1.Effects of Zhimu (Anemarrhena asphodeloides)-Huangbai (Phellodendron amurense) Medicinal Pair on Femoral Microstructure and Osteogenic-Adipogenic Differentiation in Ovariectomized Osteoporosis Model Rats
Chuncai LI ; Mingxing YUAN ; Jiawei LI ; Jing DENG ; Chongyang SHEN ; Yuan LIU
Journal of Traditional Chinese Medicine 2025;66(16):1704-1710
ObjectiveTo investigate the potential mechanisms of Zhimu (Anemarrhena asphodeloides)-Huangbai (Phellodendron amurense) medicinal pair in alleviating postmenopausal osteoporosis (PMOP). MethodsSixty unpregnant female SD rats were randomly divided into five groups, blank group, model group, low-dose Zhimu-Huangbai group, high-dose Zhimu-Huangbai group, and estradiol group, with 12 rats in each group. Except for the blank group, all other groups had their ovaries removed to create PMOP rat models, while the blank group only had the fat tissue around the ovaries removed. One week after the ovarian removal, the low-dose and high-dose Zhimu-Huangbai groups received concentrated solution of Zhimu and Huangbai with 1.8, 7.2 g/(kg·d) via gavage, the estradiol group received estradiol solution 0.09 mg/(kg·d) via gavage, and the blank group and the model group received 10 ml/(kg·d) of normal saline via gavage, once daily for 12 weeks. Before sampling, the body mass of the rats was recorded, and uterine tissue was taken to calculate the uterine index. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum were detected by ELISA; micro CT was used to examine the parameters of femoral microstructure, including bone volume/tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), bone mineral density (BMD), trabecular separation (Tb.Sp), and cortical bone area (Ct.Ar). HE staining was used to observe pathological changes in the femur; RT-qPCR was used to detect the mRNA expression of osteogenic-adipogenic differentiation-related factors in femoral tissue, including Runt-related transcription factor 2 (Runx2), bone morphogenetic protein 2 (BMP-2), peroxisome proliferator-activated receptor γ (PPARγ), chemerin and chemokine-like receptor 1 (CMKLR1). ResultsCompared with the blank group, the model group showed a significant increase in body mass, a significant decrease in the uterine index, BV/TV, Tb.N, Tb.Th and BMD, a significant increase in Tb.Sp, and serum IL-1β, IL-6, and TNF-α levels, a significant reduction of mRNA expression of Runx2 and BMP-2 in bone tissue, and a significant increased mRNA expression of PPARγ, chemerin, and CMKLR1 (P<0.01). HE staining revealed that the femoral tissue showed a reduction and sparsity of trabeculae, a significant enlargement of the medullary cavity, and a large number of fat cells. Compared to the model group, the low-dose, high-dose Zhimu-Huangbai groups, and estradiol group showed significant improvements in all the above-mentioned indicators (P<0.05 or P<0.01). HE staining revealed a significant increase in trabeculae, more organized arrangement, and a marked reduction in fat cells. Compared to low-dose Zhimu-Huangbai group, the high-dose Zhimu-Huangbai group exhibited a significant increase in the uterine index and BMD, and a significant reduction in body mass and PPARγ and Chemerin mRNA expression (P<0.05 or P<0.01). Compared to high-dose Zhimu-Huangbai group, the estradiol group showed a decrease in uterine index, BV/TV, Tb.N, Tb.Th, BMD, and BMP-2 mRNA expression, while the levels of IL-1β, TNF-α, and IL-6, as well as Tb.Sp and the mRNA expressions of PPARγ, chemerin, and CMKLR1 increased (P<0.05 or P<0.01). ConclusionThe Zhimu-Huangbai medicinal pair can alleviate PMOP bone loss, and its mechanism of action is related to reducing the levels of inflammatory factors, correcting the disorder of osteogenic-adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and promoting the differentiation of BMSCs into osteoblasts.
