1.Hepatocyte growth factor in combination with interleukin-6 to induce the in vitro differentiation of mesenchymal stem cells isolated from human umbilical cord blood into hepatocyte-like cells
Hong YUAN ; Yuzhuo WU ; Hong CHEN
Chinese Journal of Tissue Engineering Research 2007;0(12):-
AIM: Mesenchymal stem cells (MSCs) isolated from human umbilical cord blood can differentiate into hepatocyte-like cells under a suitable induction condition. The optimal condition is still unclear. This study investigated the feasibility of differentiation of MSCs isolated from human umbilical cord blood into hepatocyte-like cells with hepatocyte growth factor (HGF) and interleukin-6 (IL-6) in vitro to find a new cell source for live tissue engineering. METHODS: The experiment was performed from March to September 2007 at the Central Laboratory of First Hospital of Lanzhou University. ①Three cases of umbilical cord blood were collected from full-term pregnant women. Pregnant women had signed an informed consent. The experiment was approved by Hospital’s Ethical Committee. ②MSCs from umbilical cord blood were separated by density gradient centrifugation and adherent method. Cell surface molecule was measured by flow cytometry. Third passage of cells were divided into four groups and cultured in DMEM medium with HGF (HGF group), IL-6 (IL-6 group), HGF+IL-6 (HGF+IL-6 group) or no growth factor (control group). ③The characteristics of proliferation and growth of MSCs were studied by microscopy and methyl thiazolyl tetrazolium (MTT). The phenotypes of MSCs were identified by flow cytometry and immunohistochemical method. Albumin levels in culture supernatants were determined with enzyme-labeled immunosorbent assay (ELISA). RESULTS: ①Growth and division of adherent cells obtained from human umbilical cord blood were good. The shape of MSCs changed into triangle, polygonal or round on days 21-28 in induction groups. ②Positive staining reaction for alpha fetoprotein (AFP) on day 7, for CK18 on days 21, 28 after induction. Albumin production by induced MSCs increased in a time-dependent manner. The positive percents of every hepatocytic marker were higher in the HGF+IL-6 group than in the IL-6 group and HGF group (P
2.Hepatocyte growth factor and epidermal growth factor induce mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells in vitro
Yuzhuo WU ; Yuting ZHANG ; Xiaorong MAO ; Hong CHEN
Journal of Third Military Medical University 2003;0(16):-
Objective To explore the possibility of differentiation of mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells induced by EGF and HGF and to lay basis for transplanted autologous bone marrow MSCs in treatment of liver disease at terminal stage.Methods Bone marrow cells were obtained from volunteers with liver cirrhosis.MSCs were separated by density gradient centrifugation and were cultured through adhere culture.MSCs were cultured in DMEM medium with HGF,EGF,HGF+EGF or no growth factor.The phenotypes of MSCs were identified by flow cytometry,immunohistochemistry,and Albumin levels in culture supernatants were determined by ELISA.Results Growth and division of adherent cells obtained from the patients with hepatocirrhosis were good and the phenotypes of MSCs were CD29 positive and CD34 negative.The shape of MSCs changed from long fusiform to polygonal or round on 21th-28th days in grow factor induced groups.Immunocytochemical analysis for CK18 and AFP showed positive staining reaction for AFP on 7th day,for CK18 on 21st and 28th day in grow factor induced groups with MSCs-induced Alb production increasing in a time-dependent manner.No markers of hepatocyte linear cells were detected in no growth factor induction group.Conclusion Both HGF and EGF can induce mesenchymal stem cells to differentiate hepatocyte-like cells alone or coordinately.
