1.Mechanism of Xiangmei Pills in treating ulcerative colitis based on UHPLC-Q-Orbitrap HRMS and 16S rDNA sequencing of intestinal flora.
Ya-Fang HOU ; Rui-Sheng WANG ; Zhen-Ling ZHANG ; Wen-Wen CAO ; Meng ZHAO ; Ya-Hong ZHAO
China Journal of Chinese Materia Medica 2025;50(4):882-895
The efficacy of Xiangmei Pills on rats with ulcerative colitis(UC) was investigated by characterizing the spectrum of the active chemical components of Xiangmei Pills. Rapid identification and classification of the main chemical components were performed,and the therapeutic effects of Xiangmei Pills on the proteins and intestinal flora of UC rats were analyzed to explore the mechanism of its action in treating UC. Fifty SD rats were acclimatized to feeding for 3 d and randomly divided into blank group, model group,mesalazine group(0. 4 g·kg~(-1)), low-dose group of Xiangmei Pills(1. 89 g·kg~(-1)), and high-dose group of Xiangmei Pills(5. 67 g·kg~(-1)), with 10 rats in each group. 5% dextrose sodium sulfate(DSS) was given by gavage to induce the male SD rat model with UC,and the corresponding medicinal solution was given by gavage after 10 days, respectively. The therapeutic effect of Xiangmei Pills on rats with UC was evaluated according to body mass, disease activity index(DAI), and hematoxylin-eosin(HE) staining, and the histopathological changes in the colon were observed. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) technique was used to rapidly and accurately identify the main chemical constituents of Xiangmei Pills. Immunohistochemistry was used to detect the expression of aryl hydrocarbon receptor(AhR),interferon-γ(IFN-γ), mucin-2(MUC-2), and cytochrome P450 1A1(CYP1A1) in colon tissue. Interleukin-22(IL-22) expression in colon tissue was detected by immunofluorescence. The 16S r DNA high-throughput sequencing technique was used to study the modulatory effects of Xiangmei Pills on the intestinal flora structure of rats with UC. Pharmacodynamic results showed that compared with that of the blank group, the colon tissue of the model group was congested, and ulcers were visible in the mucosa; compared with that in the model group, the histopathology of the colon of the rats with UC in the groups of Xiangmei Pills were improved, with scattered ulcers and reduced inflammatory cell infiltration. Chemical analysis showed that a total of 45 components were identified by mass spectrometry information, including 15 phenolic acids, 8 coumarins, 15 organic acids, 3 amino acids, 2 flavonoids, and 2 other components. Compared with those in the blank group, the levels of Ah R, CYP1A1, MUC-2, and IL-22 proteins in the colon tissue of rats in the model group were significantly decreased, and the level of IFN-γ protein was significantly increased; the intestinal flora of rats in the model group was disorganized, with a decrease in the abundance of the flora; the relative abundance of Bacteroidetes,unclassified genera of Ascomycetes, Prevotella of the Prevotella family, and Prevotella decreased significantly, and that of Firmicutes decreased, but the difference was not statistically significant. The relative abundance of Bacteroidetes, Bifidobacterium, and Lactobacillus increased significantly. Compared with those of the model group, the levels of Ah R, CYP1A1, MUC-2, and IL-22proteins in the colonic tissue of the groups of Xiangmei Pills were significantly higher, and the levels of IFN-γ proteins were significantly lower. The recovery of the intestinal flora was accelerated, and the diversity of the intestinal flora was significantly increased. The relative abundance of Bacteroidetes was significantly increased, and that of unclassified genera of Ascomycetes,Lactobacillus, Prevotella of the Prevotella family, and Prevotella was significantly increased. The relative abundance of Bacteroidetes and Bifidobacterium was significantly decreased. This study demonstrated that Xiangmei Pills can effectively treat UC, mainly through the phenolic acid and organic acid components to stimulate the intestinal barrier, regulate protein expression and the relative abundance and diversity of intestinal flora, and play a role in the treatment of UC.