2.Discussion on Differentiation and Treatment of Sudden Hearing Loss by Traditional Chinese Medicine Based on the Classification of Hearing Curve
Xinru WANG ; Yang LI ; Jiajia ZHANG ; Yan MENG ; Chongyang ZHANG ; Xueshi DI ; Zhiwei FENG ; Junjie LIANG ; Peng BAI
Journal of Traditional Chinese Medicine 2024;65(11):1126-1131
Pure-tone audiometry can be performed to evaluate the type and degree of hearing loss, whose results can be divided into four types including low-frequency descending, high-frequency descending, flat descending and total deafness. The low-frequency descending type of sudden hearing loss (SHL) is more likely to be spleen deficiency and dampness exuberance, the high-frequency descending type is often due to yin deficiency of liver and kidney, the flat descending type is commonly associated with qi and blood depletion, and the type of total deafness is often linked to blood stasis. Our team has further developed a comprehensive diagnostic and therapeutic approach for SHL, emphasizing "the integration of disease and syndrome, the combination of acupuncture and herbal medicine, and dynamically administering treatment". Firstly, it advocates integrating disease diagnosis with syndrome differentiation. Secondly, it recommends combining acupuncture and herbal medicine, with local acupoints such as Ermen (TE 21), Tinggong (SI 19), Tinghui (GB 2), and Yifeng (TE 17) used to unblock the auditory orifice, and herbal prescriptions tailored to the hearing curve patterns. For the low-frequency descending type, it is recommended to fortify the spleen and percolate dampness by taking distal points of spleen channel and stomach channel and using Shenling Baizhu Powder (参苓白术散). For the high-frequency descending type, the method of nourishing kidney and calming liver is recommended, using distal points of kidney and liver channels and taking Erlong Zuoci Pills (耳聋左慈丸). Regarding the flat descending type, tonifying qi and nourishing blood is advised, for which acupoints of Conception Vessel, spleen, stomach and large intestine channels can be needled, and Yiqi Congming Decoction (益气聪明汤) can be administered. For the total deafness type, it is recommended to activate blood and dissipate stasis, often with Xuehai (SP 10) and Geshu (BL 17) needled and Tongqiao Huoxue Decoction (通窍活血汤) administered. All these are conducted to treat the root and branch simultaneously. At the same time, it emphasizes the need to consider complex syndrome presentations and disease progression, dynamically analyze the disease causes and mechanisms, and adjust treatment according to the changing syndromes. In conclusion, this article is expected to inspire clinical diagnosis and treatment of SHL.
3.Observation of acute clinical manifestations of cosmetology-related ocular damage
Jian JIAO ; Wen HUA ; Chongyang SHE ; Weiwei ZHU ; Xuedong LI
International Eye Science 2024;24(7):1152-1156
AIM: To investigate the acute clinical manifestations of cosmetology-related ocular damage(COD).METHODS:Retrospective study. A total of 53 cases(89 eyes)with ocular damage caused by cosmetology from April 2016 to October 2021 were collected. The clinical features were analyzed, including age, gender, affected eye(s), clinical manifestations, injury cause, treatment procedures, and prognosis.RESULTS: All 53 patients were female, aged 22-45 years, with an average age of 28.4±6.7 years. Monocular injuries were observed in 17 patients, and binocular injuries in 36 patients. The same eye could exhibit two or more ocular damage simultaneously. The primary cosmetology procedures causing COD were eyeliner tattooing(38 eyes; 43%), eyelash extensions(18 eyes; 20%), removal of false eyelashes(11 eyes; 12%), mascara application(8 eyes; 9%), double eyelid surgery(6 eyes; 7%), and others(8 eyes; 9%). Major ocular damages included corneal damage(56 eyes; 63%), eyelid contact dermatitis(26 eyes; 29%), conjunctivitis(19 eyes; 21%), reactive eyelid edema(13 eyes; 15%), ocular surface foreign bodies(12 eyes; 14%), bacterial infection of the palpebral margin(10 eyes; 11%), and others(5 eyes; 6%). These 5 eyes included 1 eye(1%)with central retinal artery occlusion caused by periocular injection of hyaluronic acid. The majority of patients(74 eyes)recovered within 1-2 wk with appropriate treatment, while filamentosa keratitis appeared in 3 eyes and the eye with central retinal artery occlusion had poor prognosis.CONCLUSIONS: COD predominantly occurs in young and middle-aged females with cosmetology experience. The most common cosmetology procedure leading to COD is eyeliner tattooing, and corneal damage is the most significant type of COD. COD can be effectively prevented and treated, resulting in a generally favorable prognosis.
