1.Effects of Electroacupuncture at "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) on Nociceptive Sensitization and PKC/TRPV1 Pathway in the Trigeminal Ganglion of Chronic Migraine Model Rats
Yixiang ZENG ; Runze TU ; Shucong ZHAO ; Yang YANG ; Haojia WEN ; Zhuozhong HE ; Shengli ZHOU ; Lei TAN ; Ke HE ; Lei FU
Journal of Traditional Chinese Medicine 2025;66(3):283-289
ObjectiveTo explore the possible mechanisms of electroacupuncture at Fengchi (GB 20), Waiguan (TE 5), and Yanglingquan (GB 34) in treating chronic migraine from the perspective of nociceptive sensitization. MethodsForty SPF-grade SD rats were randomly divided into blank group, model group, electroacupuncture group, electroacupuncture + agonist group, and inhibitor group, with 8 rats in each group. Except for the blank group, rats were injected intraperitoneally with nitroglycerin to establish a chronic migraine rat model. After successful modeling, the electroacupuncture group received electroacupuncture at bilateral "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) for 30 minutes each session. The electroacupuncture + agonist group received the same electroacupuncture treatment and additional injection of protein kinase C (PKC) agonist Phorbol 12-myristate 13-acetate (1.0 ng/μl, 25 μl) via the infraorbital foramen. The inhibitor group received PKC inhibitor Chelerythrine Chloride (1.0 ng/μl, 10 μl) via the infraorbital foramen. The blank group, model group, and inhibitor group underwent restraint for 30 minutes without other interventions. All groups were continuously intervened for 5 days. After the intervention, the nociceptive thresholds (mechanical and thermal pain) of the periorbital area and hind paw were measured. The expression levels of transient receptor potential vanillic acid subtype 1 (TRPV1), phosphorylated TRPV1 (p-TRPV1), PKC proteins, Trpv1, Pkc mRNA, and the average fluorescence intensity of transient receptor potential vanillic acid subtype 1 (TRPV1) and PKC in the trigeminal ganglion were detected using Western Blot, real-time fluorescence quantitative PCR, and immunofluorescence methods. ResultsCompared with the blank group, the mechanical and thermal pain thresholds of the periorbital area and hind paw were reduced in the model group, and the protein levels of TRPV1, PKC, p-TRPV1, as well as the mRNA expression of Trpv1 and Pkc, and the average fluorescence intensity of TRPV1 and PKC in the trigeminal ganglion significantly increased (P<0.05 or P<0.01). Compared with the model group, the electroacupuncture group exhibited increased mechanical and thermal pain thresholds in the periorbital and hind paw areas, and decreased protein levels of TRPV1, PKC, p-TRPV1, mRNA expression of Trpv1 and Pkc, and average fluorescence intensity of TRPV1. In the electroacupuncture + agonist group, the average fluorescence intensity of TRPV1 in the trigeminal ganglion decreased. The inhibitor group exhibited increased mechanical pain thresholds in the periorbital area and thermal pain thresholds in the hind paw, along with decreased protein levels of TRPV1, PKC, p-TRPV1, and the average fluorescence intensity of TRPV1 and PKC (P<0.05 or P<0.01). Compared with the electroacupuncture group, the electroacupuncture + agonist group showed an increase in the protein levels of TRPV1, PKC, p-TRPV1, and the mRNA expression of Trpv1 (P<0.05 or P<0.01). ConclusionElectroacupuncture at the "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) acupoints can increase the mechanical and thermal pain thresholds in chronic migraine rats and alleviate nociceptive sensitization. The mechanism may be related to the inhibition of PKC/TRPV1 pathway.
2.Processing technology of calcined Magnetitum based on concept of QbD and its XRD characteristic spectra.
