1.Biocontrol Effect and Culture Condition Optimization of Trichoderma citrinoviride Strain Isolated from Rhizosphere of Atractylodes lancea
Kuo LI ; Sheng WANG ; Hongyang WANG ; Zheng PENG ; Xiuzhi GUO ; Tielin WANG ; Yuefeng WANG ; Lanping GUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):192-199
ObjectiveRoot rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea. Trichoderma spp. are effective in the biocontrol of root rot without causing environmental pollution. This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A. lancea and to solve the problem of disease prevention and control in the planting and production of A. lancea. MethodTrichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing. The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments. The biocontrol potential of T2204 on root rot of A. lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A. lancea were investigated by pot experiments and GC-MS, respectively. In addition, the optimal medium, photoperiod, temperature, pH, and carbon and nitrogen sources for the culture of T2204 were explored. ResultThe Trichoderma isolate T2204 was identified as T. citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot. In the dual-culture experiments with the two pathogenic strains, T2204 showcased the inhibition rates of 77.90% and 76.80%, respectively. In the co-culture experiments with the two pathogenic strains, the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11% and 81.11%, respectively. The pot experiments showed that the survival rate of A. lancea seedlings infected by root rot reached 100% after inoculation with T2204 and was only 50% in the case without inoculation of T2204. After 150 days of cultivation, the dry weight and atractylodin content of the rhizome of A. lancea plants treated with T2204 increased by 32% (P<0.05) and 11%, respectively, compared with the untreated group. The optimal conditions for the growth of T2204 were PDA or PSA medium, photoperiod of 12 h dark/12 h light, 25-30 °C, pH 5-6, carbon sources of glucose, D-fructose, soluble starch, and maltose, and the nitrogen sources of ammonium sulfate and ammonium dihydrogen phosphate. The optimal conditions for the sporulation of T2204 were PSA or CMA medium, photoperiod of 12 h dark/12 h light, 20-30 °C, pH 8, carbon source of sucrose, and nitrogen source of sodium nitrate. ConclusionT2204 could improve the growth and root rot resistance of A. lancea and promote the accumulation of medicinal compounds. The findings laid a foundation for the industrialized production and application of T2204 in the production of A. lancea in the future.
2.Histopathological diagnosis of pulmonary sclerosing pneumocytoma in needle biopsy specimens
Qianqian SI ; Jing HAN ; Xianzheng GAO ; Yuanyuan ZANG ; Tielin WANG ; Shenglei LI
Chinese Journal of Pathology 2024;53(11):1105-1110
Objective:To investigate the diagnostic features of pulmonary sclerosing pneumocytoma (PSP) in needle biopsy specimens so as to improve the preoperative diagnostic accuracy and to prevent misdiagnoses.Methods:A total of 79 needle biopsy cases confirmed as PSP in surgical resection specimens were collected in the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China from January 2015 to January 2023. A retrospective analysis was conducted to investigate the clinical, pathological, and immunohistochemical characteristics of PSP.Results:Among the 79 cases, there were 8 males and 71 females, with an age range of 14 to 67 years (median 47 years). Among the 79 needle biopsy cases of PSP, 5 cases were initially misdiagnosed as adenocarcinoma and 1 as carcinoid preoperatively, while the remaining 73 cases were correctly diagnosed. 84.8% (67/79) of the PSP presented with well-defined, homogeneous, solitary solid tumors on chest imaging. Morphologically, 26.6% (21/79) of the PSP mainly showed a single histological component, 67.1% (53/79) contained two histological components, and 6.3% (5/79) contained three histological components. There were no cases containing all four histological components simultaneously. The tumor was composed of cuboidal cells on the surface and round cells in the stroma and lacked significant cytological atypia and mitotic figures. Some cases exhibited variations in histology and cellular morphology, such as glandular spaces (58.2%, 46/79), sclerotic papillae (46.8%, 37/79), hypercellularity (16.5%, 13/79), and cytological atypia (24.1%, 19/79). Immunophenotyping indicated that both tumor cell types expressed TTF1, EMA and β-catenin, while surface cells expressed pan-cytokeratin and Napsin A, and stromal cells expressed vimentin. In some cases, ER and PR were also expressed.Conclusions:When diagnosing PSP in needle biopsy specimens, the key to avoiding misdiagnosis is recognizing the presence of dual-cell populations within the tumor. The useful clues include presence of cellular papillae, mild cellular atypia, morphological diversity, interstitial foam-like cell aggregates, and prominent background hemorrhage and sclerosis. The characteristic immunophenotype and middle-aged female predilection are also helpful for the diagnosis of PSP.
