had extrapulmonary symptoms, 78 cases (16.0%) had underlying diseases, among which congenital heart disease was most common (16, 3.3%). The average duration of fever was(8.8±2.4)d(5-17 d)in 456 fever cases, the duration between 7-10 d in 277 cases and >10 d in 96 cases; and 439 cases had hyperpyrexia(≥39 ℃). The single infection occurred in 275 (56.4%) cases and mixed infection in 213(43.6%) cases. The proportions of fever, hyperpyrexia, fever duration >10 d, severe pneumonia, wheezing and length of hospital stay in mixed infection group were significantly higher than those in the single infection group ( χ2/ Z=11.960, 6.494, 37.209, 72.841 and -8.805, P all <0.05). The length of hospital stay, proportion of fever time>10 d, wheezing, shortness of breath, hypersomnia/poor spirits, serous effusion, extrapulmonary symptoms, mixed infection, and underlying diseases in severe pneumonia group were significantly higher than those in the mild pneumonia group ( χ2/ Z=-9.182, 23.825, 49.094, 143.627, 219.659, 81.327, 8.080, 21.546 and 10.556, P all <0.05). The proportion of severe pneumonia in 6 m to <2 y group was higher than that in 2-<5 y group and ≥5 y group( χ2=20.709, 8.603, P all <0.007). Conclusions:HAdV is an important pathogen of acute respiratory infection in children. HAdV infection occurs mainly in children aged from 6 month to 2 years and has a high detection rate in winter. Children aged 6 months to 2 years with wheezing, shortness of breath, underlying diseases, extrapulmonary symptoms and mixed infections are more likely to develop severe pneumonia.
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Objective@#To understand clinical characteristics of children with pneumococcal meningitis (PM) in China and to analyze the drug sensitivity of Streptococcus pneumoniae isolates and associated impacts on death and sequelae.@*Methods@#The clinical data, follow-up results and antimicrobial sensitivity of isolated strains of 155 children (including 98 males and 57 females, age ranged from 2 months to 15 years) with PM in 10 tertiary-grade A class hospitals of Infectious Diseases Surveillance of Pediatrics (ISPED) from 2013 to 2017 were collected and analyzed retrospectively. Patients were divided into different groups according to the following standards: ≤1 year old group,>1-3 years old group and >3 years old group according to age; death group and non-death group according to the death within 30 days after PM diagnosis; complication group and non-complication group according to the abnormal cranial imaging diagnosis; sequelae group and no-sequelae group according to the follow-up results. Bonfereoni chi-square segmentation and Kruskal-Wallis H test were used for statistical analysis.@*Results@#There were 64 cases (41.3%) in the ≤1 year old group, 39 cases in the >1-3 years old group (25.2%), and 52 cases (33.5%) in the >3 years old group. The most common clinical manifestation was fever (151 cases, 97.4%). The mortality was 16.8% (26/155) during hospitalization. The neurological complication rate was 49.7% (77/155) during hospitalization, including the most common complication, subdural effusion and (or) empyema in 50 cases (32.3%) and hearing impairment in 6 cases. During follow-up after discharge, no death was found and focal neurological deficits were found in 47 cases (30.3%), including the frequent neurological sequelae: cognitive and mental retardation of different degree in 22 cases and hearing impairment in 14 cases (9.0%). The rate of cure and improvement on discharge was 74.8% (116/155) and the lost to follow-up rate was 8.4% (13/155). The proportions of died cases, neurological complications during hospitalization and proportions of peripheral white blood cell count <12 × 109/L before admission in ≤1 year old group were significantly higher than those in >3 years old group (25.0% (16/64) vs. 5.8% (3/52), 75.0% (48/64) vs. 25.0% (13/52), 48.4% (31/64) vs. 15.4% (8/52), χ2=7.747, 28.767, 14.044; P=0.005, 0.000, 0.000). The proportions of headache, vomiting, neck resistance and high risk factors of purulent meningitis in >3 years old group were significantly higher than those in ≤ 1 year old group (67.3%(35/52) vs. 1.6%(1/64), 80.8% (42/52) vs. 48.4% (31/64), 69.2% (36/52) vs. 37.5% (24/64), 55.8% (29/52) vs. 14.1%(9/64), χ2=57.940, 12.856, 11.568, 22.656; P=0.000, 0.000, 0.001, 0.000). Streptococcus pneumoniae isolates were completely sensitive to vancomycin (100.0%, 152/152), linezolid (100.0%, 126/126), moxifloxacin (100.0%, 93/93) and ofloxacin (100.0%,41/41); highly sensitive to levofloxacin (99.3%, 142/143) and ertapenem (84.6%, 66/78); moderately sensitive to ceftriaxone (48.4%, 45/93), cefotaxime (40.0%, 44/110) and meropenem (38.0%, 38/100); less sensitive to penicillin (19.6%, 27/138) and erythromycin (4.2%, 5/120). The proportions of non-sensitive strains of penicillin (21/21) and meropenem (17/18) in the death group were significantly higher than those (90/117, 45/82) in the survived group(χ2=4.648 and 9.808, P=0.031 and 0.002).@*Conclusions@#The children′s PM is mainly found in infants under 3 years old in China. Death and neurological complications are more common in PM children under 1 year old. The clinical manifestations and peripheral blood inflammatory markers of PM patients under 1 year old are not typical. Fever is the most common clinical manifestation and subdural effusion and (or) empyema is the most common complication. Long-term hearing impairment is common in PM and the follow-up time must be prolonged. The dead PM cases had high in sensitive rates to penicillin and meropenem.