4.Neck Seven-Line Method Combined with Periauricular Acupuncture for 33 Patients with Sudden Hearing Loss in Non-Acute Stage: A Randomized Controlled Trial
Chongyang ZHANG ; Junjie LIANG ; Yang LI ; Xinru WANG ; Yu XING ; Xueshi DI ; Wenting SUN ; Peng BAI
Journal of Traditional Chinese Medicine 2024;65(15):1571-1577
ObjectiveTo evaluate the effectiveness and safety of neck seven-line method combined with periauricular acupuncture as salvage treatment for sudden hearing loss in non-acute stage. MethodsSixty-six patients with non-acute stage of sudden hearing loss with a disease duration of 15-90 days were randomly divided into 33 cases each in treatment group and control group. The treatment group was given neck seven-line method combined with acupuncture at periauricular points; the control group used sham acupuncture and sham electroacupuncture at the same points. Both groups were treated 3 times a week for 6 weeks. The pure tone average hearing threshold of impaired frequencies were examined before treatment, after treatment finish and at follow-up (week 10), and the difference between pure tone average hearing threshold of impaired frequencies before and after treatment was calculated; calculate the proportion of patients with ≥10 dB improvement in pure tone average hearing threshold of impaired frequency after treatment and at follow-up; compare the patients' Tinnitus Evaluation Scale (TEQ) scores, Chinese Medicine Quality of Life Assessment Scale (CQ-11D) health utility values and the difference between before and after treatment, and record the occurrence of adverse events. ResultsThe pure tone average of impaired frequency in the treatment group were (50.57±18.07) dB and (47.70±17.42) dB at post-treatment and follow-up respectively, and (54.38±21.77) dB and (53.36±20.99) dB in the control group at post-treatment and follow-up. Compared with the pre-treatment period, the pure tone average hearing threshold of impaired frequency in the two groups significantly decreased (P<0.05) at post-treatment and follow-up. The difference of pure tone average hearing threshold of impaired frequency in the treatment group after treatment and at the follow-up visit compared to that before treatment was lower than those in the control group (P<0.05). After treatment, 13 patients (39.39%) in the treatment group and 10 patients (30.30%) in the control group showed improvement of ≥10 dB in pure tone average hearing threshold of impaired frequency; at the follow-up visit, 18 patients (54.55%) in the treatment group and 10 patients (30.30%) in the control group showed improvement of ≥10 dB in the pure tone average hearing threshold of impaired frequency, and there was no statistical significance for comparison between groups at the time of post-treatment and follow-up (P>0.05). The TEQ score of the treatment group significantly lower than that before treatment (P<0.05); the TEQ score of the treatment group and the difference between before and after treatment significantly lower than that of the control group (P<0.05). The difference in CQ-11D health utility values and the difference between before and after treatment were not statistically significant between the two groups (P>0.05). Eight cases of acupuncture-related adverse events occurred among 66 patients, including subcutaneous haematomas after needling, severe pain during needling and needle fainting, which disappeared after symptomatic treatment and did not affect the following treatment. ConclusionNeck seven-line method combined with periauricular acupuncture could be used as salvage treatment for sudden hearing loss in non-acute stage to improve the pure tone average hearing threshold of impaired frequency, alleviate tinnitus, with safety.
