1.Quantitative analysis of transplanted effect of human amniotic epithelial cells in mice with acute liver injury
Hongwu LUO ; Quan XUN ; Xiangjun HUANG ; Feizhou HUANG
Chinese Journal of Tissue Engineering Research 2013;(32):5834-5839
BACKGROUND:There are many preliminary studies on the survival, metaptosis, and correlation characteristics of human amniotic epithelial cel s after transplanted into the animals, but there are no reports on the quantitative analysis of the transplantation effect. OBJECTIVE:To make quantitative analysis on serum biochemical function of liver and the expression of human albumin in mice received passaged human amniotic epithelial cel s transplantation in spleen. METHODS:Forty nude mice were randomly divided into four groups (n=10 in each group):hepatectomy+cel transplantation 2 weeks group, hepatectomy+cel transplantation 4 weeks group, hepatectomy+normal saline group (treated with partial hepatectomy) and hepatectomy+cel transplantation group (transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen, and the blood were col ected at 2 and 4 weeks after transplantation). The mice in the hepatectomy+normal saline group were treated with splenic injection of 0.2 mL normal saline;the cel transplantation group did not receive hepatectomy, and transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen. The histological and morphological changes of the liver and spleen in each group as wel as the expressions of serum alanine aminotransferase, aspartate aminotransferase and human serum albumin in each group were detected, and the quantitative analysis of human serum albumin expression was performed. RESULTS AND CONCLUSION: There was no obvious morphological change after human amniotic epithelial cel s transplanted into the acute liver injury mice for 4 weeks, but specific cel s could be detected by histological method. The serum levels of alanine aminotransferase, aspartate aminotransferase and human serum albumin were improved obviously, and the human albumin could be detected in serum, the level of human albumin at 4 weeks after transplantation was significantly increased than 2 weeks after transplantation. Human amniotic epithelial cel s can survive for more than 4 weeks after transplanted into the liver injury mice, and can stil express partial characteristics and functions of hepatocyte-like cel s, improve the liver function, thus treating acute liver injury.
2.Adeno-associated virus vector-medicated green fluorescent protein transfected human amniotic epithelial cells in vitro
Hongwu LUO ; Xiangjun HUANG ; Xunyang LIU ; Feizhou HUANG
Chinese Journal of Tissue Engineering Research 2010;14(10):1843-1846
BACKGROUND:Human amniotic epithelial cells(AECs)are easy to obtain and can function as ideal seed cells for cell transplantation and tissue repair.Currently,marking and tracing of human AECs remains rarely reported.OBJECTIVE:To explore the efficiency of adeno-associated virus(AAV)vector-medicated green fluorescent protein(GFP)on in vitro cultured human AECs transfection.METHODS:Human amnion samples were harvested and trypsinized to isolate human AECs.The AECs were transfected with AAV-GFP,and the transfection efficiency was detected.RESULTS AND CONCLUSION:Human AECs were successfully primary cultured and passaged in vitro.AAV-GFP-transfected AECs stably and highly expressed GFP,with a transfection efficiency of 58%.
3.Transversal inducing differentiation of human amniotic epithelial cells into hepatocyte-like cells
Hongwu LUO ; Xiangjun HUANG ; Feizhou HUANG ; Xunyang LIU
Journal of Central South University(Medical Sciences) 2011;36(6):525-531
Objective To evaluate the in vitro differentiation of human amniotic epithelial cells (hAECs ) into hepatocyte-like cells. Methods Combined approach of dexamethasone, HGF, IGF and other cytokines were used to induce the differentiation of hAECs into hepatocyte-like cells. The induction lasted 2 weeks. During the induction, the expression of albumin ALB, CYP1A1, CYP1A2, IGFR, c-met and key functional genes related to liver cells as well as transcription factors HNF3, HNF4 and C/EBPa were monitored by RT-PCR. Time dependent changes of the surface marker colony ALB, AFP and CK18 were analyzed by cell flow cytometry. Results After the 2 week induction, the expressions of liver hepatocyte-like cell functional genes such as albumin, CYP1A1, CYP1A2, c-met, and transcription factors such as HNF3, HNF4, C/EBPa and HNF1 were observed. Six days after the induction, hAECs mainly were stained AFP+, and the positive rate was (15.1±2.1)%. While 10 days after the induction, part of the hAECs showed AFP+/ALB+ (6.5±1.4)%; and on 14th day, hAECs only showed ALB+, and the rate was (13.9±2.3)%. ALB+ cell increase indicated a gradual functional maturation from the hAECs to hepatocyte-like cells. Similaritly, the number of CK18+ cells in the whole population was also increased: On 10th day, the rate was (16.1±1.2)%; on 14th day, that was (21.3±4.6)%, which proved the above hypothesis of the trandifferentiation. By extending the induction time, the expression of functional genes increased gradually, and a maturing process of hAECs was detected by cell surface markers. Conclusion The differentiation of hAECs induced in vitro has the characteristics of hepatocyte-like cells.
