OBJECTIVE To investigate the ameliorating effect and mechanism of the effective substance groups from Artemisia ordosica（Abbreviated as HSH） on rheumatoid arthritis （RA） based on cluster of differentiation 4/lymphocyte cell-specific protein- tyrosine kinase/zeta-chain-associated protein kinase of 70 kDa/interleukin-17（CD4/LCK/ZAP70/IL-17） signaling pathway. METHODS The rats were divided into normal group， model group， HSH low-dose， medium-dose and high-dose groups （2.7， 10.8， 21.6 mg/kg） and positive control group （Tripterygium glycosides tablet， 6.3 mg/kg）， with 10 rats in each group. Except for normal group， RA rat model was induced by complete Freund’s adjuvant in other groups. After modeling， each group was given relevant medicine or normal saline intragastrically， once a day， for 28 consecutive days. The changes in ankle joint swelling and arthritis index in rats were determined； the pathological changes of ankle joint tissue were observed； the levels of inflammatory factors [IL-1β， IL-21， IL-17A， IL-2， interferon-γ （IFN-γ） and IL-6] in serum and joint fluid of rats were determined； the levels of Th1， Th17 and Treg cells in the whole blood and spleen of rats were detected； protein and mRNA expressions of LCK， proto-oncogene tyrosine-protein kinase Fyn （Fyn）， ZAP70， CD45， RAR-related orphan receptor γt （RORγt）， and forkhead box protein 3 （Foxp3） in ankle synovial tissue were determined. RESULTS Compared with normal group， the changes in ankle joint swelling， arthritis index， the levels of IL-1β， IL-21， IL-17A， IL-2， IFN-γ and IL-6 in serum and joint fluid， the levels of Th1， Th17 cells and Th17/Treg value in whole blood and spleen， and the protein and mRNA expression levels of LCK， Fyn， ZAP70， CD45， and RORγt in ankle joint synovium were all significantly increased/elevated （P＜0.05）. The level of Treg cells in the spleen， as well as the protein and mRNA expression levels of Foxp3 in the ankle joint synovium were significantly decreased （P＜0.05）. Compared with model group， most of the above-mentioned indicators were significantly reversed in the positive control group and all dose groups of HSH （P＜0.05）. The degree of pathological changes in ankle joint tissues was markedly improved， and inflammation was alleviated. CONCLUSIONS HSH can regulate the cascade reactions in the CD4/LCK/ZAP70/IL-17 pathway within the T-cell receptor signaling pathway， thereby modulating the Th17/Treg balance. This leads to the suppression of inflammatory responses and the alleviation of synovial tissue damage in rats with RA.

ObjectiveTo establish a geographical origin identification model for Foeniculi Fructus from Haiyuan， providing a new technical reference for the protection of Haiyuan's geo-authentic medicinal materials and its designation as a national geographical indication agricultural product. MethodsSamples of Foeniculi Fructus were collected from eight producing areas， including Minqin （Gansu）， Bozhou （Anhui）， Qingdao （Shandong）， Dezhou （Shandong）， Urumqi （Xinjiang）， Nujiang （Yunnan）， Gutuo （Inner Mongolia）， and Haiyuan （Ningxia）. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to detect the volatile organic compounds （VOCs） in samples from these geographic origins. VOCs were qualitatively analyzed through dual matching with the National Institute of Standards and Technology （NIST） mass spectral database and the IMS drift time database. Using the Reporter module and Gallery Plot visualization tools within the LAV analytical platform， VOC fingerprint profiles characterizing geographic origins were constructed. A non-targeted analytical strategy was adopted， and 97 VOCs detected via GC-IMS were subjected to principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA） based on their differential distribution patterns to construct an origin identification model for Foeniculi Fructus from Haiyuan region. Key discriminative markers were screened using variable importance in projection （VIP） values greater than 1. ResultsA total of 97 VOCs were identified， including alcohols， aldehydes， ketones， esters， organic acids， terpenoids， ethers， alkenes， and benzenes. The PLS-DA model， based on VOCs data obtained by GC-IMS， effectively distinguished Foeniculi Fructus in Haiyuan region from those of other origins. During cross-validation， the model achieved a prediction parameter （Q²） of 0.976 and a goodness-of-fit parameter （R²） of 0.936， with no overfitting observed in permutation testing. Twelve key flavor markers with VIP > 1 were identified as characteristic indicators of Haiyuan origin. ConclusionA stable and highly predictive origin identification model for Foeniculi Fructus from Haiyuan was successfully established using GC-IMS technology， PLS-DA， and VIP-based marker screening. This model provides a novel technical strategy for accurately distinguishing Foeniculi Fructus in Haiyuan region from other regional varieties and offers new technical support for its protection as a geo-authentic medicinal material and a nationally designated geographical indication agricultural product in China.