Animals
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Colitis, Ulcerative/metabolism*
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Drugs, Chinese Herbal/chemistry*
;
Rats, Sprague-Dawley
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Male
;
Rats
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Gastrointestinal Microbiome/genetics*
;
Chromatography, High Pressure Liquid
;
Humans
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Mass Spectrometry
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RNA, Ribosomal, 16S/genetics*
;
Bacteria/drug effects*
2.Clinical Features, Prognostic Analysis and Predictive Model Construction of Central Nervous System Invasion in Peripheral T-Cell Lymphoma.
Ya-Ting MA ; Yan-Fang CHEN ; Zhi-Yuan ZHOU ; Lei ZHANG ; Xin LI ; Xin-Hua WANG ; Xiao-Rui FU ; Zhen-Chang SUN ; Yu CHANG ; Fei-Fei NAN ; Ling LI ; Ming-Zhi ZHANG
Journal of Experimental Hematology 2025;33(3):760-768
OBJECTIVE:
To investigate the clinical features and prognosis of central nervous system (CNS) invasion in peripheral T-cell lymphoma (PTCL) and construct a risk prediction model for CNS invasion.
METHODS:
Clinical data of 395 patients with PTCL diagnosed and treated in the First Affiliated Hospital of Zhengzhou University from 1st January 2013 to 31st December 2022 were analyzed retrospectively.
RESULTS:
The median follow-up time of 395 PTCL patients was 24(1-143) months. There were 13 patients diagnosed CNS invasion, and the incidence was 3.3%. The risk of CNS invasion varied according to pathological subtype. The incidence of CNS invasion in patients with anaplastic large cell lymphoma (ALCL) was significantly higher than in patients with angioimmunoblastic T-cell lymphoma (AITL) (P <0.05). The median overall survival was significantly shorter in patients with CNS invasion than in those without CNS involvement, with a median survival time of 2.4(0.6-127) months after diagnosis of CNS invasion. The results of univariate and multivariate analysis showed that more than 1 extranodal involvement (HR=4.486, 95%CI : 1.166-17.264, P =0.029), ALCL subtype (HR=9.022, 95%CI : 2.289-35.557, P =0.002) and ECOG PS >1 (HR=15.890, 95%CI : 4.409-57.262, P <0.001) were independent risk factors for CNS invasion in PTCL patients. Each of these risk factors was assigned a value of 1 point and a new prediction model was constructed. It could stratify the patients into three distinct groups: low-risk group (0-1 point), intermediate-risk group (2 points) and high-risk group (3 points). The 1-year cumulative incidence of CNS invasion in the high-risk group was as high as 50.0%. Further evaluation of the model showed good discrimination and accuracy, and the consistency index was 0.913 (95%CI : 0.843-0.984).
CONCLUSION
The new model shows a precise risk assessment for CNS invasion prediction, while its specificity and sensitivity need further data validation.
Humans
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Lymphoma, T-Cell, Peripheral/pathology*
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Prognosis
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Retrospective Studies
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Central Nervous System Neoplasms/pathology*
;
Neoplasm Invasiveness
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Male
;
Female
;
Central Nervous System/pathology*
;
Middle Aged
;
Adult
3.Langerhans Cell Histiocytosis of Bone:Report of Eight Cases and Review of the Literature.
Ya BI ; Dan-Dan WU ; Fang-Ying YU ; Zhen-Hong FANG ; Bo HUANG
Acta Academiae Medicinae Sinicae 2025;47(2):325-332
Langerhans cell histiocytosis of bone is a rare tumor disease characterized by the large accumulation of CD1a+ and CD207+ dendritic cells in tissues of unknown cause.It mainly occurs in children aged 1-4 years old,with incidences of 4-6 per million in children and 1-2 per million in adults.Due to its low incidence,diverse clinical manifestations,and no obvious specificity of imaging manifestations,the definitive diagnosis and early treatment of this type of tumor are challenging.In this paper,we report 8 cases of Langerhans cell histiocytosis of bone and review the relevant literature published in the past five years to summarize the clinical characteristics,pathological features,diagnosis,treatment,and prognosis of this disease.
