1.Effect of Complanatoside A on the apoptosis of articular chondrocytes
Lu YIN ; Chuanfeng JIANG ; Junjie CHEN ; Ming YI ; Zihe WANG ; Houyin SHI ; Guoyou WANG ; Huarui SHEN
Chinese Journal of Tissue Engineering Research 2025;29(8):1541-1547
BACKGROUND:Chondrocyte apoptosis is an important factor in the development of osteoarthritis,and Complanatoside A has a flavonoid effect,which can inhibit apoptosis of various cells,but its effect on chondrocyte apoptosis and the mechanism of action are not clear. OBJECTIVE:To investigate the intrinsic association and mechanism of Complanatoside A in chondrocyte apoptosis based on the Wnt/β-catenin signaling pathway. METHODS:(1)The cartilage tissues of the femur and tibia transected during knee arthroplasty were collected,and chondrocytes were isolated,cultured in vitro,and identified.(2)Cell counting kit-8 was used to detect the optimal intervention concentration of Complanatoside A in the concentration range of 0-160 μmol/L.(3)Chondrocytes were divided into blank group,sodium nitroprusside(1.5 mmol/L)-induced group,and sodium nitroprusside(1.5 mmol/L)+Complanatoside A(5 μmol/L)group.The viability and apoptosis rate of the cells in each group were detected by cell counting kit-8 and flow cytometry.The expression of type Ⅱ collagen and SOX9 was detected by immunofluorescence staining.The expression of apoptosis-related proteins and Wnt/β-catenin pathway proteins was detected by western blot assay. RESULTS AND CONCLUSION:The cells extracted in vitro were cultured and stained,and were clearly identified as chondrocytes.Complanatoside A had no obvious cytotoxicity to chondrocytes in the concentration range of 0-80 μmol/L,and significantly improved the chondrocyte viability in the concentration range of 2.5-10 μmol/L,especially when the concentration was 5 μmol/L.The apoptotic rate of chondrocytes was higher in the sodium nitroprusside-induced group than the blank control group,while the apoptotic rate was lower in the sodium nitroprusside+Complanatoside A group than the sodium nitroprusside-induced group.The fluorescence intensity of type Ⅱ collagen and SOX9 in chondrocytes was weaker in the sodium nitroprusside-induced group than the blank control group,while the fluorescence intensity of type Ⅱ collagen and SOX9 in the sodium nitroprusside+Complanatoside A group was higher than that of the sodium nitroprusside-induced group.In the sodium nitroprusside-induced group,the protein expression of Bax,Caspase-3,matrix metalloproteinase 13,Wnt3a,Wnt5a and β-catenin was higher than that of the blank control group,while the protein expression of Bcl-2 was lower than that of the blank control group.In the sodium nitroprusside+Complanatoside A group,except for the protein expression of Bcl-2 which was higher than that of the sodium nitroprusside-induced group,the expression of the other aforementioned proteins was lower than that of the sodium nitroprusside-induced group.To conclude,Complanatoside A has a certain inhibitory effect on chondrocyte apoptosis,which could regulate apoptosis-related proteins and promote the expression of chondrocyte regulatory factors,and presumably might play a role through inhibiting the Wnt/β-catenin signaling pathway.