ObjectiveTo establish a geographical origin identification model for Foeniculi Fructus from Haiyuan， providing a new technical reference for the protection of Haiyuan's geo-authentic medicinal materials and its designation as a national geographical indication agricultural product. MethodsSamples of Foeniculi Fructus were collected from eight producing areas， including Minqin （Gansu）， Bozhou （Anhui）， Qingdao （Shandong）， Dezhou （Shandong）， Urumqi （Xinjiang）， Nujiang （Yunnan）， Gutuo （Inner Mongolia）， and Haiyuan （Ningxia）. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to detect the volatile organic compounds （VOCs） in samples from these geographic origins. VOCs were qualitatively analyzed through dual matching with the National Institute of Standards and Technology （NIST） mass spectral database and the IMS drift time database. Using the Reporter module and Gallery Plot visualization tools within the LAV analytical platform， VOC fingerprint profiles characterizing geographic origins were constructed. A non-targeted analytical strategy was adopted， and 97 VOCs detected via GC-IMS were subjected to principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA） based on their differential distribution patterns to construct an origin identification model for Foeniculi Fructus from Haiyuan region. Key discriminative markers were screened using variable importance in projection （VIP） values greater than 1. ResultsA total of 97 VOCs were identified， including alcohols， aldehydes， ketones， esters， organic acids， terpenoids， ethers， alkenes， and benzenes. The PLS-DA model， based on VOCs data obtained by GC-IMS， effectively distinguished Foeniculi Fructus in Haiyuan region from those of other origins. During cross-validation， the model achieved a prediction parameter （Q²） of 0.976 and a goodness-of-fit parameter （R²） of 0.936， with no overfitting observed in permutation testing. Twelve key flavor markers with VIP > 1 were identified as characteristic indicators of Haiyuan origin. ConclusionA stable and highly predictive origin identification model for Foeniculi Fructus from Haiyuan was successfully established using GC-IMS technology， PLS-DA， and VIP-based marker screening. This model provides a novel technical strategy for accurately distinguishing Foeniculi Fructus in Haiyuan region from other regional varieties and offers new technical support for its protection as a geo-authentic medicinal material and a nationally designated geographical indication agricultural product in China.

Objective To precisely identify the para-Bombay blood types in cancer patients at our hospital, establish a robust system for the identification of challenging blood types in our laboratory, and provide a foundation for precise transfusion practices. Methods We retrospectively analyzed the blood type results of 91 874 cancer patients from January 1, 2019, to December 31, 2023. Conventional serological methods were used to screen for blood types, and suspected para-Bombay blood types were identified. Further analysis was performed using Pacific Biosciences (PacBio) single-molecule real-time sequencing and Sanger sequencing was used to determine the genotypes of the ABO, FUT1, and FUT2 genes. Results Eight cases of para-Bombay blood type were confirmed through serological and molecular biological methods. The FUT1 genotypes identified were: 5 cases of h1h1 (homozygous mutation 551_552delAG) and 3 cases of h1h2 (compound heterozygous mutations of 551_552delAG and 880_882delTT). The FUT2 genotypes identified were: 2 cases of Se³⁵⁷/Se^{357, 716} and 4 cases of Se³⁵⁷/Se³⁵⁷. Additionally, one sample revealed a novel heterozygous mutation, 818C>T, in exon 7 of the ABO gene, which was confirmed by PacBio sequencing to be located on the O haplotype. Conclusion PacBio sequencing technology demonstrates significant advantages in analyzing the haplotypes of para-Bombay blood type genes. This approach supports the establishment of a robust system for the identification of challenging blood types and provides novel evidence for precise transfusion practices in cancer patients.