Humans
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Bone Diseases/therapy*
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Histiocytosis, Langerhans-Cell/therapy*
4.Clinical trial of Morinda officinalis oligosaccharides in the continuation treatment of adults with mild and moderate depression
Shu-Zhe ZHOU ; Zu-Cheng HAN ; Xiu-Zhen WANG ; Yan-Qing CHEN ; Ya-Ling HU ; Xue-Qin YU ; Bin-Hong WANG ; Guo-Zhen FAN ; Hong SANG ; Ying HAI ; Zhi-Jie JIA ; Zhan-Min WANG ; Yan WEI ; Jian-Guo ZHU ; Xue-Qin SONG ; Zhi-Dong LIU ; Li KUANG ; Hong-Ming WANG ; Feng TIAN ; Yu-Xin LI ; Ling ZHANG ; Hai LIN ; Bin WU ; Chao-Ying WANG ; Chang LIU ; Jia-Fan SUN ; Shao-Xiao YAN ; Jun LIU ; Shou-Fu XIE ; Mao-Sheng FANG ; Wei-Feng MI ; Hong-Yan ZHANG
The Chinese Journal of Clinical Pharmacology 2024;40(6):815-819
Objective To observe the efficacy and safety of Morinda officinalis oligosaccharides in the continuation treatment of mild and moderate depression.Methods An open,single-arm,multi-center design was adopted in our study.Adult patients with mild and moderate depression who had received acute treatment of Morinda officinalis oligosaccharides were enrolled and continue to receive Morinda officinalis oligosaccharides capsules for 24 weeks,the dose remained unchanged during continuation treatment.The remission rate,recurrence rate,recurrence time,and the change from baseline to endpoint of Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),Clinical Global Impression-Severity(CGI-S)and Arizona Sexual Experience Scale(ASEX)were evaluated.The incidence of treatment-related adverse events was reported.Results The scores of HAMD-17 at baseline and after treatment were 6.60±1.87 and 5.85±4.18,scores of HAMA were 6.36±3.02 and 4.93±3.09,scores of CGI-S were 1.49±0.56 and 1.29±0.81,scores of ASEX were 15.92±4.72 and 15.57±5.26,with significant difference(P<0.05).After continuation treatment,the remission rate was 54.59%(202 cases/370 cases),and the recurrence rate was 6.49%(24 cases/370 cases),the recurrence time was(64.67±42.47)days.The incidence of treatment-related adverse events was 15.35%(64 cases/417 cases).Conclusion Morinda officinalis oligosaccharides capsules can be effectively used for the continuation treatment of mild and moderate depression,and are well tolerated and safe.