3.Identification of Arisaematis Rhizoma and Pinelliae Rhizoma by PCR-RFLP
Jiancai XIAO ; Binbin YAN ; Jian YANG ; Jiahui SUN ; Tielin WANG ; Xiufu WAN ; Kai SUN ; Chao JIANG ; Yan ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(6):194-201
ObjectiveIn recent years, with the sharp decline of wild resources in Arisaematis Rhizoma and Pinelliae Rhizoma and the immaturity of medicinal cultivation technology, their adulterants have appeared frequently in the market, and the main identifying characteristics have mostly disappeared in the circulation of medicinal materials. Therefore, there is an urgent need to establish a molecular identification method that can quickly and effectively identify the specificity of Arisaematis Rhizoma and Pinelliae Rhizoma. MethodAfter comparison of the rbcL sequences of Arisaematis Rhizoma,Pinelliae Rhizoma, and their adulterants, the specific enzyme cleavage sites Hae Ⅲ and Dra Ⅰ of Arisaematis Rhizoma and Pinelliae Rhizoma, respectively, were selected and identified by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The main system conditions of PCR-RFLP reaction were established and optimized, and their durability and the ability to detect genuine, adulterants, and mixed counterfeits were investigated. ResultThe PCR-RFLP identification method of Arisaematis Rhizoma and Pinelliae Rhizoma was established. After specific primer amplification, Arisaematis Rhizoma and Pinelliae Rhizoma could be digested by Hae Ⅲ and Dra Ⅰ-restricted endonucleases respectively, at annealing temperature of 54 ℃, the number of cycles of 35, and the amount of DNA template of 3-30 ng, producing two fragments or small cut fragments with a single band between 100-250 bp, whereas the mixed counterfeits were not cleaved and both showed a band at 250 bp. The method is highly accurate in identifying adulterants and mixed counterfeits of Arisaematis Rhizoma or Pinelliae Rhizoma. ConclusionThe PCR-RFLP method developed in this study allows for the rapid identification of Arisaematis Rhizoma and Pinelliae Rhizoma.
4.Effect of Light and Temperature on Photosynthetic Fluorescence Parameters and Volatile Oil Content of Atractylodes lancea
Meiyu CAO ; Kai SUN ; Binbin YAN ; Sheng WANG ; Tielin WANG ; Yan ZHANG ; Lanping GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(20):108-115
5.Determination of formononetin and harmful elements of heavy metals in Jinji Pill
Tao CHENG ; Yingchao WANG ; Ke SHU ; Tielin WANG ; Ling CHANG
International Journal of Traditional Chinese Medicine 2021;43(10):1006-1010
Objective:To use the high performance liquid chromatography method to determine the content of formononetin in Jinji Pills and by using atomic absorption spectrophotometry,method to determine the harmful elements of heavy metal in Jinji Pills in orer to provide the scientific foundation for improving its quality standards and safety evaluation. Methods:Use Waters XBridge? C18 column (4.6 mm × 250 mm, 5 μm), set mobile phase at acetonitrile-1% phosphoric acid solution (27:73), flow rate 1.0 ml/min, column temperature 30 ℃, detection wavelength 249 nm, column temperature 30 ℃; Lead (Pb) and cadmium (Cd) was detected by graphite furnace method; arsenic (As) was detected by cold steam series graphite furnace method; copper (Cu) was detected by flame method; mercury (Hg) was detected by cold steam method.Results:The formononetin had a good linear relationship between 0.02-2.01 μg, the recovery rate was 98.5%, RSD was 1.53%. Lead (Pb) recovery rate was 103.6%, cadmium (Cd) recovery rate was 95.7%, arsenic (As) recovery rate was 92.4%, mercury (Hg) recovery rate was 104.9%, copper (Cu) recovery rate was 112.5%. Conclusion:This method is of accuracy, specificity, high sensitivity and good reproducibility, which could provide strong evidence for quality improvement and safety use of Jinji pill.
6.Relationship between TRX1 and JAB1 in acute myelogenous leukemia
Lingyun HUI ; Qiuju SHEN ; Ting WANG ; Shan MENG ; Chenyang LU ; Tielin YANG ; Wanggang ZHANG ; Fuling ZHOU
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(1):62-65,70
ABSTRACT:Objective To detect the expressions of thioredoxin (TRX1)and c-jun-activation-domain binding protein-1 (JAB1)in patients with acute myelogenous leukemia (AML)and healthy controls,and measure the TRX1 level in AML patients at different stages for evaluating its clinical significance.Methods The expressions of TRX1 and JAB1 in leukemia samples were analyzed by RT-PCR and Western blot at mRNA and protein levels, respectively.The correlation between TRX1 and JAB1,and the relationship between the gene expression and peripheral blood leukocytes count were also analyzed.Furthermore,serum TRX1 was measured by ELISA.Results TRX1 and JAB1 expressions at both mRNA and protein levels were obviously upregulated in leukemia patients (P<0.05). TRX1 was positively related to JAB1 in both newly diagnosed and recurrent AML patients.And high levels of TRX1 and JAB1 expressions were associated with white blood cell (WBC)counts in AML patients (P<0.05).Moreover, abundance of TRX1 in serum was significantly greater in AML patients,especially in the patients with recurrent AML,than in healthy donors (P<0.05).Conclusion There is a positive correlation between the expressions of TRX1 and JAB1 ,which is closely related to the occurrence and progression of AML.
