ObjectiveTo clarify the graded quantitative diagnostic characteristics of lung qi deficiency syndrome in chronic obstructive pulmonary disease （COPD） based on latent class analysis combined with a hidden structure model. MethodsClinical data， including the four diagnostic methods of traditional Chinese medicine （TCM）， were collected from 745 COPD patients with lung qi deficiency syndrome. Latent class modeling was performed using R 4.1.2 software， and each patient was classified into one of three severity categories （mild， moderate， or severe） based on probabilistic parameterization， parameter estimation， and model fitting. A database was established for different severity levels of lung qi deficiency syndrome. Based on this， Lantern 5.0 software was used to construct hidden structure models for mild， moderate， and severe lung qi deficiency syndrome， and syndrome differentiation rules were developed through comprehensive clustering. ResultsA latent class model was constructed using 28 symptoms and signs with a frequency greater than 10%. Considering TCM theory and model simplicity， the optimal model was determined when the number of latent classes was three， categorizing lung qi deficiency syndrome into mild （298 cases）， moderate （164 cases）， and severe （283 cases）. Hidden structure models were separately developed for each severity level， and syndrome differentiation rules were established. A comparison of common symptoms in the syndrome differentiation rules for mild and moderate lung qi deficiency syndrome showed no statistically significant differences in diagnostic values and weights （P＞0.05）， leading to their combined analysis and the development of a unified syndrome differentiation rule. Value and weight of quantitative diagnosis of mild-to-moderate lung qi deficiency syndrome were as followed： shortness of breath （diagnostic value 9.3， diagnostic weight 86.92%）， dyspnea on exertion （8.2， 76.64%）， low voice and reluctance to speak （6.7， 62.62%）， poor appetite （4.0， 37.38%）， loose stools （4.0， 37.38%）， weak cough sound （2.9， 27.10%）， wheezing （2.3， 21.50%）， fatigue （1.8， 16.82%）， spontaneous sweating （1.7， 15.89%）， susceptibility to colds （1.6， 14.95%）， swollen tongue （1.4， 13.08%）， teeth marks on the tongue edge （1.2， 11.21%）， deep pulse （1.6， 14.95%）， with a diagnostic threshold of 10.3. Value and weight of quantitative diagnosis of severe lung qi deficiency syndrome were as followed： weak cough sound （15.1， 61.13%）， soreness and weakness of the waist and knees （12.6， 51.01%）， shortness of breath （11.1， 44.94%）， low voice and reluctance to speak （8.3， 33.60%）， frequent nocturia （6.1， 24.70%）， spontaneous sweating （3.7， 14.98%）， susceptibility to colds （3.5， 14.17%）， teeth marks on the tongue edge （7.8， 31.58%）， pale tongue body （1.9， 7.69%）， white tongue coating （5.5， 22.27%）， thin pulse （1.5， 6.07%）， with a diagnostic threshold of 23.7. ConclusionThe combination of latent class analysis and a hideen structure model effectively clarified the graded quantitative diagnostic characteristics of lung qi deficiency syndrome， providing a reference for the quantitative diagnosis of other fundamental syndromes in TCM.

Objective To assess the current state and influencing factors of multi-site work-related musculoskeletal disorders (WMSDs) among front-line employees in the assembly workshop of an automobile manufacturing enterprise. Methods A total of 394 front-line workers in the assembly workshop of an automobile manufacturing enterprise in Beijing City were selected as the research subjects using the judgmental sampling method. The Chinese version Musculoskeletal Disorders Questionnaire was used to assess the presence of WMSDs over the past 12 months in nine body regions: neck, shoulders, upper back, lower back, elbows, wrists, hips and thighs, knees, ankles and feet. The multivariable logistic regression was employed to investigate the influencing factors. Results The detection rate of overall WMSDs was 32.7% (129/394), with the top three single-site WMSDs being in the neck, shoulders, and lower back, and their detection rates were 14.0%, 12.7% and 9.6%, respectively. The detection rate of multi-site WMSDs was 17.8% (70/394). The result of multivariable logistic regression analysis revealed that workers who turned or bent their upper body while keeping their legs stationary, frequently performed wrist flexion/extension/lateral bending/rotation, or stood for prolonged period of time had significantly higher risks of developing multi-site WMSDs compared with those who did not (all P<0.05). Workers who perceived uncomfortable workplace lighting had higher risk of multi-site WMSDs than those who perceived it as comfortable (P<0.01). Conclusion The development of multi-site WMSDs among workers in the assembly workshop of this automobile manufacturing enterprise is strongly related to poor working postures at work.

