1.The relationship between syndrome differentiation of TCM and clinical test results of 196 cases with chronic ;hepatitis B
Chinese Journal of Primary Medicine and Pharmacy 2015;(21):3273-3275,3276
Objective To establish the corresponding relationship between syndrome differentiation of TCM and clinical test results in chronic hepatitis B patients,and thus to provide objective evidence for TCM treatment. Methods 196 patients with chronic hepatitis B,according to the Chinese medicine dialectical typing results were divided into five groups(liver kidney yin deficiency syndrome type 35 cases,Shire Zhongzu syndrome type 24 cases, Yang deficiency of spleen and kidney syndrome type 26 cases,damp heat resistance type and blood stasis syndrome type 42 cases,liver stagnation and spleen deficiency syndrome type 69 cases).HBV -DNA,hepatitis B,TBIL,ALT, TTT were detected.Results In the HBV -DNA test,the majority of the five groups were positive,the overall compar-ison,the difference was not statistically significant (P =0.937).But in the second liver five,TBIL,ALT and TTT detection,the detection results were reflected in the corresponding relationship between TCMsyndrome differentiation. Such as in TTT detection,liver kidney yin deficiency syndrome type and liver stagnation and spleen deficiency syn-drome type group of patients,most patients with TTT detection results between 7 and 10,two groups sequentially com-pared with the other three groups,the differences were statistically significant (all P =0.000);damp heat resistance type and blood stasis type in the patients group,TTT detection results were generally less than 6,two groups in turn compared with the other three groups,the differences were statistically significant (all P =0.000);Yang deficiency of spleen and kidney in the patients group,TTT detection results were generally more than 10,and compared with the other four groups,the differences were statistically significant (all P =0.000).Conclusion TCMsyndrome differen-tiation of chronic hepatitis B patients has a certain correlation with the clinical test results.
2.Effect of Dendrobium officinale Polysaccharides in Reversing Hepatic Fibrosis in Rats via Notch Signaling Pathway and Ultrasound Evaluation
Ying XU ; Jina LUO ; Liuqing YANG ; Qianqian TIAN ; Jiamao CHENG ; Haiyan CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(21):70-77
ObjectiveTo investigate the effect of Dendrobium officinale polysaccharides (DOP) in reversing carbon tetrachloride (CCl4)-induced hepatic fibrosis (HF) in rats via the Notch signaling pathway and evaluate the therapeutic effect of DOP by ultrasound elastography. MethodFifty-six male SD rats were randomized into normal, model, colchicine (1×10-4 g·kg-1), Fuzheng Huayu powder (0.45 g·kg-1), and low-, medium-, and high-dose (0.05, 0.1, 0.2 g·kg-1) DOP groups (n=8). The rats in the model group and each treatment group were injected subcutaneously with a mixture of CCl4-olive oil (2∶3) once every 3 days for 10 weeks. After 6 weeks of modelling, the rats were administrated with corresponding drugs once a day for 4 weeks. Hematoxylin-eosin (HE) staining and Masson staining were employed to observe the pathomorphological changes of the liver tissue. An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bile acids (TBA). Enzyme-linked immunosorbent assay (ELISA) was employed to measure the serum levels of hyaluronic acid (HA), laminin (LN), type Ⅲ precollagen (PC-Ⅲ), and type Ⅳ collagen (Col-Ⅳ). The mRNA and protein levels of α-smooth muscle actin (α-SMA), Notch1, Jagged1, and Hes1 in the liver tissue were determined by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. The Young's modulus (YM) of the rat liver was measured by acoustic radiation force impulse (ARFI) elastography before and after treatment. Then, the correlations of YM with the serum levels of HA, LN, PC-Ⅲ, and Col-Ⅳ and the protein levels of α-SMA and Notch1 signaling pathway-related factors in the liver tissue were analyzed. ResultCompared with the normal group, the model group showed disordered arrangement of liver cell cords, obvious infiltration of inflammatory cells, appearance of a large number of fat vacuoles, and fibrous proliferation, elevated levels of ALT, AST, TBA, ALP, HA, LN, PC-Ⅲ, and Col-Ⅳ in the serum, and up-regulated mRNA and protein levels of α-SMA, Notch1, Jagged1, and Hes1 in the liver tissue (P<0.01). Compared with the model group, drug interventions alleviated the pseudolobule formation and the collagen deposition in confluent areas. Except that the serum level of ALT in the low-dose DOP group had no significant changes, drug interventions, especially high-dose DOP, lowered the levels of ALT, AST, TBA, ALP, HA, LN, PC-Ⅲ, and Col-Ⅳ in the serum and down-regulated the mRNA and protein levels of α-SMA, Notch1, Jagged1, and Hes1 in the liver tissue (P<0.05, P<0.01). The results of ARFI and correlation analysis showed that the YM of the liver tissue was increased in the model group (P<0.01) compared with that in the normal group, Compared with the model group, drug interventions decreased YM (P<0.01). YM was positively correlated with the expression levels of HA, LN, PC-Ⅲ, α-SMA, Notch1, Jagged1, and Hes1s (r=0.754, 0.734, 0.801, 0.885, 0.896, 0.757, and 0.800, respectively, P<0.01), and it had a moderate correlation with Col-Ⅳ (r=0.688, P<0.01). ConclusionDOP can reverse HF by down-regulating the Notch1/Jagged1/Hes1 signaling pathway. YM can be used as an indicator in the assessment of the efficacy of DOP against HF.
3.Dendrobium Officinale Polysaccharide Alleviates Hepatic Fibrosis via Epithelial-Mesenchymal Transition
Liuqing YANG ; Qin FAN ; Yajie BAI ; Ying XU ; Jina LUO ; Jiamao CHENG ; Haiyan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(1):76-84
ObjectiveTo investigate the effect of Dendrobium officinale polysaccharide (DOP)on CCl4-induced hepatic fibrosis(HF)and its mechanism. MethodsA total of 56 male SD rats were randomly divided into seven groups: normal group(NG),model group(MG),colchicine group(CG, 0.1 mg/kg), Fuzheng Huayu group(FG, 0.45 g/kg),low-dose DOP group(LDG, 0.05 g/kg),middle-dose DOP group(MDG, 0.1 g/kg)and high-dose DOP group(HDG,0.2 g/kg),with 8 rats in each group. HF rat model was established by subcutaneous injection with 40% CCl4 olive oil mixture, every 3-day for 10 weeks. At the end of the sixth week, the drug groups were treated with colchicine, Fuzheng Huayu and DOP solution by gavage respectively, once a day for 4 weeks. NG and MG groups were similarly handled with an equal amount of 0.9 % normal saline. Liver histopathology was detected using hematoxylin-eosin (HE), Masson and Sirius red staining; blood biochemistry was tested for liver function and four indicators of HF; RT-qPCR and Western Blot were used to measure the expression of α-SMA, Col-I, E-cadherin, and ZEB1 genes and proteins in the liver tissues of rats, respectively. ResultsHE, Masson, and Sirius red staining showed that the liver tissue of MG rats had typical pathologic features of HF, and the degree of HF was alleviated in LDG, MDG, and HDG rats, respectively. Liver function test results showed that the serum AST, TBIL, and AKP levels were significantly lower in LDG, MDG, and HDG, compared with those of the MG (P < 0.05 or < 0.01). Meanwhile, ALT levels in serum deceased remarkably except in LDG (P < 0.05 or < 0.01). The four results of HF showed that the serum HA, LN, PC-Ⅲ, and COL-Ⅳ levels in LDG, MDG, and HDG rats were significantly decreased compared with those of the MG (P < 0.05 or < 0.01). The relative expressions of α-SMA, COL-I, and ZEB1 genes and proteins were significantly decreased in the liver tissues of LDG, MDG, and HDG (P < 0.05 or < 0.01), and the relative expression of E-cadherin gene and protein increased (P < 0.05 or < 0.01). In addition, the expressions of HA, α-SMA, COL-I, ZEB1 and E-cadherin were dependent on the dose of DOP. ConclusionDOP alleviated the degree of CCl4 induced HF in rats by inhibiting the epithelial-mesenchymal transition in liver tissue.
4.Therapeutic Effect of Canagliflozin on Nephrotic Syndrome and Its Ultrasonic Evaluation
Wen-juan HONG ; Hong-jun LI ; Jiu-lin ZOU ; Wei HUAN ; Xiao LI ; Jia-mao CHENG ; Hai-yan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2023;44(1):71-77
ObjectiveTo investigate the therapeutic effect of antidiabetic drug canagliflozin (CGLZ) on adriamycin-induced nephrotic syndrome (NS) in rats, and the evaluation of contrast-enhanced ultrasound (CEUS) combined with color Doppler flow imaging (CDFI) during the treatment. MethodsA total of 56 male SD rats were randomly divided into normal group (NG), model group (MG), prednisone (PAT) group (PG), low-dose single CGLZ group (LSCG), high-dose single CGLZ group (HSCG), low-dose CGLZ + PAT group (LUCG) and high-dose CGLZ + PAT group (HUCG), with 8 rats in each group. The NS model in rats was induced by injecting adriamycin twice into the tail vein, and then the NS rats were treated by intragastric administration daily for 6 weeks with reference of PAT. Twenty-four hour urine total protein (24 h-UTP) was assessed one day before the start of oral administration and at the end of 2, 4 and 6 weeks after oral administration, respectively. CDFI and CEUS were performed on the right renal artery at the end of 6 weeks after oral administration, and the blood of abdominal aorta was taken for serological test the next day. ResultsCompared with those detection index of NG rats, the 24-hour UTP of MG rats increased (P<0.01), the serum ALB decreased and TG, TC, LDL increased (P<0.01), and CDFI shows that RRCT was thinner (P<0.01) and the renal artery blood flow indicators RA-PI, RA-RI, RA-S/D all increased (P<0.05), and CEUS image shows that the TIC curve parameters TTP, AT, AUC all increased and DPI decrease in MG rats (P<0.01). After drug treatment, compared with those detection index of MG rats, 24 h-UTP decrease in LSCG after 2 weeks (P<0.01), and decrease significantly in all drug groups after 6 weeks (P<0.01); the serological test results show that the serum ALB in all CGLZ groups increased (P<0.05), TG decrease in LSCG (P<0.01), TC and LDL also decrease in LUCG after 6 weeks (P<0.05); CDFI shows that the RRCT thinning degree in all CGLZ is reduced (P<0.01), and the RA-PI in LSCG, RA-RI in PG, and RA-S/D in PG, LSCG, HSCG and LUCG rats all decreased (P<0.05); CEUS shows that the TTP, AT and AUC of renal TIC curve in drug treatment groups all decreased (P<0.01), and the DPI in PG, HSCG, LUCG and HUCG rats increased (P<0.01). ConclusionsCGLZ has the effect of treating NS, and the small dose is the best. CEUS combined with CDFI can be used to evaluate the renal morphology and hemodynamic changes of NS model rats before and after drug treatment, which is helpful to guide clinical application.
5.Antioxidant and Anti-tumor Effect and Mechanisms of Andrographis paniculata Polysaccharide on Human Cervical Cancer HeLa Cells
Lijin HUANG ; Zilin LI ; Ziyan YANG ; Han WANG ; Guiyuan CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):80-88
ObjectiveTo explore the antioxidant and anti-human cervical cancer HeLa cell effect and mechanisms of Andrographis paniculata polysaccharide (APP). MethodCell function assays were conducted to assess the effects of APP (400, 450, 500 mg·L-1) on the proliferation, migration, and invasion of HeLa cells using the cell counting kit-8 (CCK-8) assay, scratch assay, and Transwell assay. Molecular mechanism experiments were conducted to detect the effects of APP on HeLa cell apoptosis and cell cycle-related mRNA and protein expression using flow cytometry, real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR), and Western blot analysis. The antioxidant activity of APP was tested using DPPH+, OH-, and reducing power assays. ResultCompared with the blank group, APP (400, 450, 500 mg·L-1) significantly inhibited the migration, proliferation, and invasion abilities of HeLa cells in a time- and dose-dependent manner (P<0.05, P<0.01). Flow cytometry with propidium iodide (PI) single staining was used to detect cell cycle. The results showed that compared with the blank group, the proportion of HeLa cells in the G2/M phase increased after 48 hours of treatment with APP (400, 450, 500 mg·L-1), indicating that APP can arrest HeLa cells in the G2/M phase. Flow cytometry with fluorescein isothiocyanate (Annexin V-FITC)/PI apoptosis kit was used to detect cell apoptosis. Compared with the blank group, the proportion of early and late apoptotic HeLa cells increased in a dose-dependent manner after 48 hours of APP (400, 450, 500 mg·L-1) treatment (P<0.05, P<0.01), indicating that APP promotes HeLa cell apoptosis. The results of Real-time PCR and Western blot assay showed that compared with the blank group, after 48 hours of APP (400, 450, 500 mg·L-1) treatment resulted in decreased mRNA and protein expression of cell cycle-dependent kinase-1 (CDK-1), Cyclin B1, and B-cell lymphoma-2 (Bcl-2), and increased mRNA and protein expression of cysteine aspartate protease (Caspase)-3, Caspase-8, Caspase-9, and Bcl-2-associated X protein (Bax) (P<0.05, P<0.01). These findings were consistent with the flow cytometry results and showed a dose-dependent effect. In vitro antioxidant tests demonstrated that different concentrations of APP (50-1 000 mg·L-1) were able to scavenge DPPH+ and OH- radicals, indicating certain antioxidant activity. ConclusionAPP possesses antioxidant activity and can inhibit the viability of HeLa cells while promoting their apoptosis.
6.Anti-tumor Effect and Mechanism of Lycopus lucidus Polysaccharide on Non-small Cell Lung Cancer A549 Cells
Ting-ting CHENG ; Yan LI ; Gui-yuan CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):83-90
ObjectiveTo study the anti-tumor activity and mechanism of Lycopus lucidus polysaccharide (LLP) in vitro. MethodCell counting kit-8 (CCK-8) assay was used to detect the inhibitory effect of LLP (0, 5, 10, 15, 20 g·L-1) on the proliferation of A549 cells at different time points (24,48,72 h). The migration and invasion abilities of A549 cells were detected by wound healing assay and transwell assay after LLP (10, 20 g·L-1) treatment for 24,48 h. Propidium iodide (PI) single staining was applied to determine the effect of LLP of different concentrations (10,20 g·L-1) on the cell cycle of A549. The apoptosis of A549 cells induced by LLP (10, 20 g·L-1) was detected by Annexin V-FITC/PI kit. Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) was adopted to measure effect of LLP (10, 20 g·L-1) on gene expression of cysteine aspartate protease-3 (Caspase-3),cysteine aspartate protease-8 (Caspase-8),cysteine aspartate protease-9 (Caspase-9),cyclin-dependent kinase-1 (CDK-1), and Cyclin B1 in A549 cells. Western blot was used to detect the effect of LLP on protein expression of Caspase-3,Caspase-8,Caspase-9,B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax),CDK-1,cyclin-dependent kinase-4 (CDK-4),cyclin-dependent kinase-6 (CDK-6),Cyclin B1,and Cyclin D1 in A549 cells. ResultCompared with the blank group, the LLP group showed decreased proliferation, migration, and invasion of A549 cells (P<0.05, P<0.01), increased proportion of G0/G1 phase (P<0.05), enhanced apoptosis rate (P<0.05, P<0.01), elevated mRNA expression of Caspase-3,Caspase-8,and Caspase-9 (P<0.05,P<0.01), reduced mRNA expression of CDK-1 and Cyclin B1 (P<0.05,P<0.01), up-regulated protein expression of Caspase-3,Caspase-8,Caspase-9, and Bax (P<0.05, P<0.01), and down-regulated protein expression of Bcl-2, CDK-1, CDK-4, CDK-6, Cyclin B1, and Cyclin D1 (P<0.05, P<0.01). ConclusionLLP can inhibit the proliferation of A549 cells, block the cell cycle in the G0/G1 phase (also G2/M phase), and induce cell apoptosis via the mitochondrial apoptosis pathway and death receptor pathway.
7.Two-staged immediate procedure of breast reconstruction with tissue expansion
Dali MU ; Jie LUAN ; Baoning ZHANG ; Ge CHEN ; Dou JIN
Chinese Journal of Medical Aesthetics and Cosmetology 2012;(6):416-419
Objective To present our experience in two-stage procedure immediate breast reconstruction,which aimed to retain local tissue condition for breast reconstruction,overcome the limitation of radiotherapy on implant reconstruction,and simplify the procedure as well as improve cosmetic result.Methods The proceduces of our method were divided into two stages:in the first stage,the round-shaped expander was implanted in the subpectoral major space during the procedure of mastectomy; at the same time as expander implanted,the first saline injection was performed; with 4 to 8 weeks of inflation,the tissue expander offered adequate tissue for breast reconstruction; in the second stage,silicon gel implant,latissimus dorsi muscle flap,extended latissimus dorsi muscle flap and deep inferior epigastic perforatior flap were used for breast reconstruction after the tissue expander was exchanged.Results Thirty-four patients had undergone the two-staged breast reconstruction using tissue expansion.The average time of therapy was 5.5 months.There were no postoperative complications such as implant exposure,additional scar,and flap necrosis.The follow-up time was 6 to 18 months and the result showed excellent contour of the breast at a satisfactory rate of 97.1%.Conclusions Two-staged technique using tissue expansion in breast reconstruction is easily done and the complications are rare.The technique possesses advantages such as avoiding affection of radiotherapy on silicongel implant and elasticity of skin,retaining maximal local tissue for breast reconstruction.There is no necessary for correction of skin defect and additional scarring,thus the patch-like appearance of breast is avoided.The patients need not to experience depression post mastectomy.Two-staged procedure immediate breast reconstruction is a safe and reliable technique that is especially applicable to the patients who need radiotherapy.
8. Consistency evaluation method for Lianhua Qingwen Capsule based on microcalorimetry
Chinese Traditional and Herbal Drugs 2017;48(11):2202-2206
Objective: To study the effect of Lianhua Qingwen Capsule (LQC) on the growth and metabolism of Pseudomonas aeruginosa and to establish a new method to evaluate the consistency of LQC. Methods: The power-time curves and corresponding thermodynamic parameters such as appearance time (T), total heat output (Q), maximum power output (P), and growth rate constant (k) of P. aeruginosa were determined by microcalorimetry. The regression analysis was carried out with T, P, Q, and k as parameters, and the optimal parameters were selected according to the correlation coefficient to calculate the antibacterial effective rate. The effects of 11 batches of LQC and specially treated LQC samples on growth and metabolism of P. aeruginosa were determined by an index antibacterial effective rate to evaluate the antibacterial activity of LQC. Results: LQC had obvious inhibitory effect on P. aeruginosa in the range of 10.00 to 47.68 mg/mL, and the bacteriostatic efficacy and dosage of LQC had good correlation. By regression analysis, it was found that T was the best parameter in the four thermodynamic parameters, r = 0.990. The change of this parameter could directly evaluate the effect of LQC on the growth and metabolism of P. aeruginosa, with good stability. The appearance time was selected as a parameter to calculate its antibacterial effective rate (E). The results showed that there was no significant difference in E of different batches of LQC samples. And the E value was significantly changed when the sample was placed in a humid/high temperature environment. Conclusion: The microcalorimetric method can be used to evaluate the antibacterial activity and the quality consistency of LQC capsule, which has the high precision and good accuracy. The study has provided a new method to evaluate the quality consistency of TCM.
9. Chemical Constituents of the Roots of Astragalus acaulis
Chinese Pharmaceutical Journal 2020;55(15):1253-1258
OBJECTIVE: To study the major chemical constituents of the roots of Astragalus acaulis. METHODS: Compounds were isolated and purified by using silica gel, Sephadex LH-20, and Rp-18 chromatographic techniques. Their structures were determined based on various spectral analysis and physicochemical properties. RESULTS: Ninteen compounds were obtained from ethyl acetate and n-butanol fractions derivated from the methanol extract of the roots of A. acaulis, and identified as liquiritigenin (1), isoliquiritigenin (2), isomucronulatol (3), syringaresinol-4-O-β-D-glucopyranoside (4), lariciresinol (5), lariciresinol-4-O-β-D - glucopyranoside (6), 5-methoxylariciresinol-4-O-β-D-glucopyranoside (7), p-hydroxybenzaldehyde (8), sorbitol (9), 7-oxositosterol (10), 7β-hydroxysitosterol (11), β-sitosteryl-D-glucoside-6'palmitate (12), 5α, 8α-epidioxy-ergosta-6, 22E-dien-3β-ol (13), 2α, 3β-dihydroxyloleanolic acid (14), methyl stearate (15), β-sitosterol (16), daucosterol (17), oleic acid (18), and methyl elaidate (19). CONCLUSION: All of the compounds reported in this study are isolated from the plant for the first time. A. acaulis is rich in flavonoids, lignans, and steroids.
10.Selection of breast implant during immediate breast reconstruction after mastectomy
Ge CHEN ; Chunwei XIE ; Dali MU ; Jie LUAN
Chinese Journal of Clinical Oncology 2014;(16):1049-1051
Objective:To demonstrate the selection of breast implant during an immediate breast reconstruction post-mastectomy and analyze the indication of this technique. Methods:From June 2007 to June 2012, a total of 121 patients with breast cancer received immediate breast reconstruction with breast implants. Among the 121 patients, 89 patients had simple mastectomy, while the rest under-went modified radical mastectomy in the Department of Breast Neoplasm, Nanchang No.3 Hospital. The volumes of the resected breast tissues were measured using Archimedes principle. The diameters of the tissues were also determined. Proper breast implants were se-lected according to the measured data. Results: Postoperative complications, such as implant exposure, flap necrosis, and infection, were not found. Follow-up period ranged from 6 months to 12 months. Patients answered a questionnaire that displayed their degree of satisfaction for the breast operation outcome. Results show that 89.3%of the patients (108/121) were very satisfied, 9%were (11/121) satisfied, and 1.7%(2/121) were unsatisfied. Conclusion:Immediate breast reconstruction with breast implant post-mastectomy is an ideal method for rebuilding the breast. This technique is advantageous because it prevents damage to the donor site and retains the maxi-mal elasticity of the skin for breast reconstruction. Accurate parameters of breast implants, which are important to achieve good surgical results, could be obtained using Archimedes principle.