5.Prenatal diagnosis analysis of three cases of Turner syndrome fetuses with complex mosaic small supernumerary marker chromosomes
Chongyang ZHU ; Chunxu LI ; Peipei XU ; Ling LIU
Chinese Journal of Medical Genetics 2024;41(11):1363-1370
Objective:To explore the value of applying multiple genetic testing techniques for the prenatal diagnosis of Turner syndrome fetuses with complex mosaic small supernumerary marker chromosomes (sSMC).Methods:Chromosomal karyotypes of amniotic fluid samples from 5 030 pregnant women who had undergone amniocentesis at the Prenatal Diagnosis Center of the Third Affiliated Hospital of Zhengzhou University from January to December 2022 were retrospectively reviewed. Three fetuses with complex mosaicism fetuses (carrying 2 types of sSMC) were selected as the study subjects. Genetic tests including G-banded chromosomal karyotyping analysis, fluorescence in situ hybridization (FISH), chromosomal microarray analysis (CMA), and copy number variation sequencing (CNV-seq) were used to clarify the origin and mosaic status of the sSMC. This study has been approved by the Medical Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (No. 2023-159-01).Results:G-banded chromosomal analysis of fetus 1 showed a karyotype of 45, X[64]/46, X, + mar1[13]/46, X, + mar2[3]. FISH results showed that 52% of of its cells had contained one X chromosome signal, whilst 48% contained two X chromosome signals. CMA results revealed the fetus had harbored a 32.32 Mb and a 50.93 Mb deletion in Xp22.33p21.1 and Xq22.2q28 regions, respectively, in addition with mosaic deletions of approximately 1.43 copies, 1.78 copies and 1.43 copies in the Xp21.1p11.1, Xq11.1q21.1 and Xq21.2q22.2 regions, respectively. The fetus 2 had a karyotype of 45, X[27]/46, X, + mar1[14]/46, X, + mar2[12]. FISH results indicated that 88% of its cells contained one X chromosomes signal and two Y chromosome signals, and 12% contained signals for one X chromosomes signal and one Y chromosome signal. CNV-seq results revealed a deletion of 7.74 Mb in the Yq11.222q11.23 region and a mosaic duplication of approximately 1.738 copies in the Yp11.31q11.221 region. The fetus 3 had a karyotype of 45, X[60]/46, X, + mar1[11]/46, X, + mar2[6]. FISH results showed that 28% of its cells contained one X chromosome signal, and 72% contained tow X chromosome signals. CNV-seq results revealed deletions of 55.60 Mb and 53.50 Mb in the Xp22.33p11.1 and Xq22.1q28 regions, respectively, along with a mosaic deletion of approximately 1.85 copies in the Xp11.1q13.2 region and a mosaic repeats of approximately 2.66 copies in the Xq13.2q22.1 region. The sSMCs in the 3 fetuses had all originated from sex chromosomes and were of complex mosaic type. After genetic counseling, the three couples had all opted to terminate the pregnancy.Conclusion:The combined use of multiple genetic testing techniques has determined the origin and structure of complex mosaic sSMCs and provided a basis for prenantal diagnosis and genetic counseling.
6.Synthesis of a 68Ga-labeled bicyclic peptide targeting Nectin-4 and its application research in breast cancer imaging
Liqi LI ; Yue XU ; Donghui PAN ; Junjie YAN ; Xinyu WANG ; Chongyang CHEN ; Lizhen WANG ; Min YANG ; Yuping XU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(12):741-747
Objective:To prepare a novel 68Ga-labeled bicyclic peptide targeting poliovirus receptor related protein 4 (PVRL4, Nectin-4), and evaluate its feasibility for breast cancer imaging via in vitro and in vivo experiments. Methods:A Biotin-modified bicyclic peptide targeting Nectin-4, Biotin-BMIC, was synthesized, and its targeting properties were preliminarily evaluated by in vitro cell staining experiments. BMIC was modified by 1, 4, 7-triazonane-1, 4-diacetic acid (NODA) and the labeling precursor NODA-BMIC was prepared. A potential PET probe targeting Nectin-4, 68Ga-NODA-BMIC was prepared by one-step labeling strategy. The imaging properties of the probe were investigated by in vivo microPET imaging and in vitro experiments in mice bearing breast tumors. Data were analyzed by independent-sample t test and repeated measures analysis of variance. Results:Fluorescence staining of the cells showed that the fluorescently labeled bicyclic peptide, Biotin-BMIC, was highly aggregated in Nectin-4 positive BT474 breast cancer cells compared to those in Nectin-4 negative MDA-MB-231 cells. The uncorrected yield of 68Ga-NODA-BMIC was (71.5±2.2)% and the radiochemical purity was greater than 95%. The specific activity was greater than 3 GBq/μmol. After incubation 10, 30, 60 and 120 min, higher radioactivity uptakes were found in BT474 breast cancer cells compared to those in MDA-MB-231 breast cancer cells respectively ( F=1 302.00, P<0.001). MicroPET imaging showed that the BT474 xenograft tumors were clearly visible with favorable contrast. A significant statistical difference in uptakes between BT474 and MDA-MB-231 xenograft tumor uptake at 10, 30, 60, and 120 min after probe injection respectively was existed ( F=1 826.00, P<0.001). At 60 min postinjection, the uptake value of BT474 tumors was (5.03±0.14) percentage activity of injection dose per gram of tissue (%ID/g), which was significantly higher than that of MDA-MB-231 tumors ((0.19±0.04) %ID/g; t=79.40, P<0.001). Meanwhile, the tumor-to-muscle ratios in the former were also greater than those in the latter ( F=222.00, P<0.001). At 60 min postinjection, the tumor-to-muscle ratio in the former was significantly higher than that in the latter (24.75±3.10 vs 1.30±0.15; t=14.31, P=0.002). The results were consistent with the immunohistochemistry staining. Conclusions:A novel bicyclic peptide PET probe targeting Nectin-4, 68Ga-NODA-BMIC, is easy to be synthesized and owns satisfactory labeling yield and radiological purity. The imaging performance is good and the target tissues could be visualized. It may play a unique role in the diagnosis and treatment of breast cancer.