2.Expressions of survivin, p53 and human epidermal growth factor receptor 2 in breast cancer tissues and their prognostic value
Hua CHENG ; Guoyou DONG ; Xiaoye WANG ; Yunxiao ZHANG ; Zhiying LIU
Cancer Research and Clinic 2023;35(1):1-6
Objective:To investigate the expressions of apoptosis-related factors survivin, p53 and human epidermal growth factor receptor 2 (HER2) in breast cancer tissues and their prognostic value.Methods:A total of 131 patients undergoing radical mastectomy for breast cancer who were admitted to Tangshan Maternal and Child Health Care Hospital from February 2015 to January 2019 were selected as the research subjects. During the operation, the cancer tissues and adjacent tissues (normal tissues >3 cm from the tumor margin) were collected from the patients. Expressions of survivin, p53 and HER2 in cancer tissues and adjacent tissues of patients were detected by using immunohistochemistry. The prognoses of patients were recorded after the follow-up for 3 years; the recurrence, metastasis and death treated as the poor prognosis, the rest prognoses of patients were treated as the good prognosis group. The difference of clinicopathological characteristics between the poor prognosis group and the good prognosis group was compared. Multivariate logistic regression was used to analyze risk factors for prognosis of breast cancer patients. The result of prognosis of breast cancer was taken as the golden standard. The receiver operating characteristic (ROC) curve was used to analyze the value of survivin pasitive, p53 pasitive, HER2 pasitive alone, the combination of both and the combination of the there in the judgement of poor prognosis of breast cancer.Results:The positive expression rates of survivin [49.6% (65/131) vs. 7.6% (10/131)], p53 [60.3% (79/131) vs. 13.0% (17/131)] and HER2 [79.4% (104/131) vs. 16.8% (22/131)] in cancer tissues were higher than those in adjacent tissues (all P<0.001). A total of 131 breast cancer patients were followed up for 3 years without any loss of follow-up, and the follow-up rate was 100%. Within the follow-up for 3 years, there were 15 (11.5%) cases of recurrence, 8 (6.1%) cases of metastasis, and 10 (7.6%) cases of death, the incidence of poor prognosis was 25.2% (33/131); and the remaining 98 cases had good prognosis. The proportions of patients with TNM stage Ⅲ, lymph node metastasis, poorly differentiated histology, tumor diameter ≥3 cm, survivin, p53, and HER2 positive expressions in the poor prognosis group were higher than those in the good prognosis group (all P<0.05). Multivariate logistic regression analysis showed that TNM stage Ⅲ [ OR = 5.323 (95% CI 2.190-12.936)], lymph node metastasis [ OR = 4.773 (95% CI 1.964-11.600)], tumor diameter ≥3 cm [ OR = 3.582(95% CI 1.474-8.706)], positive survivin [ OR = 2.740 (95% CI 1.127-6.659)], positive p53 [ OR = 3.271 (95% CI 1.346-7.949)], and positive HER2 [ OR = 3.873 (95% CI 1.594-9.412)] were independent risk factors for prognosis of breast cancer (all P<0.001). The ROC curve results showed that the area under the curve (AUC) values of survivin positive, p53 positive,HER2 positive, and the combination of any two were more than 0.80 (all P<0.001); the AUC of the combination of the three was 0.944 (95% CI 0.890-0.977) ( P<0.001). Conclusions:The expressions of survivin, p53, and HER2 are highly expressed in breast cancer tissues. The expressions of the three can be used to judge the prognosis of breast cancer patients, and the combination of the three has a higher judgement value.
3.A novel inhibitor of N 6-methyladenosine demethylase FTO induces mRNA methylation and shows anti-cancer activities.
Guoyou XIE ; Xu-Nian WU ; Yuyi LING ; Yalan RUI ; Deyan WU ; Jiawang ZHOU ; Jiexin LI ; Shuibin LIN ; Qin PENG ; Zigang LI ; Hongsheng WANG ; Hai-Bin LUO
Acta Pharmaceutica Sinica B 2022;12(2):853-866
N 6-methyladenosine (m6A) modification is critical for mRNA splicing, nuclear export, stability and translation. Fat mass and obesity-associated protein (FTO), the first identified m6A demethylase, is critical for cancer progression. Herein, we developed small-molecule inhibitors of FTO by virtual screening, structural optimization, and bioassay. As a result, two FTO inhibitors namely 18077 and 18097 were identified, which can selectively inhibit demethylase activity of FTO. Specifically, 18097 bound to the active site of FTO and then inhibited cell cycle process and migration of cancer cells. In addition, 18097 reprogrammed the epi-transcriptome of breast cancer cells, particularly for genes related to P53 pathway. 18097 increased the abundance of m6A modification of suppressor of cytokine signaling 1 (SOCS1) mRNA, which recruited IGF2BP1 to increase mRNA stability of SOCS1 and subsequently activated the P53 signaling pathway. Further, 18097 suppressed cellular lipogenesis via downregulation of peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and C/EBPβ. Animal studies confirmed that 18097 can significantly suppress in vivo growth and lung colonization of breast cancer cells. Collectively, we identified that FTO can work as a potential drug target and the small-molecule inhibitor 18097 can serve as a potential agent against breast cancer.
