This study analyzed the outcome index and related design elements of randomized controlled trials （RCTs） of traditional Chinese medicine （TCM） in the treatment of children with tic disorder （TD） in the past ten years， so as to provide a basis for the construction of the core index set of TCM in the treatment of children with TD. Eight databases were searched， including four English databases （PubMed， Web of Science， Embase， and Cochrane Library） and four Chinese databases， including China National Knowledge Infrastructure （CNKI）， Wanfang Database， VIP Database， and China Biology Medicine disc （CBMdisc）， as well as ClinicalTrials.gov and China Clinical Trial Registry. The search time was limited to from January 1， 2013 to October 29， 2023. RCTs on the TD in children treated with TCM were collected. Two researchers independently conducted literature screening， data extraction， and literature quality evaluation and summarized clinical outcome indexes and related trial design elements through qualitative analysis. A total of 67 RCTs were included， including 63 outcome indexes， with a total frequency of 348 times. The related outcome indexes could be divided into six categories： 12 symptom/sign indexes with a frequency of 134 （38.5%）， seven TCM symptom/syndrome indexes with a frequency of 31 （8.9%）， 33 physical and chemical examination indexes with a frequency of 97 （27.9%）， four safety indexes with a frequency of 67 （19.3%）， three long-term prognostic indexes with a frequency of 14 （4.0%）， and one kind of quality-of-life evaluation index （0.3%）. Currently， the RCTs research design of TCM in the treatment of TD in children has not yet formed a unified standard， and there are many problems in the quality of methodology， which reduces the authenticity and reliability of clinical conclusions. There are problems with clinical outcome indexes， such as significant quantity differences， unclear primary and secondary outcome indexes， unreasonable alternative indexes， non-standard TCM syndrome types and TCM evaluation indexes， lack of economic evaluation indexes， and less attention to long-term prognostic indexes and safety indexes. It is suggested that the researchers should design a more rigorous trial scheme and reasonably design the outcome index which is in line with the clinical trial efficacy evaluation of TCM， so as to construct the core index set with the characteristics of TCM for the treatment TD in children.

To summarize the clinical experience of Professor YAN Huimin in pattern identification and treatment for chest tightness variant asthma in children with the method of regulating qi movement. It is believed that children's chest tightness variant asthma is mainly located in lungs and involves liver and spleen， and the core mechanism of the disease is disturbance of qi movement. On the basis of regulating qi， syndrome differentiation and treatment is conducted： for pattern of lung qi deficiency and cold， phlegm-fluid retention， the treatment is appropriate to tonify the lung and benefit qi， and warm phlegm-fluid， which commonly used in modified Yupingfeng Powder （玉屏风散） and Xiaoqinglong Decoction （小青龙汤）； for pattern of phlegm and qi binding constraint， the treatment is appropriate to soothe the liver and resolve constraint， and dissolve phlegm and dissipate masses， which commonly used in modified Banxia Houpo Decoction （半夏厚朴汤） and Jinlingzi Powder （金铃子散）； for pattern of qi deficiency and blood stasis， the treatment is appropriate to tonify the deficiency to reinforce healthy qi， and move qi to invigorate blood， which commonly used in modified Xuefu Zhuyu Decoction （血府逐瘀汤）. It is emphasised that during the treatment process， the developmental dynamics of the disease should be grasped， patterns and treat should be identified， and special attention to the changes of qi movement should be paid.