7.The relationship between non-alcoholic fatty liver disease and hepatic fibrosis with skeletal muscle mass in patients with type 2 diabetes mellitus
Xinyuan GUO ; Mei HAN ; Dengrong MA ; Xiaohui ZAN ; Yangting ZHAO ; Xiaoyu LYU ; Kai LI ; Chongyang CHEN ; Yawen WANG ; Jingfang LIU
Chinese Journal of Endocrinology and Metabolism 2024;40(8):660-668
Objective:To investigate the relationship between non-alcoholic fatty liver disease(NAFLD) and hepatic fibrosis and skeletal muscle mass in patients with type 2 diabetes mellitus(T2DM).Methods:A total of 685 T2DM patients diagnosed at the Endocrinology department of Lanzhou University First Hospital, from April 2022 to May 2023, were divided into NAFLD and Non-NAFLD groups, and the NAFLD group was further categorized into fibrosis and non-fibrosis based on aspartate aminotransferase(AST) /alanine aminotransferase(ALT) level. The differences in appendicular skeletal muscle mass(ASM), appendicular skeletal muscle mass index(ASMI), and the prevalence of muscle mass loss were compared across groups. The correlations between ASMI and NAFLD, as well as liver fibrosis were analyzed by binary logistic regression.Results:Among male T2DM patients, those with NAFLD had lower ASMI levels and a higher prevalence of muscle mass reduction compared to non-NAFLD group. Among female T2DM patients, those with NAFLD had lower levels of ASM and ASMI, and a higher prevalence of muscle mass reduction compared to non-NAFLD group. ASMI levels in both male and female T2DM patients were independently negatively correlated with NAFLD risk( OR=-0.696, 95% CI 0.579-0.837; OR=-0.757, 95% CI 0.629-0.911). In NAFLD patients, ASM and ASMI levels were lower in those with liver fibrosis compared to those without fibrosis; however, the prevalence of muscle mass reduction did not differ significantly. Among male NAFLD patients, ASMI levels were independently negatively correlated with the risk of liver fibrosis( OR=-0.726, 95% CI 0.537-0.983), while no correlation was found in female patients. Conclusion:Reduced muscle mass is independently associated with the risk of NAFLD in both male and female T2DM patients. In males, reduced muscle mass is also independently related to the risk of liver fibrosis.