De-Wen ZENG ; Jing-Wei ZHOU ; Tian-Xing HE ; Yu-Mei CHEN ; Huan-Huan XU ; Jian FENG ; Yue YANG ; Xin CHEN ; Jia-Liang ZOU ; Lin CHEN ; Hong-Ping CHEN ; Shi-Lin CHEN ; Yuan HU ; You-Ping LIU
China Journal of Chinese Materia Medica 2025;50(9):2391-2403
Guided by the concept of quality by design(QbD), this study optimizes the calcination and quenching process of calcined Magnetitum and establishes the XRD characteristic spectra of calcined Magnetitum, providing a scientific basis for the formulation of quality standards. Based on the processing methods and quality requirements of Magnetitum in the Chinese Pharmacopoeia, the critical process parameters(CPPs) identified were calcination temperature, calcination time, particle size, laying thickness, and the number of vinegar quenching cycles. The critical quality attributes(CQAs) included Fe mass fraction, Fe~(2+) dissolution, and surface color. The weight coefficients were determined by combining Analytic Hierarchy Process(AHP) and the criteria importance though intercrieria correlation(CRITIC) method, and the calcination process was optimized using orthogonal experimentation. Surface color was selected as a CQA, and based on the principle of color value, the surface color of calcined Magnetitum was objectively quantified. The vinegar quenching process was then optimized to determine the best processing conditions. X-ray diffraction(XRD) was used to establish the characteristic spectra of calcined Magnetitum, and methods such as similarity evaluation, cluster analysis, and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to evaluate the quality of the spectra. The optimized calcined Magnetitum preparation process was found to be calcination at 750 ℃ for 1 h, with a laying thickness of 4 cm, a particle size of 0.4-0.8 cm, and one vinegar quenching cycle(Magnetitum-vinegar ratio 10∶3), which was stable and feasible. The XRD characteristic spectra analysis method, featuring 9 common peaks as fingerprint information, was established. The average correlation coefficient ranged from 0.839 5-0.988 1, and the average angle cosine ranged from 0.914 4 to 0.995 6, indicating good similarity. Cluster analysis results showed that Magnetitum and calcined Magnetitum could be grouped together, with similar compositions. OPLS-DA discriminant analysis identified three key characteristic peaks, with Fe_2O_3 being the distinguishing component between the two. The final optimized processing method is stable and feasible, and the XRD characteristic spectra of calcined Magnetitum was initially established, providing a reference for subsequent quality control and the formulation of quality standards for calcined Magnetitum.
X-Ray Diffraction/methods*
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Drugs, Chinese Herbal/chemistry*
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Quality Control
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Particle Size
3.Mediating effect of sleep duration between depression symptoms and myopia in middle school students.
Wei DU ; Xu-Xiang YANG ; Ru-Shuang ZENG ; Chun-Yao ZHAO ; Zhi-Peng XIANG ; Yuan-Chun LI ; Jie-Song WANG ; Xiao-Hong SU ; Xiao LU ; Yu LI ; Jing WEN ; Dang HAN ; Qun DU ; Jia HE
Chinese Journal of Contemporary Pediatrics 2025;27(3):359-365
OBJECTIVES:
To explore the mediating role of sleep duration in the relationship between depression symptoms and myopia among middle school students.
METHODS:
This study was a cross-sectional research conducted using a stratified cluster random sampling method. A total of 1 728 middle school students were selected from two junior high schools and two senior high schools in certain urban areas and farms of the Xinjiang Production and Construction Corps. Questionnaire surveys and vision tests were conducted among the students. Spearman analysis was used to analyze the correlation between depression symptoms, sleep duration, and myopia. The Bootstrap method was employed to investigate the mediating effect of sleep duration between depression symptoms and myopia.
RESULTS:
The prevalence of myopia in the overall population was 74.02% (1 279/1 728), with an average sleep duration of (7.6±1.0) hours. The rate of insufficient sleep was 83.62% (1 445/1 728), and the proportion of students exhibiting depression symptoms was 25.29% (437/1 728). Correlation analysis showed significant negative correlations between visual acuity in both eyes and sleep duration with depressive emotions as measured by the Center for Epidemiologic Studies Depression Scale (with correlation coefficients of -0.064, -0.084, and -0.199 respectively; P<0.01), as well as with somatic symptoms and activities (with correlation coefficients of -0.104, -0.124, and -0.233 respectively; P<0.01) and interpersonal relationships (with correlation coefficients of -0.052, -0.059, and -0.071 respectively; P<0.05). The correlation coefficients for left and right eye visual acuity and sleep duration were 0.206 and 0.211 respectively (P<0.001). Sleep duration exhibited a mediating effect between depression symptoms and myopia (indirect effect=0.056, 95%CI: 0.029-0.088), with the mediating effect value for females (indirect effect=0.066, 95%CI: 0.024-0.119) being higher than that for males (indirect effect=0.042, 95%CI: 0.011-0.081).