Humans ; Asthma/drug therapy* ; Biological Therapy

The aim of this study was to construct a recombinant adenovirus expressing extracellular domain gene of human epidermal growth factor receptor variant Ⅲ (EGFRvIII ECD), and to prepare single domain antibody targeting EGFRvIII ECD by immunizing camels and constructing phage display antibody library. Total RNA was extracted from human prostate cancer cell line PC-3 cells and reversely transcribed into cDNA. EGFRvIII ECD gene was amplified using cDNA as template, and ligated into pAdTrack-CMV plasmid vector and transformed into E. coli BJ5183 competent cells containing pAdEasy-1 plasmid for homologous recombination. The recombinant adenovirus expressing EGFRvIII ECD was obtained through transfecting the plasmid into HEK293A cells. The recombinant adenovirus was used to immunize Bactrian camel to construct EGFRvIII ECD specific single domain antibody library. The single domain antibody was obtained by screening the library with EGFRvIII protein and the antibody was expressed, purified and identified. The results showed that recombinant adenovirus expressing EGFRvIII ECD was obtained. The capacity of EGFRvIII specific phage single domain antibody library was 1.4×10⁹. After three rounds of enrichment and screening, thirty-one positive clones binding to EGFRvIII ECD were obtained by phage-ELISA, and the recombinant single domain antibody E14 with highest OD₄₅₀ value was expressed and purified. The recombinant E14 antibody can react with EGFRvIII ECD with high affinity in ELISA assessment. The results indicated that the EGFRvIII specific single domain antibody library with high capacity and diversity was constructed and the single domain antibody with binding activity to EGFRvIII was obtained by screening the library. This study may facilitate the diagnosis and treatment of EGFRvIII targeted malignant tumors in the future.
Adenoviridae/genetics* ; DNA, Complementary ; ErbB Receptors ; Escherichia coli/genetics* ; Genetic Vectors/genetics* ; Humans ; RNA ; Recombinant Proteins/metabolism* ; Single-Domain Antibodies

Objective:To compare the effects of different intraocular infusion solutions on histology and function of retina.Methods:Human corneal endothelial cells (HCEC), human retinal pigment epithelium (HRPE) cells and rat retinal ganglion cells (RGC) were divided into normal control group, balanced saline solution (BSS) group and compound electrolyte intraocular irrigating solution (CEIIS) group, and the cells were cultured in 10% DMEM/F12 medium, BSS and CEIIS for 12, 24 and 48 hours, respectively, according to grouping.The proliferation absorbance value of cultured cells was measured by cell counting kit-8 (CCK8) method.The expression of apoptosis related proteins in cultured cells was detected by cellular immunofluorescence staining.The cell apoptosis rate and cell cycle were measured by flow cytometry.The mitochondrial damage was detected by lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) quantitative detection kit.Fifteen New Zealand white rabbits were randomly divided into control group ( n=3), BSS group ( n=6) and CEIIS group ( n=6). The left eyes were taken for vitrectomy and different intraocular perfusion fluids were used during vitrectomy according to grouping.The retinal function of operative eyes was measured by flash electroretinogram (ERG) before operation and 24 hours after operation, and the structural changes of each layer of retina were detected by optical coherence tomography (OCT). The early apoptosis of retinal cells was detected by TUNEL staining.The expressions of cytochrome C and bax protein in retina were detected by immunohistochemical staining.The ultrastructural changes of retina were observed under a transmission electron microscope.The use and care of animals complied with the ARVO statement.This study protocol was approved by an Ethics Committee of Peking University People's Hospital (No.2019PHE059). Results:The three kinds of cultured cells in BSS and CEIIS groups were damaged in various degrees.With the extension of culture time, proliferated cells were decreased and the number of apoptotic cells was increased.Compared with the BSS group, cultured cells in the CEIIS group were dense and in orderly arrangement with uniform morphology and size.The apoptosis rates of HRPE cells and RGC in the BSS group were (37.157±6.918)% and (29.993±12.330)%, respectively, which were significantly higher than (4.163±1.310)% and (6.337±1.903)% in the CEIIS group ( P=0.003, 0.045). There was no significant difference in G0/G1+ S phase ratio of HCEC and HRPE cells among the normal control group, BSS group and CEIIS group (HCEC: F=2.226, P=0.189; HRPE: F=2.634, P=0.151), and the proportion of G2/M division arrest phase of RGC in the BSS group was significantly higher than that in the normal control group and CEIIS group ( P=0.047, 0.024). The proliferation absorbance values of HCEC, HRPE cells and RGC in the CEIIS group were significantly higher than those in the BSS group at each culture time point (all at P<0.05). The fluorescence intensity of cytochrome C, bax, caspase-3 and caspase-9 proteins in the BSS group was stronger than that in the normal control group and CEIIS group, and the fluorescence intensity of bcl-2 was weaker than that in the CEIIS group, and the fluorescence intensity of zonula occluden-1 (ZO-1) was weaker than that in the normal control group and CEIIS group.The release level of LDH in the BSS group was significantly higher than that in the CEIIS group at different time points (all at P<0.001). After 48 hours of culture, the release level of SDH in the BSS group was significantly higher than that in the CEIIS group ( P<0.05). No retinal histological abnormalities was found through OCT examination of rabbit eyes after vitrectomy in the two groups, but transmission electron microscopy showed that there were different degrees of loose arrangement of retinal photoreceptor cells, a large number of photoreceptor outer membrane discs falling off and vacuolar degeneration in the two groups, especially in the BSS group.TUNEL staining showed that the apoptotic cells were mainly located in the inner nuclear layer and RGC layer.The number of apoptotic retinal cells was (135.2±22.8)/high-power field of vision in the BSS group, which was significantly higher than (81.3±17.7)/high-power field of vision in the CEIIS group ( t=4.175, P=0.002). Full field flash ERG showed that the amplitudes of scotopic 3.0 ERG a- and b-wave in the CEIIS group after operation were significantly lower than those before operation, but the differences were not statistically significant (all at P>0.05). The amplitudes of scotopic 3.0 ERG a- and b-wave in the BSS group after operation were significantly lower than those before operation ( P=0.026, 0.010). Conclusions:In vivo and in vitro research results show that compared with BSS, there were few apoptotic cells in retinal tissue after vitrectomy perfused by CEIIS.