Humans ; Neoplasms/genetics* ; Fucosyltransferases/genetics* ; ABO Blood-Group System/genetics* ; Male ; High-Throughput Nucleotide Sequencing/methods* ; Galactoside 2-alpha-L-fucosyltransferase ; Female ; Retrospective Studies ; Genotype ; Middle Aged ; Blood Grouping and Crossmatching/methods* ; Adult ; Mutation ; Aged

Objective To explore the effects of HPV16 E6 on genes and signaling pathways in cervical epithelial cells and to screen genes associated with oncogenic transformation.Methods HUCEC models infected with HPV16 E6 were constructed,and transcriptome sequencing was performed to screen for differentially expressed genes(DEGs),which were subjected to Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment to analyze the differential signaling pathways.RT-qPCR was used to validate major differentially down-regulated expressed genes.After predicting the major differentially expressed proteins by molecular docking analysis,the expression of major differential genes in HUCEC cell model was verified by RT-qPCR and Western blotting.In addition,RT-qPCR and Western blotting were used to further verify the expression of major differential genes in cervical cancer cell lines,SiHa and CaSki.Results A total of 55 genes with more than two-fold differential expression were screened.The results centering on down-regulated genes showed that the negatively regulated differential gene was mainly enriched in redox processes;KEGG enrichment analysis revealed that it was mainly associated with carbohydrate metabolism and cancer.RT-qPCR results showed that the down-regulated differential expression trends of the selected 10 genes were basically consistent with the sequencing results.Molecular docking analysis predicted an interaction between DHRS2 and HPV16 E6,and RT-qPCR and Western blotting confirmed that HPV16 E6 down-regulated DHRS2 mRNA(P＜0.01)and protein(P＜0.05)and ETV5 protein expression(P＜0.01).In SiHa and CaSki cells,compared with the control group,the mRNA and protein expression of DHRS2 was downregulated and positively correlated with the trend of P53 protein expression(P＜0.05).Conclusion HPV16 E6 can mediate oncogenic transformation of cervical cells and promote cervical carcinogenesis through downregulating DHRS2 expression.

BACKGROUND:Rheumatoid arthritis is a chronic autoimmune disease characterized by joint pain,swelling,and dysfunction.Acupuncture,as a traditional medical treatment,has proved its effectiveness and safety in many diseases.However,the efficacy of acupuncture in the treatment of rheumatoid arthritis remains controversial.Therefore,the purpose of this study is to evaluate and explore the effect of Tiaodu Tongmai acupuncture in the treatment of preclinical rheumatoid arthritis of different ages through the generalized estimating equation,and to provide a basis for the application of acupuncture in rheumatoid arthritis. OBJECTIVE:To investigate the effect of Tiaodu Tongmai acupuncture therapy on preclinical rheumatoid arthritis(pre-RA)patients at different ages based on generalized estimating equation. METHODS:A total of 123 patients with preclinical rheumatoid arthritis treated from January to September 2023 were selected as the study objects and divided into study group(n=64)and control group(n=59)according to different treatment methods.The study group was given Tiaodu Tongmai acupuncture treatment,and the control group was given acetaminophen tablets.The baseline balance was adjusted by propensity score matching method.The clinical efficacy and cytokine levels before and after treatment between the two groups were compared.The generalized estimating equation model was established to evaluate the efficacy of Tiaodu Tongmai acupuncture therapy on preclinical rheumatoid arthritis patients at different ages. RESULTS AND CONCLUSION:(1)After 0 days of treatment,there were significant differences in joint pain and C-reactive protein expression between study and control groups(P＜0.05).After 4 weeks of treatment,there were significant differences in visual analogue scale scores,joint pain,C-reactive protein and erythrocyte sedimentation rate between the two groups(P＜0.05).After 8 weeks of treatment,there were significant differences in visual analogue scale scores,C-reactive protein and erythrocyte sedimentation rate between the two groups(P＜0.05).After 12 weeks of treatment,there were significant differences in visual analogue scale scores,C-reactive protein and erythrocyte sedimentation rate between the two groups(P＜0.05).