5.Expressions of zinc homeostasis proteins,GPR39 and ANO1 mRNA in the sperm of asthenozoospermia patients and their clinical significance
Chun HE ; Fang-Fang DAI ; Jun-Sheng LIU ; Ya-Song GENG ; Jun-Xia ZHOU ; Yi-Zhen HU ; Bo ZHENG ; Shu-Song WANG
National Journal of Andrology 2024;30(1):18-25
Objective:To explore the expressions of zinc homeostasis-related proteins,G protein-coupled receptor 39(GPR39)and ANO1 mRNA in the sperm of patients with asthenozoospermia(AS),and analyze their correlation with sperm motility.Methods:We collected semen samples from 82 male subjects with PR+NP<40%,PR<32%and sperm concentration>15 × 106/ml(the AS group,n=40)or PR+NP≥40%,PR≥32%and sperm concentration>15 × 106/ml(the normal control group,n=42).We analyzed the routine semen parameters and measured the zinc content in the seminal plasma using the computer-assisted sperm analysis system,detected the expressions of zinc transporters(ZIP13,ZIP8 and ZNT10),metallothioneins(MT1G,MT1 and MTF),GPR39,and calcium-dependent chloride channel protein(ANO1)in the sperm by real-time quantitative PCR(RT qPCR),examined free zinc distribution in the sperm by laser confocal microscopy,and determined the expressions of GPR39 and MT1 proteins in the sperm by immunofluorescence staining,followed by Spearman rank correlation analysis of their correlation with semen parameters.Results:There was no statistically significant difference in the zinc concentration in the seminal plasma between the AS and normal control groups(P>0.05).Compared with the controls,the AS patients showed a significantly reduced free zinc level(P<0.05),relative expressions of MT1G,MTF,ZIP13,GPR39 and ANO1 mRNA(P<0.05),and that of the GPR39 protein in the AS group(P<0.05).No statistically significant differences were observed in the relative expression levels of ZIP8,ZNT10 and MT1 mRNA between the two groups(P>0.05).The relative expression levels of GPR39,ANO1,MT1G and MTF mRNA were positively correlated with sperm motility and the percentage of progressively motile sperm(P<0.05).Conclusion:The expressions of zinc homeostasis proteins(MT1G,MTF and ZIP13),GPR39 and ANO1 mRNA are downregulated in the sperm of asthenozoospermia pa-tients,and positively correlated with sperm motility.
6.Treatment Outcomes in COVID-19 Patients with Brucellosis: Case Series in Heilongjiang and Systematic Review of Literature.
Man Li YANG ; Jing Ya WANG ; Xing Yu ZONG ; Li GUAN ; Hui Zhen LI ; Yi Bai XIONG ; Yu Qin LIU ; Ting LI ; Xin Yu JI ; Xi Yu SHANG ; Hui Fang ZHANG ; Yang GUO ; Zhao Yuan GONG ; Lei ZHANG ; Lin TONG ; Ren Bo CHEN ; Yi Pin FAN ; Jin QIN ; Fang WANG ; Gang LIN ; Nan Nan SHI ; Yan Ping WANG ; Yan MA
Biomedical and Environmental Sciences 2023;36(10):930-939
OBJECTIVE:
Clinical characteristics and outcome in COVID-19 with brucellosis patients has not been well demonstrated, we tried to analyze clinical outcome in local and literature COVID-19 cases with brucellosis before and after recovery.
METHODS:
We retrospectively collected hospitalization data of comorbid patients and prospectively followed up after discharge in Heilongjiang Infectious Disease Hospital from January 15, 2020 to April 29, 2022. Demographics, epidemiological, clinical symptoms, radiological and laboratory data, treatment medicines and outcomes, and follow up were analyzed, and findings of a systematic review were demonstrated.
RESULTS:
A total of four COVID-19 with brucellosis patients were included. One patient had active brucellosis before covid and 3 patients had nonactive brucellosis before brucellosis. The median age was 54.5 years, and all were males (100.0%). Two cases (50.0%) were moderate, and one was mild and asymptomatic, respectively. Three cases (75.0%) had at least one comorbidity (brucellosis excluded). All 4 patients were found in COVID-19 nucleic acid screening. Case C and D had only headache and fever on admission, respectively. Four cases were treated with Traditional Chinese medicine, western medicines for three cases, no adverse reaction occurred during hospitalization. All patients were cured and discharged. Moreover, one case (25.0%) had still active brucellosis without re-positive COVID-19, and other three cases (75.0%) have no symptoms of discomfort except one case fell fatigue and anxious during the follow-up period after recovery. Conducting the literature review, two similar cases have been reported in two case reports, and were both recovered, whereas, no data of follow up after recovery.
CONCLUSION
These cases indicate that COVID-19 patients with brucellosis had favorable outcome before and after recovery. More clinical studies should be conducted to confirm our findings.