8.Effects of non-pharmaceutical interventions on epidemiological characteristics of respiratory pathogens in adults
Xue YANG ; Chongyang WU ; Li XIONG ; Mengjiao LI ; Yu YUAN ; Yuling LIN ; Yuling XIAO ; Yi XIE
International Journal of Laboratory Medicine 2024;45(12):1425-1430
Objective To explore the impact of non-pharmaceutical interventions(NPIs)on the prevalence of respiratory pathogens in adults,and to understand the scientific value and long-term effect of NPIs.Methods A retrospective study was conducted to collect the clinical data and laboratory examination data of adult patients with respiratory tract infection in West China Hospital,Sichuan University from 2017 to 2023,and the patho-gen,population,season and other aspects were analyzed in different periods.The analysis period included 2017 to 2019(before the implementation of NPIs),2020 to 2022(during the implementation of NPIs),and January to December 2023(after the implementation of NPIs).Results A total of 33 068 adult patients with respira-tory tract infection were included.The overall prevalence of 8 adult respiratory pathogens from 2017 to 2019(26.95%)was higher than that from 2020 to 2022(8.70%),and the difference was statistically significant(P<0.05).There were significant differences in the prevalence of pathogens among different genders,ages and seasons in the first,middle and last three periods of NPIs implementation(P<0.05).Before the imple-mentation of NPIs,the seasonal peak of respiratory prevalence appeared from January to March each year.With the implementation of NPIs,the seasonal peak of respiratory prevalence appeared from January to March 2020(10.09%),October to December 2021(9.32%),July to September 2022(15.23%),respectively.After the implementation of NPIs,the seasonal peak of respiratory prevalence appeared from October to December 2023(21.20%).Among the 8 pathogens,the change of prevalence of influenza A virus H1N1(2009)was the most obvious,and the prevalence was 17.42%,0.00%and 6.99%before,during and after the implementation of NPIs,respectively.Conclusion Due to the influence of NPIs and other factors,the epidemic characteristics of respiratory pathogens have changed from 2017 to 2023.Attention to the emerging characteristics of patho-gen prevalence is important for the prevention,diagnosis and control of respiratory infectious diseases during public health emergencies.
9.Use situation of Rush among men who have sex with men in Chongqing City and its influencing factors analysis
Hui LONG ; Chongyang BAI ; Long LI ; Guohui WU ; Tianyu TAN
Chongqing Medicine 2024;53(20):3145-3149
Objective To understand the use situation of Rush among the men who have sex with men(MSM)in Chongqing City and its influencing factors,so as to provide a basis for the formulation of relevant prevention and control measures.Methods The cross-sectional survey was adopted.The MSM population in Chongqing City were recruited through the network recruitment and referral of social organizations to conduct an anonymous electronic questionnaire survey to obtain the information such as demographic characteristics,sexual partners and sexual behavior characteristics in the past 6 months,Rush use,HIV and syphilis antibody testing.The X2 test and non-conditioned multi-factor Logistic regression analysis were used to analyze the in-fluencing factors of Rush use.Results A total of 494 MSM were surveyed,the Rush usage rate was 22.1%.The results of the multi-factor logistic regression analysis showed that the age of first time homosexuality be-havior<18 years old(OR=2.715,95%CI:1.353-5.451),unprotected homosexual anal intercourse in near-ly 6 months(OR=2.237,95%CI:1.355-3.693),having multiple same-sex partners in nearly 6 months(OR=3.610,95%CI:1.449-8.993),same-sex casual partners in the nearly 6 months(OR=3.734,95%CI:2.221-6.280)were the risk factors for MSM using Rush.Conclusion The RUSH use rate among the MSM population in Chongqing City is high.The targeted intervention measures should be carried out,especially to strengthen the propaganda and education of the population who first have homosexual behavior under 18 years old,have unprotected homosexual anal intercourse in the nearly 6 months,have multiple homosexual perma-nent partners in the last 6 months,and have homosexual temporary partners,in order to reduce the use rate of Rush and thereby reduce the risk of HIV infection and transmission in this population.
10.Chronic myeloid leukemia with e6a2 fusion gene: a case report and literature review
Jingjing LI ; Lingling YUE ; Pengyun ZENG ; Chongyang WU ; Huiling CHEN
Chinese Journal of Hematology 2024;45(3):299-302
Chronic myeloid leukemia (CML) with e6a2 transcript type is very rare in clinic,which is usually related to disease aggressiveness. Its clinical characteristics and relationship with tyrosine kinase inhibitor efficacy are still unclear. In this paper, the clinical characteristics and related laboratory tests of a patient with e6a2 fusion gene positive CML characterized by multiple osteolytic bone destruction throughout the body and eosinophil infiltration in gastrointestinal tract, lymph nodes and other organs were retrospectively analyzed, and the relevant literature was reviewed. The patient was Ph chromosome positive with chromosome +8, and the common BCR::ABL1 transcript of CML was negative, but e6a2 transcript was positive detected by RT-PCR. The patient was treated with dasatinib 100 mg/d. Three months later, the patients achieved CHR, CCyR and MR4.0. However, the e6a2 transcript is very rare in clinical practice, and more cases of e6a2 transcript need to be studied to clarify its clinical characteristics and improve the treatment effect of these rare cases.

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