4.Comparison on Differences of Volatile Components in Chuanxiong Chatiaosan with Different Compatibility of Chuanxiong Rhizoma Decoction Pieces
Yun WANG ; Guoyou WANG ; Yinghao ZHENG ; Yanan SONG ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(16):180-187
ObjectiveTaking Chuanxiong Chatiaosan prescription as the carrier, by comparing the differences of volatile components in Chuanxiong Rhizoma with single decoction pieces and compatible prescription of different decoction pieces, the differences of material basic connotation of different formulations of Chuanxiong Chatiaosan were revealed from the aspects of processing (raw and wine-processed products), compound compatibility and dosage form (powder and decoction). MethodThe volatile oil was extracted from different decoction pieces of Chuanxiong Rhizoma, Chuanxiong Chatiaosan and its decoction with different decoction pieces of Chuanxiong Rhizoma by steam distillation, the main components and their relative contents were identified by gas chromatography-mass spectrometry (GC-MS). ResultA total of 25 volatile components were identified from different processed products of Chuanxiong Rhizoma, including 11 monoterpenoids, 4 phenols, 3 sesquiterpenoids, 3 phthalides, 2 ketones and 2 olefins, the contents of α-pinene, β-pinene, 3-butylphthalide and others increased after the raw products was processed with wine. A total of 85 constituents were identified from Chuanxiong Chatiaosan with different decoction pieces, including 31 monoterpenoids, 23 sesquiterpenoids, 5 alcohols, 5 aldehydes, 4 phenols, 4 phthalides, 3 ethers, 3 ketones, 1 olefin, 1 organic acid, 2 esters and 3 other compounds. A total of 22 components, including 9 sesquiterpenoids, 3 phthalides, 2 phenols, 6 monoterpenoids, 1 aldehyde and 1 alkane, were identified from the decoction of Chuanxiong Chatiaosan with different processed products. ConclusionThere was no significant difference in the composition between raw products and wine-processed products of Chuanxiong Rhizoma either in single decoction pieces or in compatibility prescription, but the relative content changed to some extent, and the wine-processed products was the most obvious. There was a great difference in the composition of volatile components between the Chuanxiong Chatiaosan and its decoction. The volatile components, such as isopulegol, isocalamendiol and safrole, were not found in the decoction. Components in Chuanxiong Rhizoma processed with wine will change with the addition of yellow rice wine, and volatile components can reflect the difference between decoction pieces and prescriptions of the wine-processed products.
5.Study on the Mechanism of Enhancement Effects of Astragalus complanatus Polysaccharides on the Proliferation of Meniscal Fibrochondrocyte Cells in Rabbits
Huarui SHEN ; Liang AO ; Lisheng WANG ; Guoyou WANG ; Qi HAO ; Ting LI
China Pharmacy 2020;31(9):1097-1102
OBJECTIVE:To study the mechanism of enhancement effects of Astragalus complanatus polysaccharides(ACP) on the proliferation of meniscal fibrochondrocytes cells in rabbits. METHODS :The meniscal fibrochondrocytes cells were isolated from 1-month-old New Zealand white rabbits. The meniscal fibrochondrocytes cells were divided into normal control group (PBS), positive control group (glucosamine sulfate ,10 mg/mL)and ACP high-dose,medium-dose and low-dose groups (40,20,10 mg/mL). The morphology of meniscal fibrochondrocytes cells were observed under microscope. Cell proliferation rate was detected by MTT assay. Cell cycle was observed with flow cytometry. ELISA assay was used to detect relative expression of medium collagen type Ⅱ(Col Ⅱ)and alkaline phosphatase protein (ALP)in meniscal fibrochondrocytes cells. RT-qPCR and Western blotting assay were adopted to detect mRNA and protein expression of transforming growth factor β 1 (TGF-β 1) and bone morphogenetic protein 2(BMP-2). RESULTS :After cultured for 72 h,meniscal fibrochondrocytes cells were fused into a single layer,and most of them were slender type in appearance. Compared with normal control group ,the proliferation rate of meniscal fibrochondrocytes cells and the percentage of cells at G 1/G0 phase were decreased significantly in positive control group and ACP high-dose,medium-dose and low-dose groups (P<0.05);the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP,mRNA and protein expression of TGF-β1 and BMP- 2 were increased significantly (P<0.05). Compared with positive control group,inhibitory rate of meniscal fibrochondrocytes cells proliferation and the percentage of cells at G 1/G0 phase were decreased significantly in ACP high-dose group (P<0.05),while the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP , mRNA and protein expression of TGF-β1 and BMP- 2 were increased significantly (P<0.05). The inhibitory proliferation rate of meniscal fibrochondrocytes cells and the percentage of cells at G 1/G0 phase were increased significantly in ACP low-dose group (P< 0.05),while the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP ,mRNA and protein expression of TGF-β1 and BMP- 2 were decreased significantly (P<0.05). There was no statistical significance in above indexes of ACP medium-dose group. CONCLUSIONS :ACP can promote the proliferation of meniscal fibrochondrocytes cells ,reduce the percentage of cells at G1/G0 phase,promote cell transformation to S phase ;the mechanism of which may be related to up-regulating TGF-β1,BMP-2 mRNA and protein expression ,promoting Col Ⅱ and ALP protein expression enhancement.