Objective: To explore the effectiveness and safety of ultrasound-guided radiofrequency ablation (RFA) in treatment of thyroid benign nodules. Methods: We analyze 573 patients with thyroid benign nodules from June 2014 to September 2017 treated by RFA at Department Ⅱof Thyroid Surgery, the First Affiliated Hospital of Zhengzhou University. Among these patients, there were 75 males and 498 females, with a median age of 45 years old. All patients were diagnosed as thyroid benign nodules by ultrasound-guided fine needle aspiration biopsy before RFA. A total of 750 benign tumors were treated. To evaluate the thyroid function of the patients before RFA and 3 months after it, and to observe the changes of thyroid benign nodules by ultrasound at 3, 6, 12 months after RFA. The paired t-test was used to compare the measurement data with normal distribution, and Wilcoxon's signed rank test was used to compare the measurement data with non-normal distribution. To calculate the volume change and reduction rate of thyroid benign nodules. Results: RFA was successfully completed in all patients, the volume reduction rate was 67%(48%, 83%) in the 3rd month after RFA, in the 6th month was 81%(67%, 91%), in the 12th month was 89%(80%, 95%). Eighteen patients felt pain during RFA, but the pain was alleviated after stopping ablation. Three patients′ tone decreased, but recovered in a week. Hoarseness occurred in 6 patients and recovered in 3 months. Three patients had neck hemorrhage, which was managed with simple compression of the neck. Conclusions RFA is an effective and safe treatment for thyroid benign nodules and has obvious advantages such as less invasiveness, having no influence in thyroid functions. It is clinically prospective for application.