Objective:To investigate the distribution characteristics and clinical significance of aeroallergen sensitization in children with airway allergic diseases.Methods:The information of children who were diagnosed with airway allergic diseases and performed with skin prick test(SPT)of aeroallergens was collected and retrospectively analyzed from Beijing Children′s Hospital from January to December, 2019.A total of 2557 patients were divided into ≤5 years of age group, 6-11 years of age group and ≥12 years of age group according to age, and allergic rhinitis group, asthma group, allergic rhinitis combined with asthma group according to diseases.The differences in the distribution of positive rates of 18 kinds of aeroallergens in age group and disease group were compared.Results:The positive rates of SPT in 2557 children from high to low were weeds pollens in summer and autumn, molds, trees pollens in spring, dust mites, etc..The top five strong positive rates from high to low were Artemisia pollen, Humulus pollen, Dermatophagoides farina, Betula pollen and Fraxinus pennsylvanica pollen.The positive rate was 71.5%(539/754)in the ≤5 years of age group, 78.5%(1241/1581)in 6-11 years of age group, and 81.1%(180/222)in ≥12 years of age group.The difference in positive rates of SPT among different age groups was statistically significant( χ2=16.825, P<0.05). In the group of ≤5 years of age group, the main aeroallergens were Chenopodium pollen(310/754, 41.1%), Alternaria alternate(307/754, 40.7%), Humulus pollen(295/754, 39.1%), Artemisia pollen(293/754, 38.9%)and Fraxinus pennsylvanica pollen(258/754, 34.2%). The main aeroallergens in the 6-11 years of age group were Chenopodium pollen(853/1581, 54.0%), Humulus pollen(769/1581, 48.6%), Artemisia pollen(768/1581, 48.6%), Alternaria alternate(751/1581, 47.5%)and Fraxinus pennsylvanica pollen(724/1581, 45.8%). The main aeroallergens in the ≥12 years of age group were Chenopodium pollen(131/222, 59.0%), Humulus pollen(119/222, 53.6%), Artemisia pollen(113/222, 50.9%), Alternaria alternate(112/222, 50.5%)and Dermatophagoides farina(103/222, 46.4%). The positive rate of allergic rhinitis group was 73.6%(1164/1582), asthma group was 72.4%(234/323), allergic rhinitis combined with asthma group was 86.0%(561/652). The positive rate of SPT was significantly different among different disease groups( χ2=43.408, P<0.05). The main aeroallergen of allergic rhinitis, asthma and allergic rhinitis combined with asthma was Chenopodium pollen.The positive rates of 18 kinds of aeroallergens were significantly different among the three disease groups(all P<0.05). The positive rate of each aeroallergen in allergic rhinitis combined with asthma was higher than that in single airway allergic disease. Conclusion:In children with airway allergic disease, the sensitization spectrum of aeroallergen varies among different age groups and disease groups.Clinicians should monitor allergens regularly and give preventive treatment to children with airway allergic disease.

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Objective To investigate the role of vitamin D in the synthesis and degradation of aggrecan in rat articular chondrocytes at cellular level. Methods Rat articular chondrocytes were stimulated by IL-1α, IL-1β and TNF-α, respectively. Normal and inflammatory chondrocytes were treated with different doses of vitamin D, respectively. CCK8, Flow cytometry, real time-PCR and western blot analysis were used to examine the proliferation activity and apoptosis level of chondrocytes, and the expression of aggrecan, ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels. Results IL-1α,IL-1β and TNF-α significantly decreased the proliferation activity and increased the apoptosis level of the chondrocytes. Furthermore, IL-1α, IL-1β and TNF-α significantly decreased the expression of aggrecan, and increased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes. 1α,25 (OH)2D3supplementation significantly increased the proliferation activity and decreased the apoptosis level of chondrocytes stimulated by IL-1α, IL-1β and TNF-α in a dose-dependent manner, but not affected the normal chondrocytes. Meanwhile, 1α,25(OH)2D3also significantly increased the expression of aggrecan, and decreased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes under inflammatory conditions. Conclusions Vitamin D may promote the anabolism of aggrecan and inhibit aggrecanase activity in chondrocytes under inflammatory conditions, which may impact overall protection for articular cartilage.