6.Double-row suture anchors under arthroscopy for avulsion-type greater tuberosity fractures (Mutch type I)
Gang LIU ; Lei ZHANG ; Guoyou WANG ; Xin ZHOU ; Tao ZHANG ; Taiyuan GUAN ; Xiaoguang GUO ; Shijie FU
Chinese Journal of Tissue Engineering Research 2017;21(19):3005-3010
BACKGROUND: Greater tuberosity of humerus, as the attachment point of rotator cuff, plays a vital role in shoulder.Neer and AO classification for proximal humeral fractures have been accepted extensively. However, the classification for single greater tuberosity fractures of the proximal humerus is little reported, and its fixation method remains controversial.OBJECTIVE: To explore the curative efficacy of the double-row suture anchors under arthroscopy for avulsion-type greater tuberosity fractures (Mutch type Ⅰ) METHODS: Clinical data of 24 patients with avulsion-type greater tuberosity fractures (Mutch type Ⅰ) undergoing double-row suture anchors under arthroscopy were analyzed retrospectively. The displacement distance of the greater tuberosity of humerus was measured on CT before and after fixation. The shoulder joint was evaluated by Constant-Murley Score and the University of California, Los Angeles score at baseline, 1, 3, 6 and 12 months postoperatively. Besides, the visual analogue scale scores were detected at baseline and 12 months postoperatively.RESULTS AND CONCLUSION: (1) Double-row suture anchors effectively improved the displacement of greater tuberosity and rotator cuff injury. (2) The patients accompanied by rotator cuff injury accounted for 79.16%. (3) The Constant-Murley Score and the University of California, Los Angeles scores were significantly improved at each time point (P < 0.05). The visual analogue scale scores after fixation were significantly superior to those before fixation (P < 0.05). (4) These results suggest that the double-row suture anchors under arthroscopy can effectively improve the displacement of greater tuberosity and alleviate the pain. Moreover, it is conductive for early recovery of the shoulder function with little trauma, so it is a good choice for avulsion-type greater tuberosity fractures (Mutch type Ⅰ).
7.Anterior cruciate ligament reconstruction with autologous ipsilateral 1/2 peroneus longus tendon
Lei ZHANG ; Xin ZHOU ; Ji QI ; Taiyuan GUAN ; Lijun XIE ; Guoyou WANG ; Shijie FU ; Yikai LI
Chinese Journal of Tissue Engineering Research 2017;21(24):3815-3820
BACKGROUND:Anterior cruciate ligament (ACL) injury is a commonly sport-induced knee joint injury that does serious harm to the knee stability. ACL reconstruction is a commonly used treatment method, but researches on 1/2 peroneus longus tendon (PLT) graft are rarely reported. OBJECTIVE: To investigate the clinical outcomes of removing the autologous ipsilateral 1/2 PLT under arthroscopy for ACL reconstruction. METHODS:106 patients with complete ACL rupture in the Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University from December 2010 to December 2014 were enrolled, and autologous ipsilateral 1/2 PLT was removed under arthroscopy for ACL reconstruction. At baseline, 3, 6 and 12 months postoperatively, the knee stability was evaluated manually through the anterior drawer test, Lachman test, and pivot-shift test, and the knee function was evaluated by Tegner activity scale, Lysholm and International Knee Documentation Committee scores. RESULTS AND CONCLUSION: Postoperative anterior drawer test, Lachman test, and pivot-shift test tests were negative in all patients. In terms of Tegner activity scale, Lysholm and International Knee Documentation Committee scores, there were significant differences at baseline and postoperative 3 months as compared with postoperative 6 months (P < 0.05); the scores at baseline and postoperative 3 months showed significant differences compared with 12 months postoperatively (P < 0.05); the scores showed no significant difference between 6 and 12 months postoperatively (P > 0.05). These results indicate that autologous ipsilateral 1/2 PLT is a good choice for ACL reconstruction under arthroscopy, achieving rapid and satisfactory functional recovery of the knee joint, which is not only minimally invasive and easy to operate, but also exhibits good therapeutic efficacy.