CONCLUSIONS
Sleep duration serves as a partial mediator between depression symptoms and myopia in middle school students.
Humans
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Myopia/etiology*
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Male
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Female
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Depression/physiopathology*
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Cross-Sectional Studies
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Sleep
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Adolescent
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Students
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Child
;
Time Factors
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Sleep Duration
4.Analysis of Gene Mutations Distribution and Enzyme Activity of G6PD Deficiency in Newborns in Guilin Region.
Dong-Mei YANG ; Guang-Li WANG ; Dong-Lang YU ; Dan ZENG ; Hai-Qing ZHENG ; Wen-Jun TANG ; Qiao FENG ; Kai LI ; Chun-Jiang ZHU
Journal of Experimental Hematology 2025;33(5):1405-1411
OBJECTIVE:
To analyze the distribution characteristics of glucose-6-phosphate-dehydrogenase (G6PD) mutations and their enzyme activity in newborns patients with G6PD deficiency in Guilin region.
METHODS:
From July 2022 to July 2024, umbilical cord blood samples from 4 554 newborns in Guilin were analyzed for G6PD mutations using fluorescence PCR melting curve analysis. Enzyme activity was detected in 4 467 cases using the rate assay.
RESULTS:
Among 4 467 newborns who underwent G6PD activity testing, 162 newborns (3.63%) were identified as G6PD-deficient, including 142 males (6.04%) and 20 females (0.94%), the prevalence of G6PD deficiency was significantly higher in males than in females (P < 0.001). Genetic analysis of 4 554 newborns detected G6PD mutations in 410 cases (9%), including 171 males (7.13%) and 239 females (11.09%), with a significantly higher mutation detection rate in females than in males (P < 0.001). A total of nine single mutations and four compound heterozygous mutations were identified. The most common mutations were c.1388G>A (33.66%), c.1376G>T (23.66%) and c.95A>G (16.34%). Among newborns who underwent both enzyme activity and genetic mutation testing, males with G6PD mutations had significantly lower enzyme activity than that of females with G6PD mutations(P < 0.001). Specifically, among newborns carrying the mutations c.1388G>A, c.1376G>T, c.95A>G, c.1024C>T or c.871G>A, males consistently exhibited lower enzymatic activity than females with the same mutations (P < 0.001). Furthermore, in male G6PD-deficient newborns, the enzyme activity levels in those carrying c.1388G>A, c.1376G>T, c.95A>G, c.1024C>T, or c.871G>A were lower than those in both the control group and the c.519C>T group (P < 0.05).
CONCLUSION
This study provides a comprehensive profile of G6PD deficiency incidence and mutation spectrum in the Guilin region. By analyzing enzyme activity and genetic mutation results, this study provides insights into potential intervention strategies and personalized management approaches for the prevention and treatment of neonatal G6PD deficiency in the region.
Humans
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Infant, Newborn
;
Glucosephosphate Dehydrogenase Deficiency/epidemiology*
;
Glucosephosphate Dehydrogenase/genetics*
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Female
;
Male
;
Mutation
;
China/epidemiology*
5.Integrated imaging and clinical features of glottic squamous cell carcinoma of the larynx: pathological association and prognosis assessment.