(2)The total effective rate of the study group was 93.75%,while that of the control group was 79.17%.The clinical efficacy of the study group was significantly better than that of the control group(P＜0.05).(3)There were significant differences in interleukin-6,interferon-γ,macrophage migration inhibitory factor,rheumatoid factor IgA,rheumatoid factor IgM,metallomatrix proteinase 3,metallomatrix proteinase 9,and anti-cyclic citrulline antibody in the study group before and after treatment(P＜0.05).The levels of interleukin-6,interferon-γ,rheumatoid factor IgA,rheumatoid factor IgM,metallomatrix proteinase 3,metallomatrix proteinase 9,and anti-cyclic citrulline antibody in the control group after treatment were significantly different from those before treatment(P＜0.05).There were significant differences in interleukin-6,interferon-γ,macrophage migration inhibitory factor,rheumatoid factor IgA,rheumatoid factor IgM,metallomatrix proteinase 3,and anti-cyclic citrulline antibody between the two groups after treatment(P＜0.05).(4)After 12 weeks of treatment,the comprehensive efficacy of patients of all ages in the study group was better than that of the control group(P＜0.05).After 8 weeks of treatment,the comprehensive efficacy of patients aged 23-35,36-50,and 51-60 years old in the study group was better than that of the control group(P＜0.05),and the comprehensive efficacy of patients aged 18-22 years old was comparable between the two groups.After 4 weeks of treatment,the comprehensive efficacy of patients aged 36-50 and 51-60 years old in the study group was better than that of the control group(P＜0.05),and the comprehensive efficacy was comparable between the two groups of patients aged 18-22 and 23-35 years.Overall,Tiaodu Tongmai acupuncture therapy has advantages in treating preclinical rheumatoid arthritis patients aged 36-50 and 51-60 years.

This article reviews relevant literature on the prevention and treatment of cancer with hesperidin published in the past 10 years by searching electronic databases such as China National Knowledge Infrastructure（CNKI）， Wanfang， and PubMed， and summarizes the research progress on the anticancer mechanism of hesperidin. Hesperidin has a wide range of pharmacological effects， including anti-inflammatory， antioxidant， antibacterial， antiviral， anticancer， immune-regulatory， anti-radiation， neuroprotective and cardiovascular protective properties and so on. Its anticancer mechanisms mainly include inhibiting cancer cell proliferation， promoting apoptosis， reducing angiogenesis， inhibiting invasion and migration of cancer cells， regulating immunity and autophagy， and exerting antioxidant and anti-inflammatory effects. As a broad-spectrum anticancer drug， hesperidin manifests chemo-preventive and therapeutic effects across various cancers， contingent upon its multifaceted anticancer mechanisms. Furthermore， this article summarizes the synergistic effects of hesperidin in combination with cisplatin， doxorubicin， cyclophosphamide and paclitaxel. It elucidates that hesperidin can enhance the cytotoxicity of these anticancer drugs against cancer cells while mitigating drug resistance and adverse side effects. Nonetheless， the clinical use is somewhat constrained due to its poor water solubility and limited bioavailability. Therefore， this article also outlines the current strategies for enhancing hesperidin's bioavailability， including structural modification， combination with other chemical substances， and utilization of nano drug carriers.The discovery of derivatives of hesperidin not only preserves the anticancer efficacy of hesperidin, but also effectively overcomes the shortcomings of poor water solubility and low bioavailability of hesperidin, effectively predicting the good application prospects of hesperidin and its derivatives.