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Objective To explore the clinical value of real-time contrast-enhanced ultrasonography and ultrasonography in the diagnosis and staging of cervical cancer.Methods 80 cervical cancer patients were selected as the research subjects,they were randomly divided into two groups.40 patients in the observation group received real time contrast-enhanced ultrasound,40 cases of the control group received conventional ultrasonography.The detection rate,diagnosis of cervical cancer,cervical cancer tissue and normal tissue parameters were compared between the two groups.Results 35 patients were detected in the observation group,the detection rate was 87.5%,23 patients were detected in the control group,the detection rate was 57.5%,the detection rate of the observation group was significantly higher than the control group (x2=9.028,P=0.002).In the observation group,7 patients in stage Ⅰ(17.5%),1 case in the control group (2.5%),there was significant difference between the two groups (x2=5.000,P=0.025),the differences in phase Ⅱ,Ⅲ and Ⅳ were not statistically significant(P>0.05).In observation group,the peak intensity of cervical cancer was (65.43±10.55)%,and the peak time was (22.09±2.82)min,the peak intensity of normal tissue was (58.21±14.12)%,peak time was (28.27±3.01)min,the peak intensity of cervical cancer was higher than normal tissue,the peak time was shorter than normal tissues,the differences were statistically significant(t=2.590,9.476,all P0.05).Conclusion Compared with conventional ultrasonography,the contrast-enhanced ultrasonography can clearly show the range of tumor,quantitative analysis of time intensity curve parameters,can better reflect the characteristics of cervical cancer lesions,can improve the diagnostic rate and clear clinical stage.
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Objective To assess the left ventricular global systolic function changes using ultrasonic three -dimensional speckle tracking imaging (3 D -STI )in patients with uremia undergoing high -flux hemodialysis (HFHD)treatment.Methods 86 patients with uremia and 59 healthy volunteers were selected.Participants of control group underwent plasma brain natrinuretic peptide(BNP)examinations before 3D -STI examinations.All the uremia patients received HFHD therapy,and underwent BNP and 3D -STI examinations before and after 4 weeks HFHD therapy.Parameters of left ventricular three dimensional global area strain(GAS),global circumferential strain (GCS),global longitudinal strain(GLS)and global radial strain(GRS)were measured in each group.The correlation between each 3D -STI parameters and BNP level were analyzed.Results Compared with the control group,GAS, GCS,GLS and GRS of left ventricular were significantly lower(t =7.564,6.866,7.946,5.477,all P <0.05)and BNP level in the control group were significantly higher in the uremia group[(60.75 ±17.22)ng/L vs.(792.68 ± 184.56)ng/L,t =30.876,P <0.05].After HFHD therapy 4 weeks,GAS,GCS,GLS and GRS of left ventricular were significantly higher(t =4.233,5.933,6.546,2.959,all P <0.05)and BNP level were lower(t =27.533,P <0.05) than before treatment in the uremia group.GAS,GCS,GLS and GRS of left ventricular were negatively correlated with BNP level(r =-0.51,0.64,-0.46,-0.43,all P <0.05).Conclusion 3D -STI can accurately evaluate the changes of left ventricular global systolic function in patients with uremia before and after hemodialysis,and provide reliable basis for clinical treatment and efficacy assessment.