3.Clinical observation of preventive use of antibiotic inguinal hernial repair without tension and its significance
Junzhong SHI ; Jianbin ZHUANG ; Huijun SUN ; Yuzhuo CHEN ; Weiliang SONG ; Xiangchao MENG
Journal of Jilin University(Medicine Edition) 2017;43(3):622-625
Objective:To explore the necessity of the preventive use of antibiotics and the effects of age and operation time on the efficacy of inguinal hernia repair without tension,and to elucidate the clinical significance of the preventive application of prophylactic antibiotics in inguinal hernia repair without tension.Methods:A total of 228 patients with inguinal hernia repair without tension were selected,amomg them 42 cases with high infection factors were treated with antibiotics (treated group),and 186 cases were not treated with antibiotics(untreated group) during the preoperative period.The prophylactic antibiotics were given 30 min before surgery,and the conventional dose was not used more than 48 h after surgery.All the cases were treated with artificial repair materials for the procedure of inguinal hernial repair without tension.The age,highest body temperature,white blood cell count,operation time,hospitalization time,and postoperative body temperature of all the 228 cases were recorded and analyzed statistically.Results:The preoperative and postoperative white blood cell counts had significant differences between the patients<60 years and the patients≥60 years in untreated group (P<0.05);there were no significant differences in the operation time,hospitalization time and body temperature between the patients<60 years and the patients≥60 years in untreated group (P>0.05).Compared with the patients with the operation time>90 min,the white blood cell count and hospitalization time of the patients with the operation time ≤90 min were increased (P<0.05).There were no significant differences of the body temperature and age between the patients with the operation time >90 min and the patients with the operation time≤90 min (P>0.05).The white blood cell count,operation time,hospitalization time and postoperative body temperature of the patients between treated group and untreated group had no significant differences (P>0.05).Conclusion:The use of antibiotics in the high-risk patients and non-use of antibiotics in the majority of elective inguinal hernia repair without tension can ensure the safe and performability of the patients.
4.Establishment of loop-mediated isothermal amplification technique for rapid detection of NDM-1 gene.
Yuanyi ZHANG ; Na WU ; Baoli ZHU ; Lei CHEN ; Yuzhuo ZHU
Chinese Journal of Biotechnology 2011;27(8):1232-1238
We established a rapid detection method of New Delhi Metallo-beta-Lactamase Gene (NDM-1) based on Loop-mediated Isothermal Amplification (LAMP). With the application of LAMP, we designed four sets of LAMP premiers, using NDM-1 gene as the target sequence, and selected the set of optimal primers. Meanwhile, we established optimal reaction systems and conditions to carry out the sensitivity and specificity experiments. The experiment results showed that the whole detection process took only one hour and could be observed visually. In the experiment of sensitivity, NDM-1 gene had a detection limit of 6 copies in each reaction. In the experiment of specificity, we detected NDM-1 gene in 4 pathogen strains (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae), and the total DNA from intestinal microbes and the total DNA from soil microbes. We had not detected the amplification reactions. The detection method established could rapidly detect NDM-1 gene and visualize the experiment result. The method is easy to operate and has high sensitivity and specificity and thus has great application value in basic research laboratories, emergent detection and spot detection.
Bacteria
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enzymology
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genetics
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Bacteriological Techniques
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methods
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DNA, Bacterial
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genetics
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Escherichia coli
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enzymology
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genetics
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Klebsiella pneumoniae
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enzymology
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genetics
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Nucleic Acid Amplification Techniques
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methods
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Sensitivity and Specificity
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Staphylococcus aureus
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enzymology
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genetics
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Streptococcus pneumoniae
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enzymology
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genetics
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beta-Lactamases
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genetics
5.Recent therapeutic drug progress on neuroendocrine prostate cancer
Zheng CHEN ; Liwei WEI ; Yuzhuo WANG ; Yumin ZHUO
Chinese Journal of Urology 2020;41(9):709-712
Neuroendocrine prostate cancer(NEPC) is a prostate cancer subtype with a very high degree of malignancy and a special molecular phenotype.NEPC is not sensitive to endocrine therapy, and there are currently no specific drugs, so there is a lack of effective clinical treatment.New advances in NEPC therapeutic include chemical therapy, targeted drug therapy based on molecular phenotype and other non-targeted drug therapy. This article summarizes the current treatment methods, pharmaceutical, and clinical research results for NEPC, aiming to deepen clinicians' more comprehensive understanding of NEPC patients' treatment strategies.