8.Development of an Independent Heater for Ventilator Tube.
Chinese Journal of Medical Instrumentation 2016;40(1):44-46
Nowadays, normal humidifier is used to heat and humidify the gas before sending to ventilator tube. A new type of ventilator which offers both breathing tube with heater and humidifier is incorporate. In the light of this, patients already bought ventilator still confront this problem. Therefore, this paper mainly introduces a new manufactural method which is controlling the temperature and humidity of gas sent by breathing machine online by a temperature controller which consist of Silica gel hotline and microcomputer. As a matter of fact, the controller is adaptable in various types of breathing tube and can accurately control the humidity and temperature of gas sent into bodies.
Equipment Design
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Humans
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Humidity
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Temperature
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Ventilators, Mechanical
9.Changes in miRNAs expression profiling in the liver of a rat model after reduced-size liver transplantation
Hongqiang GAO ; Zhiqiang LI ; Haifu WANG ; Guoyou XUE ; Jing LIU ; Gang CHEN ; Li LI
Chinese Journal of Tissue Engineering Research 2016;20(5):712-717
BACKGROUND:Liver regeneration is the key factor influencing the prognosis of living donor liver transplantation. There has not been the research on special miRNA of liver regeneration after living donor liver transplantation. OBJECTIVE:To analyze the variation of miRNAs expression profile after rat reduced-size liver transplantation at certain time point, select and verify target miRNA which can provide targeting intervention strategies in liver regeneration after rat reduced-size liver transplantation and provide theoretical evidence for liver regeneration after living donor liver transplantation.METHODS:The reduced-size liver transplantation models were established. miRNAs microarray was used to detect miRNA expression. In differentialy expressed microRNAs, real-time quantitative PCR was utilized to detect target miRNAs. The credibility of miRNAs microarray results was verified. RESULTS AND CONCLUSION:Compared with rat liver tissue in the sham operation group, 11 miRNAs up-regulated in reduced-size liver transplantation, including let-7b-5p, let-7c-5p, miR-101a-3p, miR-103-3p, miR-130a-3p, miR-142-5p, miR-186-5p, miR-199a-3p, miR-21-5p, 221-3p and miR-34a-5p. Four miRNAs were down-regulated, including miR-26b-5p, miR-150-5p, miR-19a-3p and rno-miR-146-5p. PCR test further verified that miR-221-3p and miR-199a-3p expression changes approximated the chip results at 24, 48 hours and 1 week, indicating that results of miRNA microarray were believable. These results verified that it exists variation of miRNAs expression profile after rat reduced-size liver transplantation, which picked out and verified the target miRNAs.
10.Curative efficacy analysis for metronidazole tablets combined with sophora gel in treatment of bacterial vaginitis
Xiaomei WANG ; Xuemei WANG ; Guijie QI ; Danli GAO ; Xuehui ZHANG ; Jinmei LEI ; Guoyou DONG
Chinese Journal of Biochemical Pharmaceutics 2015;37(7):128-129,132
Objective To observe the clinical efficacy of metronidazole tablets combined with sophora gel in treatment of bacterial vaginitis ( BV) . Methods Eighty-seven cases of BV patients were randomly divided into the study group(45cases) and the control group(42cases).The control group was given metronidazole tablets (400 mg, two times a day), oral for seven days, while the study group was given sophora gel (vaginal implantation) on the basis of control group.Seven days as a course of treatment.Pared the leukocyte esterase test positive rate in vaginal fluid and the recurrence rate within a year of the two groups before and after treatment.ResuIts After 7 days’treatment, the positive rate of leukocyte esterase (LE) in two groups were all significantly declined, which was more significant in the study group (P<0.05);the total effective rate of the study group was 95.6%, which was significantly higher than that of the control group 78.6%(P<0.05).The recurrence rate within a year of the study group was 6.7%, which was significantly lower than that of the control group 26.2%( P<0.05 ) .ConcIusion The therapy of metronidazole tablets combined with sophora gel in treating BV can significantly decline the LE positive rate and recurrence rate, and improve the clinical curative efficacy as well.

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