Yuqiao ZHANG ; Wulin WEN ; Fengxia YANG ; Dongke MA ; Xueliang SHEN ; Ningyu FENG ; Xixi LI ; Zhiling ZENG ; Zhipeng MI ; Xiyuan YAN ; Ruixia MA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(8):709-716
Objective:To explore the clinical, imaging, and pathological features of glottic squamous cell carcinoma of the larynx and their relationship with prognosis. Methods:A retrospective analysis was conducted on the clinical, imaging, and pathological data of 130 patients with glottic squamous cell carcinoma of the larynx who were treated at the First People's Hospital of Yinchuan and the General Hospital of Ningxia Medical University from January 2018 to March 2023. Imaging examinations (CT and MRI) were used to evaluate the lesion boundary clarity, density, enhancement nature, and enhancement degree. Postoperative pathological examination was used to determine the pathological nature, immunohistochemistry, etc. Statistical methods such as χ² test, Spearman correlation analysis, multivariate logistic regression analysis, and Kaplan-Meier method were used to analyze the data. Results:Among the 130 patients, 127 were male and 3 were female, with an average age of (61.92±9.595) years. There was a correlation between clinical, imaging, and pathological features. Multivariate analysis showed that heterogeneous MRI density (OR=12.414;P=0.019) and squamous cell carcinoma as a subtype were correlated. The initial symptom of non-hoarseness (HR=6.045;P=0.010) and unclear MRI boundary (HR=12.559; P=0.029) were independent risk factors for poor prognosis in patients with glottic squamous cell carcinoma of the larynx. Conclusion:There is a correlation between the clinical, imaging, and pathological features of patients with glottic squamous cell carcinoma of the larynx, and they can affect prognosis. The initial symptom of non-hoarseness and unclear MRI boundary of the tumor are independent risk factors for poor prognosis.
Humans
;
Laryngeal Neoplasms/diagnosis*
;
Prognosis
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Male
;
Female
;
Retrospective Studies
;
Middle Aged
;
Carcinoma, Squamous Cell/diagnosis*
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Magnetic Resonance Imaging
;
Glottis/pathology*
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Tomography, X-Ray Computed
;
Aged
6.A Sensitive Lateral Flow Immunoassay for Detection of Interleukin-6 Using Carbon Dots-Mesoporous Silica Nanocomposite Fluorescent Probes
Yue-Qian YANG ; Peng-Yue WANG ; Jia-Qi REN ; Xiao PAN ; Feng-Hua TAN ; Yu-Jie MA ; Cong-Ying WEN ; Jing-Bin ZENG
Chinese Journal of Analytical Chemistry 2025;53(9):1467-1475
In this study,a sensitive lateral flow immunoassay(LFIA)platform based on carbon dots-mesoporous silica nanocomposite(CD-MSNs)fluorescent probes was constructed for high-performance detection of inflammatory marker interleukin-6(IL-6).Green fluorescent carbon dots(CDs)were prepared by hydrothermal method with 3,9-perylenic acid and 3-aminopropyltriethoxysilane(APTES)as raw materials,and highly fluorescent CD-MSNs composites were then constructed by encapsulating the prepared CDs in mesoporous silica nanoparticles(MSNs).Fluorescent probes were prepared by covalent coupling of CD-MSNs with IL-6 antibody.Fluorescent immunochromatographic test strips were constructed by spraying IL-6 capture antibody and goat anti-mouse IgG on nitrocellulose membrane as detection line(T-line)and quality control line(C-line),respectively.The fluorescence immunoassay analyzer was used to quantitatively detect the fluorescence intensity of T-line,and the experimental results showed that the LFIA platform based on this probe had a good linear relationship in IL-6 concentration range of 102-106 pg/mL,and the detection limit was 64 pg/mL,which was two orders of magnitude more sensitive than that of the traditional colloidal gold test strips.This method effectively solved the issue of insufficient sensitivity of traditional LFIA technique,and provided a rapid and highly sensitive detection method for early diagnosis of inflammatory diseases.
7.The Role of Ferroptosis in Hepatocyte Injury Induced by α-Amanitin
Hao-Wei WANG ; Xiao-Xing ZHANG ; Gen-Meng YANG ; Shang-Wen WANG ; Xiao-Feng ZENG
Journal of Forensic Medicine 2025;41(2):152-159
Objective To explore whether ferroptosis is involved in α-amanitin-induced hepatocyte in-jury by detecting iron deposition in mice liver tissues,oxidative stress indicators in hepatocytes and L-02 cells,and expressions of ferroptosis-related proteins after α-amanitin exposure.Methods The poi-soning models of α-amanitin C57BL/6J mice and L-02 cell were established.The Lillie ferrous iron staining and Prussian blue staining were used to detect iron deposition;the kits were applied to detect the levels of superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),and glutathione(GSH).Western blotting was performed to analyze expressions of p53,solute carrier family 7 member 11(SLC7A11),and glutathione peroxidase 4(GPX4).Results Compared with the control group,after α-amanitin exposure,positive cell rates of Fe2+and Fe3+in mice liver tissues increased significantly.In the liver tissues of medium(0.35 mg/kg)and high(0.45 mg/kg)dose groups and L-02 cells treated with 1 μmol/L α-amanitin,the level of GSH decreased,the level of MDA increased,and the activities of SOD and CAT decreased significantly.In addition,α-amanitin upregulated the expression of p53 in a concentration-and time-dependent manner and inhibited the expressions of SLC7A11 and GPX4.Con-clusion Ferroptosis plays an important role in α-amanitin-induced hepatocyte injury.Abnormalities of ferroptosis-related indicators can provide references for the forensic identification of α-amanitin poisoning.