4.A case of severe acute pancreatitis and cholangitis presenting as initial complaints of hepatocellular carcinoma
Zuhai REN ; Hongwu LUO ; Yong XU ; Shaihong ZHU
Journal of Central South University(Medical Sciences) 2011;36(7):697-700
Jaundice occurs in 19%-40% of the hepatocellular carcinoma (HCC) patients. HCC associated jaundice may be divided into hepatocellular and icteric types in terms of its underlying pathophysiology. The jaundice of icteric type is caused by obstruction of the bile duct through cancer embolus, blood clot, biliary sludge, tumor compression or infiltration. Jaundice and epigastric discomforts are the main clinical manifestations. In the present case, severe acute pancreatitis and acute cholangitis presenting as initial complaints of icteric type HCC were quite rare. A tumor located at the central lobe of the liver and a cancer embolus at the lower part of the common bile duct (CBD) were detected by CT scan. Curative resection of HCC with CBD exploration eradicated both the tumor and the embolus, and no recurrence was found after a 36 month follow-up.
5.Argon plasma coagulation combined with bifurcated metal stents for treatment of airway stenosis in the carinal region A 16-case analysis
Hongwu WANG ; Yunzhi ZHOU ; Hongming MA ; Lingfei LUO ; Jing LI ; Hang ZOU ; Dongmei LI
Chinese Journal of Tissue Engineering Research 2009;13(30):5995-6000
A total of 16 patients with airway stenosis including benign lesion (n=4) and malignant disease (n=12) were treated with argon plasma coagulation (APC) and Z-type covered retrievable metallic stent.L-and I-type stents were placed by guidance of brenchoscope,while Y-type stent was placed by the guidance of both bronchoscope and fluoroscope.Airway stenosis was from (58.8 +9.1)% before APC to (7.5±2.4)% after APC in tracheal,from (67.4±7.4)% to (19.4±4.1)% in left main bronchus,from (69.6±8.9)% to (27.6±5.4)% in right main bronchus.Symptoms of chest distress and breathlessness were improved remarkably,tachypnea indexes were decreased,but Kamofsky performance scope were increased.Twelve stents were successfully installed by the first time,including 9 Y-shape;otherwise,4 stents failed to be installed because the carinal was too wide to insert the Y-shape stent in two patients with lung cancer,finally,L+I type stents were used.Median survival time after successful stenting was 10 months,and mean time was 13 months.APC combined with bifurcated metal stants relieved obstruction and improved quality of life.
6.Laparoscope and endoscope for portal hypertension.
Hongwu LUO ; Xiangjun HUANG ; Feizhou HUANG ; Xunyang LIU
Journal of Central South University(Medical Sciences) 2011;36(8):786-790
OBJECTIVE:
To determine the therapeutic effect of laparoscopic splenectomy, perisoph-agogastric devascularization, and endoscopic variceal ligation (EVL) on patients with portal hypertension.
METHODS:
We randomly divided 105 patients into 3 groups: 40 had endoscopic band ligation (the ligation group), 35 had splenectomy and perisoph-agogastric devascularization (the laparotomy group), and the other 30 had laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation (the combination group). Blood samples were analyzed preoperatively and postoperatively on day 1,3,and 7,including alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and directed bilirubin(DBIL). The length of stay, blood loss, operation time, anal exhaust time, azygos vein diameter, blood flow velocity and blood flow, recurrence of esophageal varices and rehaemorrhagia were compared.
RESULTS:
Between the combination group and the laparotomy group, the serum levels of TbIL and Dbil had difference on 1st postoperative day(P<0.05). AST had difference on 7th postoperative day(P<0.05). The length of stay, blood loss, operation time, and anal exhaust time had significant difference(P<0.05). Among the combination group, the laparotomy group and the ligation group, the azygos vein blood flow before and after the treatment, recurrence of esophageal varices and rehaemorrhagia had no difference(P<0.05).
CONCLUSION
Laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation have less trauma, lower recurrence rate, fewer complications and rapid recovery, and may reduce the azygous vein blood flow. It can be used safely for portal hypertension.
Adult
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Endoscopy
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methods
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Esophageal and Gastric Varices
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complications
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surgery
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Female
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Humans
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Hypertension, Portal
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complications
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surgery
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Laparoscopy
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methods
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Ligation
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methods
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Male
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Middle Aged
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Splenectomy
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methods
7.Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.