Objective To study the mechanism of Jiawei Chengqi decoction combined with enteral nutrition emulsion in the treatment of stroke patients by regulating brain-derived neurotrophic factor/tropomyosin kinase receptor B(BDNF/TrkB)signaling pathway.Methods A total of 200 stroke patients treated in the intensive care unit(ICU)of Xingtai Third Hospital from January 2020 to October 2022 were selected as the study objects,and the patients were randomly divided into observation group and control group,with 100 cases in each group.The control group was treated with enteral nutrition emulsion,and the observation group was treated with Jiawei Chengqi decoction on this basis[drug composition:Raw Rhubarb 10 g(bottom),Raw Malt 30 g,Sodium Powder 6 g(flush),Substitute Ochre 30 g(first),Magnolia Officinalis 10 g,Stir-fried Raphanus Japonicus 15 g,Stir-fried Fructus Aurifolius 15 g,Spiral Flower 10 g(pan-fried),whole Trichosanthes 30 g.100-200 mL of juice in water was consumed in 2 separate doses,once daily].The clinical efficacy,nutritional indexes[total protein(TP),hemoglobin(Hb),prealbumin(PA),albumin(ALB)],intestinal mucosal barrier indexes[diamine oxidase(DAO),procalcitonin(PCT),D-lactic acid],traditional Chinese medicine(TCM)syndrome score,TrkB,BDNF,and calcitonin gene-related peptide(CGRP)level differences were compared between the two groups after treatment.Results The total clinical effective rate of the observation group was significantly higher than that of the control group[92.00%(92/100)vs.81.00%(81/100),P<0.05].The levels of nutritional indexes TP,Hb,ALB,intestinal mucosal barrier,TCM syndrome score,TrkB,BDNF and CGRP in the two groups after treatment were significantly lower than those before treatment,and the nutritional indexes TP,Hb and PA in the observation group were significantly higher than those in the control group after treatment[TP(g/L):68.09±10.28 vs.64.53±9.81,Hb(g/L):133.48±33.38 vs.121.38±30.72,PA(mg/L):231.29±22.75 vs.219.73±20.48,all P<0.05],the levels of nutrition index ALB,mucosal barrier index,TCM syndrome score,TrkB,BDNF and CGRP were significantly lower than those of control group[ALB(g/L):36.78±5.32 vs.39.49±5.48,DAO(kU/L):3.19±0.53 vs.3.91±0.59,PCT(μg/L):5.05±0.68 vs.6.14±0.89,D-lactic acid(mmol/L):0.14±0.03 vs.0.22±0.03,abdominal pain and diarrhea scores:1.34±0.62 vs.2.45±0.73,abdominal cold pain scores:1.63±0.63 vs.2.38±1.03,waist and knee tenderness scores 0.96±0.41 vs.1.75±0.52,diet scores:1.14±0.41 vs.1.58±0.69,cold body cold integral scores:0.97±0.42 vs.1.68±0.54,TrkB(ng/L):285.02±28.15 vs.375.84±36.12,BDNF(ng/L):1 306.84±244.83 vs.1 718.84±378.83,CGRP(ng/L):79.68±9.56 vs.87.27±11.01,all P<0.05].Conclusion Using Jiawei Chengqi decoction in conjunction with enteral nutrition emulsion for treating stroke patients in the ICU can significantly improve the intestinal mucosal indexes and nutritional indexes of patients,making it a promoting approach in clinical practice.

Objective To investigate the relationship between the vaginal microecology and CC chemokine ligand 28(CCL28)expression levels in the vaginal lavage fluid of patients with persistent high-risk human papillomavirus(HR-HPV)infection.Methods A total of 132 patients with HR-HPV infection at Tangshan Maternal and Child Health Hospital from April 2021 to April 2023 were selected as study subjects.Patients with HR-HPV infection were categorized into the single infection group and the persistent infection group according to the review results.The CCL28 and vaginal microecological indicators of the two groups were compared.Logistic regression was used to analyze the risk factors for persistent HR-HPV infection.The dose-response relationship between CCL28 and persistent HR-HPV infection was analyzed by fitting restricted cubic splines with logistic regression.Spearman and multiple linear regression were used to analyze the relationship between CCL28 and vaginal microecological indicators.Results After the re-examination of patients,there were 84(63.64％)cases of single infection and 48(36.36％)cases of persistent infection.The percentage of colony density grade Ⅰ(54.17％vs 16.67％),colony diversity grade Ⅰ(47.92％),pH＞4.5(47.92％),hydrogen peroxide positivity(54.17％),and vaginal microecological imbalance(66.67％),and the level of CCL28[83.63(77.11,99.62)ng/L]in the persistent infection group were higher than those of the single infection group[16.76％,21.43％,15.48％,30.95％,41.67％,71.11(58.12,82.16)ng/L],and the differences were statistically significant(x2/Z=5.064～20.338,all P＜0.05).Logistic regression analysis showed that colony density grade Ⅰ(OR=3.556,95％CI:1.241～10.194,P=0.018),PH＞4.5(OR=4.366,95％CI:1.164～16.380,P=0.029),and high CCL28 levels(OR=1.048,95％CI:1.018～1.080,P=0.002)were independent risk factors for persistent infection in HR-HPV-infected patients.Logistic regression fitting restricted cubic splines showed a linear relationship between CCL28 and persistent HR-HPV infection(nonlinear test x2=0.61,P=0.437).Spearman correlation analysis results showed that CCL28 was positively correlated with age(r=0.205,P=0.018),colony density(r=0.429,P＜0.001),colony diversity(r=0.389,P＜0.001),pH(r=0.357,P＜0.00l),hydrogen peroxide(r=0.409,P＜0.001),leukocyte esterase(r=0.392,P＜0.00l),and vaginal microecology(r=0.473,P＜0.001).Linear regression analysis showed that colony density(β=12.295,P=0.003)and colony diversity(β=9.196,P=0.032)were independently associated with CCL28.Conclusion Colony density and colony diversity were independently correlated with CCL28.Colony density grade Ⅰ,pH＞4.5,and high CCL28 levels suggest an increased risk of persistent infection in patients with HR-HPV infection.