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AIM: To observe the effects of puerarin on blood lipid and expression of aorta laminin B_1 mRNA in streptozotocin-induced diabetic rats. METHODS: Diabetic nephropathy rats were induced by intraperitoneal injection of STZ, and the experimental rats were divided into normal control group, model group, and puerarin group. During and after the treatment for 12 weeks, the general state, blood suger(BS), triglyceride(TC), cholesterol, low density lipoprotein(LDL-C), high density lipoprotein(HDL-C), glycosylated hemoglobin(HbA1c) and glycosylated low density lipoprotein(G-LDL) were detected. Aorta alteration of tissue morphology was observed by H.E staining, and the expressions of laminin B1 mRNA were determined by in situ hybridization analysis. RESULTS: Diabetes mellitus and aorta lesion occurred in the two model groups. Puerarin could improve the general state, decrease the level of triglyceride(P
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Objective:To prepare polyclonal antibody against human immunoglobulin G(human IgG) and to use this antibody in the clinical diagnosis of nephrosis. Methods:New Zealand rabbit was immunized subcutaneously with human IgG.Enzyme linked immunosorbent assay(ELISA) was applied to reveal the titer of the prepared polyclonal antiserum against human IgG.Antiserum was purified with affinity chromatography,and the purified antibody was confirmed for its specificity by Western blot and immunohistochemical staining.The purified antbodies which have been indentified were fused with FITC(fluorescein isothiocyanate) and then analyzed by immunohistochemical staining.Finally the fused antibody was useed in the clinical diagnosis of nephrosis. Results: The titer of the obtained antiserum was up to(1∶128 000,) double agar diffusion test showed the titer of the antibody was 1∶32.By the method of affinity chromatography,we obtained purified antibody with the purity of 85%.ELISA,double agar diffusion and immunohistochemical staining tests showed that the specificity and titer of the antibody were not decreased sharply.The purified antibody fused with FITC also kept the specificity of the primitive antibody.When the FITC fused antibody was tried in nephrosis patients,it detected human IgG effectively. Conclusion:The polyclonal antibody can specifically recognize human IgG.This purified antibody fused with FITC can be used in the diagnosis of nephrosis.
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Aim To investigate the effects of bone morphogenic protein(BMP)-7 on the expression of extracelluar matrix components(ColⅠand FN)in human renal proximal tubular cells(HK-2)induced by transforming growth factor-?1(TGF-?1),and to explore the inhibition of BMP-7 on renal interstitial fibrosis.Methods HK-2 cells were treated with TGF-?1,BMP-7,or a combination of both for 48 h.The expression of FN was assessed by indirect immunofluorescence.RT-PCR and Western blot were used to determine the mRNA and protein expression of FN and ColⅠ?1.Results Indirect immunofluorescence staining showed that FN staining in 3 ?g?L-1 TGF-?1 group was considerably stronger than that in control group.By an addition of 200 ?g?L-1 BMP-7,the expression of FN was significantly inhibited.RT-PCR and Western blot showed the mRNA and protein expression of FN and ColⅠ?1 were up-regulated by TGF-?1 after 48 hours significantly(P
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ObjectiveTo investigate the influence of aging and castration on the density of andro gen receptor(AR) in the male rat heart. MethodsAR in the heart of rats aged 40d,90d(intact and castrated) and 2 years were demonstrated by means of immunohistochemical technique and the den sity of the positive staining was measured by using image analys is. The optical density of AR in the septum of the rats were analyzed by anova and doncan process of the SAS data analysis system. Re sultsOD(AR) of the group aged 2 years was higher than that of the groups aged 40d and 90d (0. 3550±0. 0077,0. 31080. 0264 and 0. 3149±0. 0200 respectively), thetwolattergroups showed similarly intensive staining, while the value of castrated group(0. 4136±0. 0320) was signif icantly greater than that of intact same aged group. Conclusion The development of sexual gland has little influence in the density of AR in the male rat heart during prepuberty to adult, both old age and castration increase the density of AR in the rat heart.
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AIM To explore the effects of sinomenine (Sin) on psychic dependence induced by morphine (Mor) in mice and its relation to the cAMP content of the cerebral cortex. METHODS A biased procedure induced the strong place preference. The effect of Sin (10, 30, 60 mg?kg -1 , ip) on the rewarding property of morphine was observed in a conditioned place preference(CPP) paradigm in mice,at the same time, the change of cAMP content in cerebral cortex was measured by the radioimmunoassay(RIA). RESULTS Sin significantly shortened the time of staying in morphine-pair compartment and lowered the cAMP level of mice. CONCLUSION Sin could attenuate effects of psychical dependence induced by Mor, its mechanism may be related to decrease of cAMP level in the brain, and Sin did not display psychical dependence in the experiment.
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Aim To investigate the potential suppressing effect of calmodulin antagonist EBB on metastasis-associated properties of human metastatic ovarian clear cell adenocarcinoma cell ES-2.Methods MTT assay was used to assess the growth inhibition of EBB on ES-2 cells.The invasive capacity and motility potential were determined by Transwell chamber assay and Wound assay.[Ca2+]i was observed under the laser scanning confocal microscopy.Results EBB inhibited the proliferation of ES-2 cells in vitro.The IC50 on ES-2 cells was(13.67?1.56)?mol?L-1.The invasive ability and motility potential of ES-2 cells were decreased after exposure to 3,7 and 14 ?mol?L-1 EBB respectively(P