8.Effects of electroacupuncture at acupoints of Shaoyang meridian on peripheral TRPV1 and CGRP in rats with chronic migraine
Runze TU ; Yixiang ZENG ; Shengli ZHOU ; Haojia WEN ; Yang YANG ; Zhuozhong HE ; Weiai LIU ; Ke HE ; Lei FU
International Journal of Traditional Chinese Medicine 2025;47(4):483-487
Objective:To observe the effects of electroacupuncture (EA) at acupoints of Shaoyang meridian on mechanical and thermal pain threshold, TRPV1, CGRP mRNA and TRPV1, p-TRPV1, CGRP protein expressions in TG of CM rats; To explore its mechanism.Methods:Totally 24 SD rats were divided into blank group, model group and EA group according to random number table method, with 8 rats in each group. The blank group did not do any treatment; the model group and the EA group were given intraperitoneal injection of nitroglycerin (10 mg/kg) on the 1st, 3rd, 5th, 7th and 9th day, once a day, for 5 times to prepare migraine rat model; after modeling, the model group was given binding and restraint for 30 min, and the EA group was given binding and restraint for 30 min treatment; the changes of basic mechanical and thermal pain thresholds of rats before and after modeling and intervention were observed, the mRNA expression levels of TRPV1 and CGRP in TG rats were detected by qPCR, and the protein expression levels of TRPV1, p-TRPV1 and CGRP in TG rats were detected by Western blot method.Results:After intervention, compared with the model group, the periorbital and plantar mechanical pain thresholds and plantar thermal pain thresholds in EA group increased ( P<0.05), the TRPV1 and CGRP mRNA expressions in trigeminal ganglion decreased ( P<0.01), the protein expressions of TRPV1, p-TRPV1 and CGRP decreased ( P<0.05). Conclusion:EA at acupoints of Shaoyang meridian can obviously increase the mechanical and thermal pain threshold and improve pain sensitization in CM rats. The mechanism would be related to down-regulating the expression levels of TRPV1 and CGRP mRNA and TRPV1, p-TRPV1 and CGRP protein in the TG of CM rats.
9.Effects of Electroacupuncture at "Fengchi" (GB 20), "Yanglingquan" (GB 34) and "Waiguan" (TE 5) on PPARγ/NF-κB Signalling Pathway in Trigeminal Vascular System in Acute Migraine Model Rats
Zhuozhong HE ; Lei TAN ; Yixiang ZENG ; Shengli ZHOU ; Haojia WEN ; Yang YANG ; Runze TU ; Wei'ai LIU ; Ke HE ; Lei FU
Journal of Traditional Chinese Medicine 2024;65(23):2468-2475
ObjectiveTo investigate the possible mechanism of electroacupuncture at Fengchi (GB 20), Yanglingquan (GB 34) and Waiguan (TE 5) for acute migraine attacks by peroxisome proliferator-activated receptor γ/ nuclear transcription factor-κB pathway in the trigeminal neurovascular system. MethodsForty SD rats were randomly divided into blank group, model group, electroacupuncture group, electroacupuncture plus inhibitor group, and agonist group, 8 rats in each group. Except for the blank group, rats were injected with inflammation decoction intracranial catheter to establish rat models of acute migraine. Thirty minutes after modelling, the electroacupuncture group was given 0.9% NaCl solution by intraperitoneal injection and then electroacupuncture at "Fengchi" (GB 20), "Yanglingquan" (GB 34), and "Waiguan" (TE 5) for 30 mins; the electroacupuncture plus inhibitor group was given PPARγ inhibitor T0070907 (1.5 mg/kg) and eclectroacupuncture as the above for 30 mins; the agonist group was given PPARγ inhibitor pioglitazone hydrochloride (10 mg/kg) for 30 mins. Rats in the blank group and the model group were injected intraperitoneally with 0.9% NaCl solution and then bound and restrained for 30 mins. Behavioural scores were evaluated before modelling, 30 mins after modelling (pre-intervention) and post-intervention; after the last behavioural test, PPARγ, NF-κB