Female
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Humans
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Male
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Middle Aged
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Brucellosis
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COVID-19
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Retrospective Studies
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SARS-CoV-2
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Treatment Outcome
;
Case Reports as Topic
7.3- to 24-month Follow-up on COVID-19 with Pulmonary Tuberculosis Survivors after Discharge: Results from a Prospective, Multicenter Study
Ya Jing WANG ; Yu Xing ZONG ; Hui Gui WU ; Lin Yuan QI ; Zhen Hui LI ; Yu Xin JI ; Lin TONG ; Lei ZHANG ; Bo Ming YANG ; Ye Pu YANG ; Ke Ji LI ; Rong Fu XIAO ; Song Lin ZHANG ; Hong Yun HU ; De Hong LIU ; Fang Shou XU ; Sheng SUN ; Wei WU ; Ya MAO ; Qing Min LI ; Hua Hao HOU ; Yuan Zhao GONG ; Yang GUO ; Wen Li JIAO ; Jin QIN ; Yi Ding WANG ; Fang WANG ; Li GUAN ; Gang LIN ; Yan MA ; Ping Yan WANG ; Nan Nan SHI
Biomedical and Environmental Sciences 2022;35(12):1091-1099
Objective Coronavirus disease 2019 (COVID-19) and tuberculosis (TB) are major public health and social issues worldwide. The long-term follow-up of COVID-19 with pulmonary TB (PTB) survivors after discharge is unclear. This study aimed to comprehensively describe clinical outcomes, including sequela and recurrence at 3, 12, and 24 months after discharge, among COVID-19 with PTB survivors. Methods From January 22, 2020 to May 6, 2022, with a follow-up by August 26, 2022, a prospective, multicenter follow-up study was conducted on COVID-19 with PTB survivors after discharge in 13hospitals from four provinces in China. Clinical outcomes, including sequela, recurrence of COVID-19, and PTB survivors, were collected via telephone and face-to-face interviews at 3, 12, and 24 months after discharge. Results Thirty-two COVID-19 with PTB survivors were included. The median age was 52 (45, 59) years, and 23 (71.9%) were men. Among them, nearly two-thirds (62.5%) of the survivors were moderate, three (9.4%) were severe, and more than half (59.4%) had at least one comorbidity (PTB excluded). The proportion of COVID-19 survivors with at least one sequela symptom decreased from 40.6% at 3 months to 15.8% at 24 months, with anxiety having a higher proportion over a follow-up. Cough and amnesia recovered at the 12-month follow-up, while anxiety, fatigue, and trouble sleeping remained after 24 months. Additionally, one (3.1%) case presented two recurrences of PTB and no re-positive COVID-19 during the follow-up period. Conclusion The proportion of long symptoms in COVID-19 with PTB survivors decreased over time, while nearly one in six still experience persistent symptoms with a higher proportion of anxiety. The recurrence of PTB and the psychological support of COVID-19 with PTB after discharge require more attention.
8.Quality Evaluation in Processing of Rehmanniae Radix Praeparata Processed with Amomi Fructus and Citri Reticulatae Pericarpium Based on "Color as Paint,Sweet as Maltose"
Qing-yao CHEN ; Xing LEI ; Ya-zhen GAO ; Jie XU ; Liang-feng WANG ; Ming YANG ; Xiao-ping WANG ; Fang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(4):154-162
ObjectiveTo study the correlation between the apparent color, comprehensive sweetness and the content of main components in the preparation of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium, so as to lay a foundation for revealing the processing principle of Rehmanniae Radix. MethodThe color of Rehmanniae Radix Praeparata sample powder was measured by automatic colorimeter, the contents of 14 active components in samples with different heating time points were determined by high performance liquid chromatography, including 7 glycosides of catalpol, rehmannia glycoside D, leonurine glycoside, 5-hydroxymethyl furfural, verbascoside, isoacteoside and hesperidin, and 7 carbohydrates of D-fructose, glucose, sucrose, melibiose, raffinose, manninotriose and stachyose), and the mobile phase was acetonitrile-water for gradient elution. The comprehensive sweetness difference of sample was calculated by the sweetness of saccharides, SPSS 21.0 was used to analyze the relationship between the color, comprehensive sweetness and the main component contents in the processing of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium, the quality comprehensive evaluation index of Rehmanniae Radix Praeparata by triangular area method was established. ResultDuring the processing, the color value of the powder increased, and the apparent