Objective:To investigate expression profiles of the plasma exosomal miRNAs of the chronic hepatitis B (CHB) patients with persistently normal alamine aminotransferase (PNALT) for the first time and try to find exosomal miRNAs which could reflect liver inflammation better.Methods:Five CHB patients with liver tissue inflammation grade ≥A2 of PNALT and 5 CHB patients with liver tissue inflammation grade ＜A2 of PNALT were enrolled and their blood samples were collected.The exosomes were extracted from these blood samples and measured by electron microscope to determine the extraction effect.The exosomal miRNAs were extracted and sent for high throughput sequencing,and the expression of exosomal miRNAs in the 2 groups of patients was analyzed.Results:Under the electron microscope,exosomes were small membranous vesicles with 30-100 nm in diameter.The peak value of particle size ranged from 10 to 100 nm.High throughput sequencing showed that there were 591 differentially expressed exosomal miRNAs between the 2 groups.Compared with the control group,18 exosomal miRNAs were up-regulated and 6 exosomal miRNAs were down-regulated in PNALT patients with the liver tissue inflammation grade ≥ A2.Conclusion:Exosomal miRNAs in the CHB patients with PNALT who have the different grades of liver inflammation are differently expressed.Some of the differently expressed exosomal miRNAs are expected to be sensitive biomarkers for timely assessment of liver inflammation in the CHB patients with PNALT.

Objective To investigate the substantia nigra (SN) and brainstem raphe (BR) echogenic features of Parkinson's disease (PD) patients with musculoskeletal pain.Methods A total of 115 PD patients recruited in the Second Affiliated Hospital of Soochow University from October 2014 to May 2016 were assessed with the following rating scales:Unified Parkinson's Disease Rating Scale (UPDRS),Hoehn and Yahr Staging Scale (H/Y),Hamilton Rating Scale for Depression (HRSD),Beck Depression Inventory Ⅱ (BDI-Ⅱ) and Visual Analogue Scale (VAS).All the subjects underwent transcranial sonography during the clinical evaluation.And the patients were divided into PD with musculoskeletal pain (n =54) and PD without musculoskeletal pain (n =61) groups,or PD with depression(n =74) and PD without depression(n =41) groups.Results Compared with PD patients without pain,PD patients with musculoskeletal pain had higher scores of UPDRS-Ⅱ,-Ⅲ,HRSD,BDI,NMSQ and H/Y (UPDRS-Ⅱ score:12.56 ±6.01 vs 8.79 ±4.38,t =-3.801,P ＜0.01;UPDRS-Ⅲ score:24.43 ± 12.43 vs 20.07 ± 11.12,t=-1.986,P=0.049;HRSD score:11.65-±6.94 vs 8.38-±5.36,t=-2.844,P=0.005;BDI score:14.09 ±6.20 vs 9.74 ±6.00,t =-3.826,P ＜0.01;NMSQ score:8.57 ± 4.06 vs 5.60 ± 3.38,t=4.193,P＜0.01;H/Y:2.0(1.5,2.6) vs 1.5(1.0,2.0),Z=-3.011,P=0.003).Positive BR was more frequent in depressed than in non-depressed PD patients without pain (63.6％ vs 14.3％;x2 =15.25,P ＜0.01).Positive BR was positively associated with sex(r =0.228,P =0.014),age(r =0.184,P =0.049),disease duration (r =0.196,P =0.035),and depression (r =0.396,P ＜ 0.01).However,positive BR did not correlate with musculoskeletal pain.No correlation was found between positive SN and clinical characteristics of PD patients.Conclusions PD patients with musculoskeletal pain have worse activity of daily living,more severe motor symptoms,more non-motor symptoms,and are more depressed.SN and BR echogenecity do not correlate with musculoskeletal pain,however,hypoechogenic or interrupted BR is associated with depression in PD patients.

OBJECTIVETo investigate the influence of maternal atopy on cord blood effector T cells and to identify these biologic markers as predictors of atopic dermatitis (AD).

METHODSSeventy mother-infant pairs were recruited in this prospective birth cohort study. Suspected factors for allergy, including maternal allergic history, total serum IgE, and maternal age at birth, were collected. Mother peripheral blood samples and cord blood were obtained and assayed for the percentage of interferon-γ (IFN-γ) and interleukin 4 (IL-4) producing T cells(Th1 and Th2 respectively) using flow cytometry. Their offspring at the age of 2 years old were evaluated by their dermatologist whether they had AD. Statistical analysis was performed using multiple logistic regression models and receiver-operating characteristic curve was employed to predict atopic dermatitis.