Objective To investigate the etiology composition of end-stage renal disease (ESRD) in children,in order to provide reference for the prevention and treatment.Methods The children with ESRD who were diagnosed in Peking University First Hospital from January 2005 to October 2013 were selected,and the etiology composition and incidence of the children with ESRD were retrospectively analyzed.Diagnostic criteria for children with ESRD refer to the clinical practice guidelines for chronic renal disease (NKF/KDOQI),developed by the American kidney foundation in 2002.Results Eighty-six children with ERSD were enrolled including 53 cases of males,33 cases of females,with the male to female ratio of 1.61 ∶ 1.00 and the mean onset age was (7.08 ± 4.23) years old,and their average diagnosis age was(9.25 ±4.17) years old.The median duration of ERSD before diagnosis was 0.84(0.01-13.67)years.The main cause of ESRD was acquired renal disease,accounting for 43.02％ (37/86 cases),mainly the chronic glomerulonephritis (18/86 cases,20.93％) and nephrotic syndrome (16/86 cases,18.60％);followed by urinary congenital abnormity,accounting for 40.70％ (35/86 cases),in which the most common were renal dysplasia (18/86 cases,20.93％) and cystic renal disease (11/86 cases,12.79％).Children under 3 years old mainly showed congenital urinary tract abnormalities(6/10 cases,60.00％).But children over 3 years old mainly showed acquired renal diseases (37/76 cases,48.7％),and pathologic classification of glomerular disease were proliferative mesangial glomerulonephritis (6/23 cases,26.09％),focal segmental glomerulosclerosis (5/23 cases,21.74％) and interstitial nephritis(3/23 cases,13.04％).Conclusions The main etiology of ESRD is glomerular disease and congenital abnormal development of urinary system,therefore,more attention should be paid on the ultrasound screening of the urinary tract in the perinatal period and urine screening in children.There are great significances in reducing the incidence of ESRD and intervening actively the progression to chronic kidney disease.

Objective To investigate the levels and significance of Th17 cells and regulatory T cells (Treg) in peripheral blood of children with allergic rhinitis during pollen and non-pollen seasons.Methods Thirteen children with hay fever, 10 children with house dust mite(HDM)-allergic asthma and 10 healthy children were recruited into this study.Percentages of Th17 and Treg cells were detected by flow cytometry.Levels of IL-17, IL-10 and TGF-β in cell culture supernatants were measured by ELISA.Results (1) The percentages of Th17 cells in children with allergic rhinitis [(3.4±2.4)%] were significantly higher than those in HDM-allergic asthmatics [(2.1±1.6)%] and those in healthy children [(0.5±0.3)%] during pollen season (both P<0.05).The levels of Treg cells in allergic rhinitis group [(2.1±1.3)%] and in HDM-allergic asthma group [(3.6±1.9)%] were significantly lower than those in healthy control group [(5.5±2.8)%] (both P<0.05).The levels of Th17 cells [(3.0±1.9)% vs (3.4±2.4)%, P<0.05] and ratios of Th17/Treg cells [(1.4±1.0)% vs (1.7±1.5)%, P<0.05] in children with allergic rhinitis were significantly decreased during non-pollen season as compared with those during pollen season, but the levels of Treg cells were up-regulated [(2.4±1.6)% vs (2.1±1.3)%, P<0.05].(2) Correlation analysis revealed that the ratios of Th17/Treg cells were positively correlated with the concentrations of FeNO (fractional concentration of exhaled NO) (r=0.321, P<0.05) and the counts of circulating eosinophils (r=0.198, P<0.05) in children with allergic rhinitis during pollen season.Conclusion The imbalanced Th17 and Treg cells in children with allergic rhinitis during pollen season might play a vital role in the regulation of allergic airway inflammation.