color of the sample became darker. the content determination results showed that the content of glycosides decreased, monosaccharides and comprehensive sweetness increased with the increase of heating time. The results of correlation analysis showed that chromaticity value, comprehensive sweetness were significant negatively correlated with the content of iridoid glycosides (P<0.01), the chromaticity value was significant positively correlated with the contents of furaldehyde derivatives, phenylethanoid glycosides, flavonoids and comprehensive sweetness was significant positively correlated with the contents of furaldehyde derivatives, phenylethanoid glycosides (P<0.01), and the comprehensive sweetness was positively correlated with the content of flavonoids (P<0.05). After 52 h of processing, the comprehensive evaluation index of samples reached 0.99. ConclusionThe overall trend of cluster analysis of powder chromaticity value of Rehmanniae Radix Praeparata is basically consistent with that of naked eyes, the comprehensive quality evaluation of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium can be carried out by combining the three indexes of powder chromaticity value, comprehensive sweetness and glycosides content.
9.Effect of Different Steaming Processes on Variation of Active Ingredients in Notoginseng Radix et Rhizoma
Ya-zhen GAO ; Jun-bo ZOU ; Zhi-chao WANG ; Qing-yao CHEN ; Xing LEI ; Jie XU ; Ming YANG ; Fang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):147-154
ObjectiveTo study on the correlation between the apparent color, near-infrared spectroscopy and dynamic changes of index constituent content of samples during the processing of steamed Notoginseng Radix et Rhizoma, and to provide reference for the processing process optimization and quality control of this decoction piece. MethodSamples were dried and crushed by setting three steaming temperature gradients of 100-105, 114-118, 130-136 ℃, and sampled at steaming times of 1, 2, 4, 8, 12, 24 h, respectively. The effects of different steaming temperatures and times on the color and absorption of steamed Notoginseng Radix et Rhizoma at different infrared wavelengths were observed, and principal component analysis and cluster analysis were performed on the obtained data by R 4.1.0 and SPSS 21.0 software to observe the changes in color and infrared absorption characteristics of samples. High performance liquid chromatography (HPLC) was employed to determine the content changes of notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, 20(S)-ginsenoside Rg3 and 20(R)-ginsenoside Rg3 before and after steaming of Notoginseng Radix et Rhizoma, mobile phase was acetonitrile (A)-water (B) for gradient elution (0-30 min, 19%A; 30-60 min, 19%-44%A; 60-78 min, 44%-74%A; 78-80 min, 74%-100%A; 80-86 min, 100%A; 86-87 min, 100%-19%A; 87-95 min, 19%A) with the detection wavelength of 203 nm. ResultDuring the steaming process of Notoginseng Radix et Rhizoma, with the increase of steaming temperature and time, the b* (yellow-blue value), L* (brightness), △E* (comprehensive color difference value) of sample powder showed a decreasing trend, while the a* (red-green value) showed an increasing trend, and the color gradually deepened from gray to brown and dark black. There was no significant difference in the infrared absorption between raw and steamed Notoginseng Radix et Rhizoma sample powder in the low and medium wavelength bands, but significant difference in the infrared absorption of high band, especially in the band of 9 600-10 000 cm-1. HPLC showed that the contents of the original components (notoginsenoside R1, ginsenoside Rg1, Re and Rb1) decreased and 20(S)-ginsenoside Rg3 and 20(R)-ginsenoside Rg3 were newly produced after steaming of Notoginseng Radix et Rhizoma. The content proportion of these six index components reached the best when the steaming temperature at 130-136 ℃ and the steaming time of 1 h. ConclusionThe color and infrared absorption of samples are affected by the dynamic changes of chemical composition during the steaming process, and the composition change is the result of the joint influence of steaming temperature and time. In this paper, through multi-dimensional analysis of the apparent color indexes, the change pattern of near-infrared absorption characteristics and the index components of the samples, the different process parameters of steamed Notoginseng Radix et Rhizoma were evaluated holistically, and the quality transfer pattern of its processing process was initially revealed, which can provide scientific basis for processing optimization and quality evaluation of steamed Notoginseng Radix et Rhizoma.