OBJECTIVE: To observe the clinical effect and safety of fire needle therapy combined with bladder function training on neurogenic bladder (NB) caused by spinal cord injury. METHODS: A total of 60 patients with NB caused by spinal cord injury were randomly divided into an observation group and a control group , with 30 cases in each group. On the basis of conventional treatment with western medicine, the bladder function training was adopted in the control group, once a day and for 4 weeks. In the observation group, on the basis of the interventions as the control group, fire needling was operated at bilateral Sanyinjiao (SP 6) and Guanyuan (CV 4) and Zhongji (CV 3), once every two days and for 4 weeks (14 interventions in total). Separately, at the baseline and in 2 and 4 weeks of interventions, the urination conditions (average daily urination frequency, average daily leakage frequency, average daily single urination volume) were recorded in the two groups; the urodynamic parameters (maximum flow rate [Qmax], maximum detrusor pressure at maximum flow rate [PdetQmax], residual urine volume [RUA], maximum cystometric capacity [MCC], and bladder pressure) were detected; the neurogenic bladder symptom score (NBSS), urinary symptom distress score (USDS) were observed. Before and after treatment, the score of World Health Organization quality of life assessment scale-brief (WHOQOL-BREF) was observed in the two groups. The therapeutic effect, the incidence of urinary infection, and the safety were evaluated. RESULTS: In 2 and 4 weeks of interventions, the average daily urination frequency, the average daily leakage frequency, RUA, and the scores of NBSS and USDS decreased in the two groups when compared with the baseline (P<0.05). In 4 weeks of interventions, the above-mention outcomes were lower than those in 2 weeks of interventions (P<0.05); and the results in the observation group were lower in 2 and 4 weeks of interventions when compared with the control group (P<0.05). In 2 and 4 weeks of interventions, the average daily single urination volume, Qmax, PdetQmax, MCC, and bladder pressure increased in the two groups compared with the baseline (P<0.05). In 4 weeks of interventions, the above-mention outcomes were elevated in comparison with those in 2 weeks of interventions (P<0.05); and except for bladder pressure, the results in the observation group in 2 and 4 weeks of interventions were higher when compared with the control group (P<0.05). In 4 weeks of interventions, the scores of each dimension and the total scores of WHOQOL-BREF increased in comparison with the baseline in the two groups (P<0.05), and the scores of the observation group were higher than those of the control group (P<0.05). The incidence of urinary infection was 0% (0/30) in the observation group and 10.0% (3/30) in the control group, without significant difference (P>0.05). The total effective rate of the observation group was 93.3% (28/30), which was higher than that (73.3%, 22/30) of the control group (P<0.05). No serious adverse reactions occurred in the patients of the observation group. CONCLUSION Fire needle therapy combined with bladder function training can effectively relieve the clinical symptoms, ameliorate urination, restore bladder function and improve the quality of life in the patients with NB caused by spinal cord injury. This therapeutic regimen presents a high safety in practice.
Humans ; Spinal Cord Injuries/therapy* ; Female ; Male ; Acupuncture Therapy ; Middle Aged ; Adult ; Urinary Bladder, Neurogenic/etiology* ; Urinary Bladder/physiopathology* ; Urination ; Young Adult ; Treatment Outcome ; Aged ; Adolescent ; Combined Modality Therapy