7.One case of granular cell tumor of the breast
Fan FENG ; Tielin WANG ; Guosheng REN ; Fan LI
Chinese Journal of Endocrine Surgery 2017;11(2):161-163
8.Effect of methylprednisolone pre-intervention on expressions of heat shock protein 27 and tumor necrosis factor alpha in spinal cord cells following ischemia-reperfusion injury in rats
Lei XIA ; Tielin YIN ; Lin DU ; Ruonan ZHOU ; Yisheng WANG
Chinese Journal of Orthopaedic Trauma 2009;11(6):555-558
Objective To observe the effect of methylprednisolone (MP) pre-intervention on ex-pressions of heat shock protein 27 (HSP27) and tumor necrosis factor alpha (TNF-α) in cells in rat spinal cord following ischemia-reperfusion injury. Methods One hundred and fifty male Sprague-Dawley (SD) rats were randomly divided into 3 equal groups: group A (control) in which the abdominal aorta was exposed without any treatment, group B in which the abdominal aorta was clipped for 30 minutes before reperfusion for 3 bours to establish a model of ischemia- reperfusion injury, and group C in which intravenous MP injection was conducted 30 minutes before the establishment of the ischemia-reperfusion injury model. Three hours later the spinal cords were harvested. Pathological changes of spinal cord cells were observed with HE staining and expressions of HSP27 and TNF-α in spinal cord cells were observed with immunohistochemical staining. The motor function of hind-limbs before was evaluated before sample harvest. The data were analyzed with SPSS software. Results There were significant differences between groups A and B in the expressions of TNF-α and HSP27. Compared with group B, the expression of TNF-α decreased and HSP27 increased in group C, with statistically significant differences between the 2 groups. The motor function score of hind-limbs decreased in group B but improved in group C. Conclusions Since MP can decrease the expression of TNF-α and up-regulate the expression of HSP27, it has a potency of neuro-protection. Spinal cord ischemia-reperfusion injury can be avoided or decreased after MP pre-intervention.
9.Primitive trigeminal artery-cavernous sinus fistula and ICA-primitive trigeminal artery aneurysm:report of two cases and review of literatures
Yuanxing GUO ; Tielin LI ; Chuanzhi DUAN ; Qiujing WANG ; Qing HUANG
Chinese Journal of Cerebrovascular Diseases 2004;1(8):366-371
The authors described two cases with primitive trigeminal artery. Case 1 was a 32-year-old woman who suffered dizziness and a serious pulsatile intracranial bruit on the left ear, and sometimes associated with pulsatile intracranial bearing-pain on the left temporal side six months before she was admitted to the hospital. She also suffered from obvious diplopia on left lateral gaze for the last 5 months. She had suffered no recent trauma. Magnetic resonance imaging(MRI) demonstrated a suspected intracranial aneurysm located in left cavernous sinus. Digital subtraction angiography (DSA) was performed and a primitive trigeminal artery-cavernous sinus fistula in left side was found. Intraluminal occlusion of the fistula was successfully performed immediately after angiography using 6 Guglielmi detachable coils (GDC), and the patient was cured finally. Case 2 was a 28-year-old woman who suffered a serious intermittent cephalodynia associated with soreness on the left body two years before she was admitted to the hospital. She had suffered no recent trauma. Magnetic resonance angiography(MRA) demonstrated a suspected intracavernous aneurysm of the right internal carotid artery, Digital subtraction angiography (DSA) was performed. Right internal carotid angiography showed a primitive trigeminal artery (PTA) run between the cavernous segment of the internal carotid artery and the distal portion of the basilar artery. On initiation of PTA of R-ICA a small wide-necked saccular aneurysm was incidentally visualized. The aneurysm was successfully embolized after angiography using 2 Stent (Neuroform, 4.5mmm × 20mmm)-assisted detachable coils (Matrix), the ICA and PTA were preserved, and the patient was cured finally.
10.Three-dimensional CT angiography with volume rendering for the dignosis of multiple intracranial aneurysms
Bing FANG ; Tielin LI ; Jianmin ZHANG ; Chuanzhi DUAN ; Qiujing WANG ; Qingping ZAO
Chinese Journal of Cerebrovascular Diseases 2004;1(1):36-42
Objective:To evaluate the importance of 3D-CTA with volume rendering for the diagnosis of multiple intracranial aneurysms. Methods: Axial source images were obtained by helical CT scanning and reconstruction of 3D-CTA images was done by volume rendering technique in conjunction with multiplanar reformation. Results: In the past one year,there were 10 patients diagnosed as having multiple intracranial aneurysms by 3D-CTA and altogether 24 aneurysms were visualized,including 10 small aneurysms(≤5mm.Three dimensional CT angiography with volume rendering demonstrated aneurysms very well and provided useful information concerning the site,shape,size and spatial relationship with the surrounding vessels and bone anatomy. Conclusion: Three-dimensional CT angiography with volume rendering is a quick,reliable,and relatively noninvasive method for diagnosing multiple intracranial aneurysms.It delineates detailed aneurysmal morphology,and provides useful information for planning microsurgical approaches.

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