6.Investigation and analysis of somatic symptom disorder and anxiety and depression in patients with myocardial bridge
Bingxu CHEN ; Yixuan LI ; Yuzhuo LIU ; Xiaoyu GUAN ; Meng JIANG ; Jialiang MAO
Chinese Journal of Behavioral Medicine and Brain Science 2020;29(2):130-135
Objective:To investigate and analyze the somatic symptom disorder, anxiety and depression in patients with myocardial bridge.Methods:A total of 276 patients with myocardial bridge diagnosed by coronary angiography (CAG) were enrolled in the department of cardiology of Renji hospital in Shanghai from June to December in 2016. There were 151 cases of simple myocardial bridge (no coronary stenosis or coronary artery stenosis <30%) and 125 cases of complex myocardial bridge (combined with coronary stenosis >30%). A total of 1067 patients with myocardial bridge without coronary angiography were collected at the same time. Self-rating somatic symptom scale (SSS), generalized anxiety disorder (GAD-7) and patient health questionnaire (PHQ -9) were given to these patients during hospitalization. At the same time, somatic symptoms disorder and anxiety and depression in the myocardial bridge group and non-myocardial bridge group were compared.Results:The prevalence of somatic symptom disorder in patients with myocardial bridge was higher than that in non-myocardial bridge patients (35.86% vs 28.30%, P<0.05). There was significant correlation between somatic symptom disorder and depression and anxiety, with correlation coefficients of 0.629 and 0.565, respectively. The prevalence of depression and anxiety in myocardial bridge patients was higher than that in non-myocardial bridge patients (depression: 23.91% vs 22.11%. P=0.467; anxiety: 17.02% vs 14.15%, P=0.22), but there was no statistical difference. For male patients or female patients, the prevalence of somatic symptom disorder, depression and anxiety in the simple myocardial bridge patients were higher than those in the complex myocardial bridge patients, but there was no statistical difference. The most common non-specific somatic symptoms disorder in patients with myocardial bridge were fatigue (64.5%), followed by sleep disorders (63.8%) and decreased attention (63.0%). Conclusion:The somatic symptom disorder in patients with myocardial bridge is significantly higher than that in non-myocardial bridge group. Especially for patients with myocardial bridge with non-specific somatic symptoms, early identification of somatic symptoms disorder of myocardial bridge patients will be beneficial to proper clinical invitation.
7.A novel draw-bar skin stretcher for repair of full-thickness skin defects
Yuzhuo HAN ; Yonghua CHEN ; Dong LIU ; Qingshan GUO ; Siheng DU ; Siru ZHOU ; Lianyang ZHANG ; Yang LI
Chinese Journal of Orthopaedic Trauma 2021;23(7):627-631
Objective:To evaluate a new type of draw-bar skin stretcher in repair of full-thickness skin defects.Methods:From May 2015 to January 2019, 52 patients with full-thickness skin defects were repaired with a new type of draw-bar skin stretcher at Daping Hospital, Army Medical University. They were 40 males and 12 females, aged from 4 to 61 years (average, 37.1 years). Their skin was stretched for primary wound closure. When primary wound closure failed, skin stretching was performed again to close the wound depending on the wound condition. When the Pinch test was negative after skin stretching, the wound was sutured directly. In cases of positive Pinch test, a skin graft or flap was used to repair the remaining wound. At 12 months after surgery, scar contracture and size of skin graft or flap were observed and wound healing after skin stretching was evaluated in comparison with the original wound.Results:After skin stretching, one-stage wound closure was achieved in 36 cases and multi-stage wound closure in 8 cases; of the remaining 8 cases, 2 were repaired by skin graft and 6 by skin flap after their wounds were reduced by skin stretching. In one-stage closed wounds, infection occurred in 3 cases and marginal necrosis in 5 cases; in the wounds repaired by skin graft or flap, no infection or necrosis was observed. The 12-month follow-up for all the patients showed fine healing of all the wounds after one-stage or multi-stage closure, linear scar, absence of scar contracture, and smaller wound sizes than the original ones after skin graft or flap repair.Conclusions:Skin stretching using our new type of draw-bar skin stretcher is an effective treatment for skin wounds. It can replace traditional skin grafting and flap surgery in some cases, but its indications should be strictly followed to avoid related complications.
8. Effect of papaverine on wound healing of full-thickness skin defect after skin stretching
Yuzhuo HAN ; Yang LI ; Dong LIU ; Qingshan GUO ; Yonghua CHEN ; Lianyang ZHANG
Chinese Journal of Trauma 2019;35(9):853-859
Objective:
To investigate the effect of papaverine on wound healing of full-thickness skin defect after skin stretching in swine.
Methods:
Eight Bama pigs were prepared. Standard full-thickness skin defect for 7 cm×7 cm was produced in the middle part of each pig's forelimb, and skin traction was used to close the wound. According to random number table method, the pigs were divided into experiment group and control group, with four pigs in each group. Percutaneous oxygen partial pressure (TcPO2) was compared preoperatively and postoperatively. Wound healing clinical score was evaluated four and 10 days after operation. Weidner counting method was employed to determine the microvascular density (MVD) of the wound edge tissue 0 and 10 days after operation. Immunohistochemistry was used to observe the expression of hypoxia inducible factor 1-α (HIF- 1α) and vascular endothelial growth factor (VEGF) 0, 4 and 10 days after operation.
Results:
There was no statistically significant difference in TcPO2 between the experiment group [(55.1±5.4)mmHg] and the control group [(54.7±5.9)mmHg] before skin stretching (
9.Effects of electroacupuncture at "Weizhong" (BL 40) on morphology and expression of CK and IL-17 in rats with bupivacaine-induced multifidus muscle injury.
Dehui ZOU ; Yupei CHEN ; Tong LIU ; Zongxiao LU ; Jun YAN ; Dongli CHEN ; Yue XU ; Jiayi ZHANG ; Yuzhuo BAI ; Li ZHANG ; Zejun HUO
Chinese Acupuncture & Moxibustion 2017;37(9):971-976
OBJECTIVETo observe effects of electroacupuncture (EA) at "Weizhong" (BL 40) on morphology and expression of creatine kinase (CK) and interleukin-17 (IL-17) in rats with bupivacaine-induced multifidus muscle injury.
METHODSA total of 32 male SD rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 8 rats in each one. The rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine to establish the model of multifidus muscle injury; the rats in the control group were injected with 0.9% sodium chloride solution. The rats in the Weizhong group and Shenshu group were treated with EA (2 Hz/10 Hz in frequency, 1~2 mA in intensity) at "Weizhong" (BL 40) and "Shenshu" (BL 23), 20 min per treatment. No treatment was given in the control group and model group. After 14-day treatment of EA, the inflammatory cell count, scar tissues area and muscle fiber cross sectional area of multifidus muscle were observed with HE and Masson staining method. The activity of CK and serum content of IL-17 were test with enzyme-linked immunosorbent assay (ELISA) method; the expression of IL-17 in multifidus muscle was measured with immunohistochcmical method.
RESULTSAfter intervention, the inflammatory cell count and scar tissues area in the model group, Weizhong group and Shenshu group were higher than those in the control group (all<0.01), but the muscle fiber cross sectional area was significantly reduced (all<0.01); the inflammatory cell count and scar tissues area in the Weizhong group and Shenshu group were lower than those in the model group (all<0.01), and the muscle fiber cross sectional area was significantly increased (<0.01,<0.05). After intervention, the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the model group, Weizhong group and Shenshu group were higher than those in the control group (all<0.01); the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the Weizhong group and Shenshu group were lower than those in the model group (<0.01,<0.05); compared with the Shenshu group, the down-regulation of IL-17 was more obvisous in the Weizhong group (<0.01).
CONCLUSIONEA at "Weizhong" (BL 40) can down-regulate the overexpression of serum CK and IL-17, alleviate inflammation reaction and improve the repair of multifidus muscle.
10.The traditional Chinese medicines treat chronic heart failure and their main bioactive constituents and mechanisms.
Jie CHEN ; Xiaohong WEI ; Qian ZHANG ; Yuzhuo WU ; Guiyang XIA ; Huan XIA ; Lingyan WANG ; Hongcai SHANG ; Sheng LIN
Acta Pharmaceutica Sinica B 2023;13(5):1919-1955
Chronic heart failure (CHF) is a severe public health problem with increasing morbidity and mortality, any treatment targeting a single session is insufficient to tackle this. CHF is characterized by reduced cardiac output resulting from neurohumoral dysregulation and cardiac remodeling, which might be related to oxidative stress, inflammation, endoplasmic reticulum stress, apoptosis, autophagy, mitochondrial function, and angiogenesis. These molecular mechanisms interact with each other through crosstalk. Historically, Chinese medicinal herbs have been widely applied in the treatment of CHF, and therapeutic effects of Chinese medicinal herbs and their ingredients have been scientifically confirmed over the past decades. Traditional Chinese medicine (TCM) with multiple components can confront the different pathogenesis of CHF through multiple targets. This review analyzes commonly used TCM patent drugs and TCM decoctions that are applicable to different stages of CHF based on clinical trials. Diverse bioactive ingredients in Chinese medicinal herbs have been found to treat CHF via multiple molecular mechanisms. This review comprehensively covers the key works on the effects and underlying mechanisms of TCM, herbal ingredients and synergistic effects of constituent compatibility in treating CHF, providing additional ideas to address this threat.