RESULTSTwenty-one allergic and 49 nonallergic mothers were recruited in this study. During the first two years of life, 15.7% children (n = 11) developed a physician-diagnosed AD (all children were the only child in the family). In group with maternal allergic rhinitis, a significantly increased percentage of Th2 was observed in peripheral blood of mother (7.10[1.18;16.1]% vs. 0.37[0.25;0.72]%, U = 10.0, P < 0.05) and cord blood of newborns (1.02[0.57;1.34]% vs. 0.21[0.15;0.42]%, U = 127.5, P < 0.05), respectively. Maternal atopic history did not affect the percentage of Th1 cells in cord blood (0.69[0.40;1.12]% vs.0.50[0.31;0.66]%, U = 361.0, P > 0.05). Children with reduced Th1/Th2 ratio in cord blood had a higher risk to develop AD (OR = 1.72, P = 0.001) . The model including Th1/Th2, maternal allergy, maternal age at birth and maternal total IgE showed high ability to discriminate children with and without AD. AUC was 0.907 (95% CI: 0.804-1.011, P < 0.001).

CONCLUSIONSElevated IL-4⁺CD4⁺ T cells in cord blood were of relevance with maternal allergic history. Imbalance between Th1 cell and Th2 cell at birth are associated with maternal allergy and promoted subsequent AD development.

Adult ; Child, Preschool ; Dermatitis, Atopic ; immunology ; Female ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Immunoglobulin E ; blood ; Infant ; Infant, Newborn ; Mothers ; Prospective Studies ; Rhinitis, Allergic ; blood ; immunology ; Th1 Cells ; cytology ; Th1-Th2 Balance ; Th2 Cells ; cytology

Objective To investigate the influence of maternal atopy on cord blood effector T cells and to identify these biologic markers as predictors of atopic dermatitis ( AD).Methods Seventy mother-infant pairs were recruited in this prospective birth cohort study.Suspected factors for allergy , including maternal allergic history , total serum IgE , and maternal age at birth , were collected.Mother peripheral blood samples and cord blood were obtained and assayed for the percentage of interferon -γ( IFN-γ) and interlukin 4 (IL-4) producing T cells(Th1 and Th2 respectively) using flow cytometry.Their offspring at the age of 2 years old were evaluated by their dermatologist whether they had AD.Statistical analysis was performed using multiple logistic regression models and receiver-operating characteristic curve was employed to predict atopic dermatitis.Results Twenty-one allergic and 49 nonallergic mothers were recruited in this study.During the first two years of life, 15.7%children (n=11) developed a physician-diagnosed AD (all children were the only child in the family ).In group with maternal allergic rhinitis , a significantly increased percentage of Th2 was observed in peripheral blood of mother ( 7.10 [ 1.18;16.1 ]% vs.0.37 [ 0.25;0.72]%, U=10.0, P <0.05 ) and cord blood of newborns ( 1.02 [ 0.57;1.34 ]% vs.0.21 [ 0.15;0.42]%, U=127.5, P<0.05), respectively.Maternal atopic history did not affect the percentage of Th1cells in cord blood (0.69[0.40;1.12]% vs.0.50[0.31;0.66]%, U=361.0, P>0.05).Children with reduced Th1/Th2 ratio in cord blood had a higher risk to develop AD (OR=1.72,P=0.001).The model including Th1/Th2, maternal allergy, maternal age at birth and maternal total IgE showed high ability to discriminate children with and without AD.AUC was 0.907 (95% CI:0.804 -1.011, P<0.001).Conclusions Elevated IL-4 +CD4 +T cells in cord blood were of relevance with maternal allergic history.Imbalance between Th1 cell and Th2 cell at birth are associated with maternal allergy and promoted subsequent AD development.