Hongwu, WANG ; Meifang, HAN ; Huaning, YAO ; Zhanhui, WANG ; Dong, XI ; Weiming, YAN ; Jinlin, HOU ; Xiaoping, LUO ; Qin, NING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(3):318-23
SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.
8.The inspiration of American pharmacists training system
Yue GAO ; Zhaoyuan LIU ; Li FAN ; Guoqiang DONG ; Zheng ZHOU ; Yungen LUO ; Hongwu ZHANG
Journal of Pharmaceutical Practice 2016;34(3):232-236
Taking Pharm .D .degree education in School of Pharmacy ,University of Pittsburgh as an example ,this pa-per introduced the American pharmacists training system ,degree setting for service-oriented pharmacy students ,training objec-tive ,course design ,teaching staff and other aspects .This paper also reviewed pharmacy education and professional develop-ments for licensed pharmacists in China .Based on the successful experience abroad ,proposals were made to modify the domes-tic pharmacy education .The process of personnel training should be emphasized to improve the quality and capability of licensed pharmacists in China .
9.Wolff's law-based continuum topology optimization method and its application in biomechanics.
Kun CAI ; Hongwu ZHANG ; Yangjun LUO ; Biaosong CHEN
Journal of Biomedical Engineering 2008;25(2):331-335
A new method for the simulation of the mass distribution of cancellous bone is presented on the basis of finite element analysis (FEA). In this method,the process of bone remodelling is considered as a process of the topology optimization of a corresponding continuum structure. Fabric tensor is used to express the microstructure and the constitutive properties of cancellous bone. The effective volume fraction or the relative density of a point in the design domain is expressed by the invariables of the fabric tensor. A reference strain interval, which is corresponding to the strain dead zone of a bone in biomechanics, is applied to detect the the final topology of the structure. By the present approach, several numerical results are given, i. e., the simulation on the shape of the coronal plane of vertebrae, the predictions of the mass distributions of the two-dimensional and the three-dimensional proximal femurs. The validity and feasibility of this new method are verified by the comparison between the results of the present work and those in the published literatures.
Adaptation, Physiological
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Algorithms
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Biomechanical Phenomena
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Bone Density
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physiology
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Bone Remodeling
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physiology
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Bone and Bones
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physiology
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Computer Simulation
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Femur Head
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physiology
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Finite Element Analysis
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Humans
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Models, Biological
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Stress, Mechanical
10.TACE with Ar-He cryosurgery combined minimal invasive technique for the treatment of primary NSCLC in 139 cases.
Lingfei LUO ; Hongwu WANG ; Hongming MA ; Hang ZOU ; Dongmei LI ; Yunzhi ZHOU
Chinese Journal of Lung Cancer 2010;13(1):60-63
BACKGROUND AND OBJECTIVETACE, Ar-He target cryosurgery and radioactive seeds implantation are the mainly micro-invasive methods in the treatment of lung cancer. This article summarizes the survival quality after treatment, the clinical efficiency and survival period, and analyzes the advantages and shortcomings of each methods so as to evaluate the clinical effect of non-small cell lung cancer with multiple minimally invasive treatment.
METHODSAll the 139 cases were non-small cell lung cancer patients confirmed by pathology and with follow up from July 2006 to July 2009 retrospectively, and all of them lost operative chance by comprehensive evaluation. Different combination of multiple minimally invasive treatments were selected according to the blood supply, size and location of the lesion. Among the 139 cases, 102 cases of primary and 37 cases of metastasis to mediastinum, lung and chest wall, 71 cases of abundant blood supply used the combination of superselective target artery chemotherapy, Ar-He target cryoablation and radiochemotherapy with seeds implantation; 48 cases of poor blood supply use single Ar-He target cryoablation; 20 cases of poor blood supply use the combination of Ar-He target cryoablation and radiochemotheraoy with seeds implantation. And then the pre- and post-treatment KPS score, imaging data and the result of follow up were analyzed.
RESULTSThe KPS score increased 20.01 meanly after the treatment. Follow up 3 years, 44 cases of CR, 87 cases of PR, 3 cases of NC and 5 cases of PD, and the efficiencywas 94.2%. Ninety-nine cases of 1 year survival (71.2%), 43 cases of 2 years survival (30.2%), 4 cases with over 3 years survival and the median survival was 19 months. Average survival was (16 +/- 1.5) months. There was no severe complications, such as spinal cord injury, vessel and pericardial aspiration.
CONCLUSIONMinimally invasive technique is a highly successful, micro-invasive and effective method with mild complications. To non-small cell lung cancer, we can improve the middle and long term clinical effect by using the different combination of multiple minimally invasive treatments according to the patient's condition.
Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Cryosurgery ; methods ; Female ; Humans ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome