1.The mechanisms of paeonol reversing multidrug resistance in ovarian cancer SKOV3/DDP cells
Li HAN ; Xiao-juan GUO ; Zhong CHEN ; Hua BIAN ; Chao-yun ZHANG ; Wen-hua ZANG ; Qian WANG ; Jiu-lue HU
Acta Pharmaceutica Sinica 2018;53(9):1511-1517
The purpose of this research is to investigate the effects and mechanisms of paeonol (PL), a phenolic compound found in many traditional Chinese formulations, on reversing drug resistance in the ovarian cancer resistant SKOV3/DDP cells. The results showed that PL had significant drug-resistant reversal effect on SKOV3/DDP cells. Flow cytometry showed that PL could inhibit P-glycoprotein (P-gp) function in a concentration-dependent manner. Fluorescent quantitative PCR and cell immunofluorescence techniques were used to detect mechanisms of action. Results revealed that both the inhibitory effect on MDR1/P-gp and metadherin (MTDH) expression and the induction effect on phosphatase and tensin homolog (PTEN), by 15, 30, and 60 μmol·L-1 PL, were increased with increased concentrations of PL (P < 0.01, P < 0.05). The inhibitory effect on MTDH mRNA and the induction effect on PTEN mRNA, by PI3K inhibitor LY294002, were stronger or equivalent to that of the 60 μmol·L-1 PL treated group; however, the inhibition or induction effect on MTDH or PTEN protein were only comparable to the 15 μmol·L-1 PL treated group. The present study shows that the effect of PL on SKOV3/DDP cells may be related to the inhibition of P-gp function and expression, the inhibition of MDR1, MTDH expression, and the induction of PTEN expression, all which can provide a theoretical foundation for PL as a drug resistance reversal agent on the treatment of ovarian cancer chemotherapy resistance.
2.Study on the mechanism of Wenyang huazhuo tongluo formula contained serum inhibiting the proliferation of Th17 cells in scleroderma patients
Kelei GUO ; Yingli LI ; Bo BIAN ; Li HAN ; Kai LI ; Hong ZHANG ; Hua BIAN
China Pharmacy 2022;33(15):1820-1824
OBJECTIVE To study the effects of Wenyang huazhuo tongluo formula conta ined serum on the expression and methylation of retinoic acid related nuclear orphan receptor (RORγt),and to explore the mechanism of its regulation of Th 17 cell proliferation in the treatment of scleroderma. METHODS Wistar rats were given 15,30,60 g/(kg·d)Wenyang huazhuo tongluo formula intragastrically to prepare different doses of drug-contained serum ,and peripheral blood of scleroderma patients were collected to sort Th 17 cells. Using blank serum as blank control ,methyltransferase inhibitor decitabine (DCA)as positive control , Th17 cells were treated with different concentrations of drug-contained serum ,and then the proliferation of Th 17 cells was detected by CCK- 8;mRNA expressions of RORγt and IL-17 were detected by qRT-PCR ,and the protein expression of RORγt was detected by Western blot assay ;the protein expression of IL- 17 in the cell supernatant was detected by ELISA ,and the methylation level of RORγt gene promoter was detected by methylation-specific PCR. The transcriptional activity of RORγt gene promoter was detected by dual-luciferase assay. RESULTS Compared with blank control ,Wenyang huazhuo tongluo formula contained serum and DCA could inhibit the proliferation of Th 17 cells(P<0.05),reduced the mRNA and protein expressions of RORγt and IL-17,enhanced the methylation level of RORγt gene promoter and attenuated , the transcription activity of RORγt gene promoter. Except for mRNA expression of Th 17 and the methylation level of RORγt gene promoter in drug-contained serum low-dose group ,there were statistical significance in above indexes of other groups(P<0.05). CONCLUSIONS Wenyang huazhuo tongluo formula can promote the methylation lev el of RORγt gene,and inhibit the expression of RORγt and the secretion of IL-17,so as to inhibit the proliferation of Th 17 cells.
3. Six body indexes and typing characteristics of Xibe adults
Guo-Chang XU ; Zhi-Guo CHEN ; Guo-Chang XU ; Zhi-Guo CHEN ; Ke-Lei GUO ; You-Feng WEN ; Dong JIANG
Acta Anatomica Sinica 2023;54(4):477-483
[Abstract] Objective To explore the body index and typing characteristics of Xibe nationality. Methods According to Anthropomatric Methods, height, chest circumference, sitting height, shoulder width, pelvic width and body mass morphological indexes were collected from 588 Xibe people in Yili area and 420 Xibe people over 18 years old in Shenyang area. After data collation, body index and typing were calculated and analyzed statistically. Results Except for Xibe male stature-shoulder breadth index and Caup’ s index in Yili area and female stature-shoulder breadth index in Shenyang area, there were differences among age groups in other indexes (P < 0. 05). Except for male stature-chest circumference index, stature-crista iliaca index, female stature-chest circumference index, stature-sitting height index, Manouvrier’s skelic index, there were differences among regions (P<0. 05). The adult body shape of Xibe nationality was mainly of wide chest, wide shoulder, long trunk, wide pelvis, medium leg and sub-long leg (male), sub-short leg and medium leg (female), and overweight. Conclusion There are regional and age differences in the body index and typing of Xibe adults. Compared with other ethnic minorities, Xibe nationality’s stature-chest circumference index, stature-shoulder breadth index, stature-crista iliaca index, Caup’s index are at a higher level. The proportion of legs and body of Xibe nationality is in the middle level, while the proportion of trunk is slightly shorter for men and slightly longer for women.
4.Optimization of extraction process and antioxidant activity of total phenolic acids from Cibotii rhizoma
Zhongjing GUO ; Xinjun XU ; Zhimin ZHAO ; Depo YANG
China Pharmacist 2024;27(1):8-16
Objective To optimize the extraction process of total phenolic acids from Cibotii rhizoma using Box-Behnken design-response surface methodology and evaluate antioxidant activity of total phenolic acid in vitro.Methods Taking the liquid-solid ratio,ethanol concentration and extraction temperature as influencing factors,and the extraction rate of total phenolic acids as evaluation indicator,the extraction process of total phenolic acids from Cibotii rhizoma was optimized using a three-factor and three-level Box-Behnken design-response surface methodology on the basis of single factor tests.Meanwhile,the scavenging effects of total phenolic acid extract from Cibotii rhizoma on ABTS·+and DPPH· was determined.Results The optimized extraction process for total phenolic acids from Cibotii rhizoma was as follows:the ethanol concentration of 55%,the extraction temperature of 88℃ and the liquid-solid ratio of 60∶1 mL/g.Under these conditions,the extraction rate could reach 8.67%.When the mass concentration of total phenolic acids extract were 2 mg/mL and 1 mg/mL,the clearance rates of ABTS·+and DPPH·were 92.76%and 88.66%,respectively.Conclusion The theoretical values obtained from the response surface optimization method are consistent with the actual measured values,and the extraction process of total phenolic acids from Cibotii rhizoma was simple and feasible.The total phenolic acids extract from Cibotii rhizoma exhibit strong antioxidant activity in vitro.
5.Guizhi Fuling Wan reverses drug resistance by regulating PTEN and MTDH in ovarian cancer SKOV3/DDP Cells
Xiao-Juan GUO ; Li HAN ; Wen-Hua ZANG ; Qian WANG ; Hua BIAN ; Jiu-Lue HU ; Chao-Yun ZHANG
Chinese Journal of Pharmacology and Toxicology 2018;32(4):284-284
OBJECTIVE To detect the reversal effect of Guizhi Fuling Wan on cisplatin-resistant ovarian cancer SKOV3/DDP cells and its relationship with protein expression of phosphatase and tensin homolog (PTEN) and metadherin (MTDH). METHODS Guizhi Fuling Wan (GFW) concentrated solution was prepared according to the Chinese Pharmacopoeia 2015 edition, Wistar rats were given GFW viagavage at 4 g·kg-1·d-1,8 g·kg-1·d-1,16 g·kg-1·d-1,or given saline as blank control for 5 days.Blood samples were taken and the corresponding drug-containing low-dose sera, medium-dose sear, high-dose sera and blank sera were prepared.The XCELLigence RTCA S16 real-time label-free cell analyzer was used to detect the reversal effect by the sera combined with cisplatin or paclitaxel in SKOV3/DDP cells. Annexin V-FITC and PI double-staining were used to detect the apoptosis-inducing effect of the sera in the cells. RT-qPCR and western blot were used to detect the mRNA and protein expression of PTEN and MTDH after the cells treated with the sera. RESULTS The inhibition rate of low-dose sera against SKOV3/DDP cells was less than 5%.After the low-dose sera combined with cisplatin or pacli-taxel, the IC50 of SKOV3/DDP cells against cisplatin and paclitaxel decreased by 3.01 and 1.79-fold, respectively.The total apoptosis rates induced by the low-dose sera,medium-dose sear,high-dose sera and blank sera in SKOV3/DDP cells were 11.08±0.13,19.42±0.30,24.23±0.31,and 3.21±0.24,respec-tively; there was a significant difference between the groups (P<0.01). RT-qPCR results showed that, compared with the blank serum, the sera can up-regulate the expression of PTEN mRNA and down-regulate the expression of MTDH mRNA in a dose-dependent manner. Western blot results showed that the induction effect to PTEN protein and the inhibition effect to MTDH protein by the sera were gradually enhanced with thesera dose increasement. CONCLUSION The resistance reversal effect of Guizhi Fuling Wan on ovarian cancer SKOV3/DDP cells may be related to the inhibition of MTDH, up-regulation of PTEN and induction of apoptosis, providing with an experiment basis for the applica-tion of Guizhi Fuling Wan as a reversal agent for chemotherapy resistance of ovarian cancer.
6.Research Progress on Chemical Constituents,Pharmacological Effects and Quality Control of Cibotii Rhizoma
Zhongjing GUO ; Ennian LI ; Zhimin ZHAO ; Depo YANG ; Xinjun XU
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(1):152-157
Cibotii Rhizoma has a long history of folk usage and clinical applications,which is commonly used to treat symptoms such as waist and knee soreness,numbness of hands and feet.Its chemical components mainly include sugars and glycosides,aromatics,volatile oils,pterosins,flavonoids,steroids,pyranones,amino acids and inorganic elements.Pharmacological studies have shown that Cibotii Rhizoma has antioxidant,anti-inflammatory,anti-osteoporosis,gastrointestinal protection,antibacterial,liver protection,anticancer,analgesic and other effects.This paper systematically reviews the research status of chemical constituents,pharmacological effects and quality control of Cibotii Rhizoma,which provides reference for the development and rational use of Cibotii Rhizoma.
7.Wenxiao Powder Alleviates Depression by Promoting Neurogenesis via BDNF/TrkB/ERK/CREB Signaling Pathway
Duo ZHANG ; Xiuhui GUO ; Yucheng LI ; Yunli GAO ; Ming BAI ; Xiangli YAN ; Erping XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):100-108
ObjectiveTo decipher the mechanism of Wenxiao powder in alleviating corticosterone-induced depression-like behaviors in mice. MethodMale ICR mice were randomized into normal, model, paroxetine (20 mg·kg-1), and low- and high-dose (3.27, 6.54 g·kg-1, respectively) Wenxiao powder groups. The mice in normal and model groups received equal volume of saline. Other groups except the normal group were injected with corticosterone subcutaneously 0.5 h after gavage to induce depression. Mice were tested for depression-like behaviors after drug administration. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the corticosterone content in the serum. Nissl staining was performed to observe the damage of hippocampal neurons. Immunofluorescence staining was employed to observe the expression of double cortin (DCX) in the dentate gyrus (DG) of the hippocampus. Western blot was employed to determine the expression of proteins in the brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB)/extracellular signal-regulated kinase (ERK)/cAMP-response element-binding protein (CREB) pathway in the hippocampus. ResultCompared with the normal group, the model group showed decreased sucrose preference rate, increased immobility time in the tail suspension test (P<0.01), and reduced residence time in the central area of the open field and the total movement distance (P<0.05, P<0.01). In addition, the modeling elevated the corticosterone level in the serum (P<0.01), decreased the volume and intensified the nuclear staining of hippocampal neurons in the DG area, reduced the expression of DCX in the DG area, and down-regulated the protein levels of BDNF, phosphorylated (p)-TrkB, p-ERK, and p-CREB in the hippocampus (P<0.05, P<0.01). Compared with the model group, low-dose Wenxiao powder improved the mouse behavivors in the sucrose preference, open field, and tail suspension tests (P<0.05, P<0.01), and high-dose Wenxiao powder improved the behaviors in the sucrose preference and open field tests (P<0.05, P<0.01). In addition, Wenxiao powder lowered the serum corticosterone level (P<0.01) and recovered the structure and morphology of neurons with obvious nuclei and presence of Nissl bodies in the DG area of the hippocampus. Moreover, Wenxiao powder at both doses promoted the expression of DCX in the DG area, and high-dose Wenxiao powder up-regulated the protein levels of BDNF, p-TrkB, p-ERK, and p-CREB in the hippocampus (P<0.05, P<0.01). ConclusionWenxiao powder can alleviate corticosterone-induced depression-like behaviors and promote neurogenesis in mice possibly by activating the BDNF/TrkB/ERK/CREB signaling pathway.
8.Improvement on Quality Standard of Belladonnae Herba
Yong LIU ; Ya-lin LIU ; Xue JIANG ; Jin-miao WANG ; Qing-mei FENG ; Hui-min GAO ; Zhen-guo LI ; Er-ping XU ; Li-ping DAI ; Zhi-min WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(2):175-181
ObjectiveTo improve the current standard of Belladonnae Herba in the 2020 edition of Chinese Pharmacopoeia. MethodTaking hyoscyamine sulfate, atropine sulfate and scopoletin as reference substances, and ethyl acetate-methanol-concentrated ammonia(17∶4∶2)as developing solvent, thin layer chromatography (TLC) was applied in the qualitative identification of Belladonnae Herba. The moisture, total ash and ethanol-soluble extract of Belladonnae Herba were determined based on the general principles in the 2020 edition of Chinese Pharmacopoeia (volume Ⅳ). The contents of hyoscyamine sulfate and scopolamine hydrobromide were analyzed by high performance liquid chromatography (HPLC) with mobile phase of acetonitrile-54 mmol·L-1 phosphate buffer solution (14∶86), flow rate of 1.0 mL·min-1 and detection wavelength at 210 nm. ResultThe spots in the TLC were clear with good separation and specificity. Hyoscyamine sulfate and scopolamine hydrobromide showed a good linearity with peak area in the range of 0.024 7-0.789 6 g·L-1 (r=0.999 9) and 0.003 9-0.124 0 g·L-1 (r=0.999 9), the average recoveries of these two ingredients were 100.29% (RSD 1.6%) and 99.04% (RSD 1.4%), respectively. The limits for moisture, total ash in Belladonnae Herba should be less than 13.0% and the limit for the ethanol-soluble extract should be more than 10.0%. Due to the low content and wide variation of scopolamine hydrobromide, the content of hyoscyamine sulfate should not be less than 0.098%. ConclusionThe established method is simple, specific and reproducible, which can be used to improve the quality control standard of Belladonnae Herba.