10.Mechanism of Bushen Zhuyun Prescription on Improving Luteal Function of Brown Norway Rats Based on MAPKs Signaling Pathway
Xing-ran TANG ; Hui-fang ZHOU ; Hua FENG ; Yu-jie SHANG ; Yi-zhen YUAN ; Ya-xin DAI ; Yin-yin DING
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(12):78-85
ObjectiveTo investigate the possible mechanism of Bushen Zhuyun prescription (BSZYP) to reduce the level of ovarian apoptosis in Brown Norway (BN) rats with luteal phase deficiency (LPD). MethodFifty SPF female BN rats were randomly divided into a model group, a dydrogesterone group (0.002 g·kg-1), and a low (4.5 g·kg-1), a medium (9 g·kg-1), and a high-dose (18 g·kg-1) BSZYP groups, with ten rats in each group. The rats were administrated with corresponding drugs by gavage, once a day for three estrus cycle. Western blot was used to detected the protein expression levels of c-Jun NH2 terminal kinase (JNK), extracellular signal-regulated kinase (ERK), phosphorylated-ERK (p-ERK), phosphorylated-JNK (p-JNK), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated-p38 MAPK (p-p38 MAPK ), B-cell lymphoma (Bcl-2), and Bcl-2 associated X protein (Bax) in ovary. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of ERK, JNK, p38 MAPK, Bax, and Bcl-2 in ovary. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum progesterone (P) and estradiol (E2) levels. Hematoxylin-eosin (HE) staining was used to observe the ovarian tissue morphology. ResultCompared with the model group, the recovery of estrus cycle of rats in all BSZYP groups had statistical significance after 1-circle administration (P<0.05). The ovarian tissue morphology in the low-dose BSZYP group was improved, and that in the medium and high-dose BSZYP groups was significantly improved with clear follicle, less vesicular follicle and atretic follicle, and more granular layers. The number of luteum, especially the fresh luteum, in the medium and high-dose groups was increased with smooth edge and large volume. The mRNA expression level of Bcl-2 was increased in all-dose BSZYP groups, while the mRNA expression level of Bax was significantly decreased in all-dose BSZYP group (P<0.05, P<0.01). The mRNA expression levels of JNK and p38 MAPK were significantly decreased in the high-dose BSZYP group (P<0.01), and the mRNA expression levels of ERK were increased in the low and medium-dose BSZYP groups (P<0.05). The protein expression level of Bcl-2 was significantly increased in the medium and high-dose BSZYP groups (P<0.01), and the protein expression level of Bax was significantly decreased in the all-dose BSZYP groups (P<0.01). No significant difference was observed in the protein expressions of JNK, ERK, and p38 MAPK in the BSZYP groups. The protein expression levels of p-ERK in the ovarian tissues of rats were significantly inoreased in the medium and high-dose BSZYP group (P<0.01), p-JNK, and p-p38 MAPK in the ovarian tissues of rats were significantly decreased in the medium and high-dose BSZYP group (P<0.01). The level of E2 was increased in all-dose BSZYP groups (P<0.05, P<0.01), and the level of P in the medium-dose BSZYP group was increased (P<0.05). ConclusionBSZYP improved the serum sex hormones, restored the estrous cycle, reduced atretic follicle and vesiculation, and maintained luteal morphology and function of BN rats, so as to improve luteal function and treat luteal phase deficiency. The mechanism of BSZYP may be related to reduce the level of ovarian tissue apoptosis in BN rats by regulating MAPKs signaling pathway.

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