p65 mRNA content in rat dura mater was detected by real-time fluorescence quantitative PCR; protein expression of PPARγ, NF-κB p65, interleukin 6 (IL-6), tumour necrosis factor α (TNF-α) in spinal trigeminal nucleus of rats was detected by protein blotting; immunofluorescence double-labelling method was used to detect the fluorescence intensity of PPARγ, NF-κB p65 in spinal trigeminal nucleus of rats. ResultsCompared with the blank group at the same time, the behavioural scores of rats in the remaining groups increased after modelling and after intervention (P<0.01). Compared with the model group at the same time, the beha-vioural scores of rats in the electroacupuncture group, electroacupuncture plus inhibitor group, and agonist group decreased after the intervention (P<0.01). Compared with the electroacupuncture group at the same time, beha-vioural scores reduced in the agonist group and elevated in the electroacupuncture plus inhibitor group after intervention (P<0.01). Compared with the blank group, expression of PPARγ and NF-κB p65 mRNA elevated in the dura mater of rats in the model group, and expression of PPARγ, NF-κB p65, IL-6, and TNF-α protein enhanced in spinal trigeminal nucleus, and the fluorescence intensity of PPARγ and NF-κB p65 enhanced (P<0.01). Compared with the model group, PPARγ mRNA expression in dura mater elevated and NF-κB p65 mRNA expression reduced in each intervention group, PPARγ protein expression in spinal trigeminal nucleus enhanced, and NF-κB p65, IL-6, and TNF-α protein expression weakened; in the electroacupuncture group and the agonist group, PPARγ fluorescence intensity enhanced, and the fluorescence intensity of NF-κB p65 weakened in each intervention group (P<0.05 or P<0.01). Compared with the electroacupuncture group, in the electroacupuncture plus inhibitor group, PPARγ mRNA, protein expression and fluorescence intensity reduced, NF-κB p65 mRNA, protein expression and fluorescence intensity elevated, and IL-6 and TNF-α protein expression enhanced; in the agonist group, PPARγ mRNA and protein expression elevated, NF-κB p65 mRNA and protein expression reduced, and IL-6 and TNF-α protein expression decreased (P<0.05 or P<0.01). ConclusionElectroacupuncture at "Fengchi" (GB 20), "Yanglingquan" (GB 34) and "Waiguan" (TE 5) can can reduce the symptoms of acute migraine attacks in rats, and its mechanism may be related to the up-regulation of PPARγ expression and the reduction of NF-κB expression, thus inhibiting the neurogenic inflammatory response.
10.Advances in crystal nucleation for amorphous drugs
Jie ZHANG ; Kang LI ; Zi-qing YANG ; Zi-han DING ; Sai-jun XIAO ; Zhi-ming YUE ; Li-mei CAI ; Jia-wen LI ; Ding KUANG ; Min-zhuo LIU ; Zhi-hong ZENG
Acta Pharmaceutica Sinica 2024;59(7):1962-1969
Amorphous solid dispersion (ASD) is one of the most effective formulation approaches to enhance the water solubility and oral bioavailability of poorly water-soluble drugs. However, maintenance of physical stability of amorphous drug is one of the main challenges in the development of ASD. Crystallization is a process of nucleation and crystal growth. The nucleation is the key factor that influences the physical stability of the ASD. However, a theoretical framework to describe the way to inhibit the nucleation of amorphous drug is not yet available. We reviewed the methods and theories of nucleation for amorphous drug. Meanwhile, we also summarized the research progress on the mechanism of additives influence on nucleation and environmental factors on nucleation. This review aims to enhance the better understanding mechanism of nucleation of amorphous drug and controlling over the crystal nucleation during the ASD formulation development.

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