9.Clinical effect of dual growth rod technique in the treatment of early-onset scoliosis
Zhongjing JIANG ; Minzhi LIU ; Ang DENG ; Chaofeng GUO ; Hongqi ZHANG
Chinese Journal of Orthopaedics 2022;42(18):1220-1229
Objective:To investigate the clinical efficacy of dual growing rods (DGR) in the treatment of early onset scoliosis (EOS), and to evaluate the safety of its clinical application.Methods:From March 2015 to August 2021, a total of 20 EOS patients with onset age within 10 years old who were treated with dual growth rod technique were retrospectively analyzed, including 8 males and 12 females. The mean age of patients at first surgery was 9.0±1.4 years (range, 6.3-10.8 years); the preoperative Cobb angle was 59.0°±16.8° (range, 41.2°-103°). The Cobb angle, thoracic kyphosis angle, lumbar lordosis angle, T 1-T 12 height, T 1-S 1 height, coronal plane imbalance, sagittal vertical axis (SVA), apical vertebral translation (AVT) and Campbell's space available for lung ratio (SAL) were recorded and analyzed while surgery-related complications were also recorded. Results:The average follow-up time of 20 patients was 28.07±14.30 months. The average initial hospital stay was 20.56±8.28 days, the average initial operation time was 211.70±39.80 min, the average blood loss in the initial operation was 255.00±149.50 ml, and the average surgical distraction was 1.60±0.51 times per person. The Cobb angle of the main curve in the coronal plane decreased from 59.00°±16.80° before operation to 33.40°±11.80° after the initial operation, which was 29.67°±11.67° at 1 year and 29.40°±11.30° at the last follow-up. Preoperative thoracic kyphosis angle was 41.39°±6.06°, decreased to 31.72°±3.56° after the initial operation, was 30.32°±4.26° at 1-year and 30.24°±4.23° at the last follow-up; preoperative lumbar lordosis angle was 45.90°±8.03°, decreased to 42.65°±9.05° after initial operation, 41.55°±7.84° at 1-year follow-up, and 41.53°±8.21° at the last follow-up; preoperative T 1-S 1 height was 31.76±4.42 cm, initial after operation, it increased to 34.64±3.96 cm, 36.73±3.87 cm at 1 year, and 37.28±4.36 cm at the last follow-up; preoperative T 1-T 12 height was 17.38±2.76 cm. increased to 19.39±2.86 cm after the initial operation, 21.77±2.71 cm at 1 year, and 21.91±2.74 cm at the last follow-up; Preoperative coronal balance was 1.52±0.73 cm, and decreased to 0.87±0.38 cm after the initial operation, 0.81±0.38 cm at 1 year, and 0.77±0.37 cm at the last follow-up; preoperative sagittal balance was 1.94±0.78 cm, and 1.42±0.56 cm after operation, 1.28±0.55 cm at 1 year, and 1.26±0.57 cm at the last follow-up; The preoperative apical vertebra offset was 4.33±1.85 cm, and 2.16±1.47 cm after the initial operation, 1.63±1.17 cm at 1 year, and 1.61±1.23 cm at the last follow-up; SAL increased from preoperative 0.88±0.05 to 0.94±0.03 postoperatively, and 0.96±0.01 at 1-year follow-up, and 0.97±0.01 at the last follow-up. The differences between the above indicators before and after surgery were statistically significant ( P<0.05); there was a statistically significant difference in SAL between the 1-year follow-up and the last follow-up ( t=3.80, P=0.001), and other indicators were not statistically significant. Among the 20 cases, there were 5 cases of postoperative complications, including 2 cases of pedicle screw loosening and displacement, 2 cases of transverse process hook decoupling, and 1 case of proximal junctional kyphosis (PJK). The complication rate was 25% (5/20), all of them underwent revision treatment, and the prognosis was good after timely treatment. Conclusion:The dual growth bar technique can effectively control the progression of EOS deformity, preserve the longitudinal growth potential of the spine, and buy time for the development of the thorax in children, which has high safety.
10. Observation and analysis of 20 facial morphological characteristics of Xibo ethnicity adults in Chabchar in Xinjiang
Yan-Li HOU ; Qing-Yuan ZHANG ; Rong-Zhi LIU ; Guo-Chang XU ; Ming-Zhu XU ; You-Feng WEN
Acta Anatomica Sinica 2020;51(3):437-441
Objective To investigate the 20 facial morphological characteristics of Xibo ethnicity adults in Chabchar in Xinjiang, and to explore the causes of the features, so as to provide basic data for the basic study of minority anthropology. Methods A random sampling method was used to determine 677 Xibo ethnicity adults, and 20 facial morphologyical characteristics was measured and statistically analyzed. Results In terms of age groups, a total of 3 indicators which were opening height of eyeslits(