Objective To investigate the changes in percentage and function of CD4+CD25+regu-latory T cells ( Tregs) in peripheral blood of patients with hay fever. Methods A total of 20 patients with hay fever, 20 patients with house dust mite-induced allergic asthma and 20 healthy subjects were enrolled in this study. Peripheral blood samples were collected from all subjects to isolate PBMCs. Percentages of Tregs in PBMCs were measured by flow cytometry. CD4+CD25+ Tregs and CD4+CD25-T cells ( Teffs) were isola-ted by immunomagnetic cell sorting. Effects of CD4+CD25+Tregs on the proliferation of Teffs were evaluated by MTT assay. Expression of Foxp3 and TGF-β1 at mRNA level was analyzed by RT-PCR. Results During the pollen season, the percentage of circulating Tregs in patients with hay fever [(1. 82+0. 82)％] was sig-nificantly lower than that in patients with house dust mite-induced allergic asthma [(2. 96±1. 34)％] and health subjects [(5. 78±2. 29)％] (both P<0. 05). Expression of Foxp3 at mRNA level was significantly reduced in patients with hay fever (0. 46±0. 25) as compared with that of the house dust mite-induced aller-gic asthma (0. 64±0. 31) and healthy control (1. 04±0. 21) groups (both P<0. 05). Expression of TGF-β1 at mRNA level in both hay fever (0. 34±0. 27) and house dust mite-induced allergic asthma (0. 43±0. 31) groups was lower than that of the healthy control group (0. 99±0. 34). Treg-mediated suppression of Teff proliferation was significantly decreased in patients with hay fever [(17. 1±8. 4)％] as compared with that in patients with house dust mite-induced allergic asthma [(21. 4±9. 1)％]) and healthy subjects [(36. 0± 13. 9)％] (P<0. 05). Conclusion Decreased percentage and defective function of Tregs might be one of the major causes for the occurrence and development of hay fever in children during the pollen season.

OBJECTIVETo analyze the clinical and genetic features of X-linked Alport syndrome (XLAS) in men positive for the collagen α5(Ⅳ) chain in epidermal basement membrane.

METHODThis was a retrospective study. Totally 725 families were diagnosed as Alport syndrome in Department of Pediatrics of Peking University First Hospital during January 1998 to December 2014, among them 450 patients were males with XLAS. Patients who met both of the following two criteria were included in this study. (1)Patients underwent α5(Ⅳ) chain staining in the epidermal basement membrane. (2)Mutations in COL4A5 gene were detected.Mann-Whitney test and χ(2) test were used.

RESULTTotally 140 males with XLAS were included in this study, 18 cases were α5 (Ⅳ)-positive and 122 cases were α5 (Ⅳ)-negative. The two groups of patients were compared, the median age at analysis was 11.0 vs. 7.2 years (Z = -1.839, P = 0.066), the 24-hour urine protein was 1.50 vs. 0.57 g/d (Z = -1.212, P = 0.226), the rate of hearing loss was 28% vs. 53% (χ(2) = 3.619, P = 0.067), the number of patients progressed to end stage renal disease (ESRD) was 4 vs. 12 (χ(2) =2.377, P = 0.128), the median age of ESRD was 31.0 vs. 16.6 years (Z = -2.554, P = 0.011), the rate of missense mutations in COL4A5 gene was 67% vs. 52% (χ(2) = 1.424, P = 0.313).

CONCLUSIONCompared the two groups of patients with positive and negative staining for the collagen Ⅳ α5 chain in epidermal basement membrane, there was no significant difference in the proteinuria level, the rate of hearing loss and genotype of COL4A5 gene. But the patients with positive staining progressed to ESRD significantly later than the patients with negative staining.

Basement Membrane ; pathology ; Child ; Collagen Type IV ; genetics ; DNA Mutational Analysis ; Deafness ; Humans ; Kidney Failure, Chronic ; Male ; Mutation, Missense ; Nephritis, Hereditary ; genetics ; pathology ; Proteinuria ; Retrospective Studies